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STEROID COMPOUND AND CONJUGATE THEREOF

20250002527 ยท 2025-01-02

    Inventors

    Cpc classification

    International classification

    Abstract

    The present invention relates to a steroid compound and a conjugate thereof, and specifically relates to a compound and a conjugate thereof, or a tautomer, a mesomer, a racemate, an enantiomer and a diastereoisomer thereof, or a mixture form thereof, or a pharmaceutically acceptable salt thereof; and a method for preparing the compound and the conjugate thereof and the use thereof.

    Claims

    1. A compound of formula I: ##STR01074## or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; B is absent or B is any group; W is absent or W is any group; A1 is a substituted benzene ring; CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; X is selected from the group consisting of: O, S and NR; Y.sub.1 is any group, and m is any integer from 0 to 4; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    2. The compound of formula I or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, selected from the group consisting of: ##STR01075## wherein, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; B is absent or B is any group; W is absent or W is any group; A1 is a substituted benzene ring; CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; X is selected from the group consisting of: O, S and NR; Y.sub.1 is any group, and m is 0 or 1; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    3. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-2, wherein X is O, S or NH.

    4. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S and C(O).

    5. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    6. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 5, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl, or R.sub.4 and R.sub.5 together form C.sub.3-C.sub.6 cycloalkyl or 3-6 membered heterocyclyl; and n is selected from the group consisting of 1, 2 and 3.

    7. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 6, wherein the (CR.sub.4R.sub.5).sub.n are each independently selected from the group consisting of CH.sub.2, ##STR01076## CH.sub.2CH.sub.2 and ##STR01077##

    8. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein Y.sub.1 is absent or selected from the group consisting of: protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    9. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-8, wherein Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and optionally substituted C.sub.1-C.sub.6 alkyl.

    10. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-8, wherein A1 is selected from the group consisting of: ##STR01078## ##STR01079##

    11. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-10, wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted C.sub.1-C.sub.6 alkyl.

    12. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-11, wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    13. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-12, wherein R.sub.3 is selected from the group consisting of: hydrogen, optionally substituted OH, optionally substituted SH and optionally substituted C.sub.1-C.sub.6 alkyl.

    14. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-13, wherein R.sub.3 is selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted ##STR01080## optionally substituted ##STR01081## optionally substituted ##STR01082## optionally substituted ##STR01083## optionally substituted ##STR01084## optionally substituted ##STR01085## optionally substituted ##STR01086## optionally substituted ##STR01087## optionally substituted ##STR01088## optionally substituted ##STR01089## optionally substituted ##STR01090## optionally substituted ##STR01091## optionally substituted ##STR01092## optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted-OCH.sub.2F, optionally substituted-OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    15. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-14, wherein R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH, ##STR01093## OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    16. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-15, wherein B is selected from the group consisting of: optionally substituted ##STR01094## optionally substituted ##STR01095## and optionally substituted ##STR01096## and X.sub.1 is selected from the group consisting of: CH, C(OCH.sub.3) and N.

    17. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-15, wherein B is selected from the group consisting of: optionally substituted ##STR01097## optionally substituted ##STR01098## optionally substituted ##STR01099## optionally substituted ##STR01100## optionally substituted ##STR01101## optionally substituted ##STR01102## optionally substituted ##STR01103## optionally substituted ##STR01104## optionally substituted ##STR01105## optionally substituted ##STR01106## optionally substituted ##STR01107## optionally substituted ##STR01108## optionally substituted ##STR01109## optionally substituted ##STR01110## optionally substituted ##STR01111## optionally substituted ##STR01112## optionally substituted ##STR01113## optionally substituted ##STR01114## optionally substituted ##STR01115## and optionally substituted ##STR01116## wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    18. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-17, wherein B is selected from the group consisting of: ##STR01117##

    19. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-18, wherein W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted CH.sub.2NHC(O), optionally substituted NHC(O)CH.sub.2, optionally substituted C(O)NH, optionally substituted NH, optionally substituted CHCH, CC, optionally substituted C.sub.1-C.sub.6 alkylene, optionally substituted OCH.sub.2, optionally substituted CH.sub.2O, optionally substituted SCH.sub.2, optionally substituted CH.sub.2S, optionally substituted NHC(O), optionally substituted C(O)CH.sub.2, optionally substituted CH.sub.2NH, optionally substituted NHCH.sub.2, optionally substituted ##STR01118## optionally substituted ##STR01119## optionally substituted ##STR01120## optionally substituted ##STR01121## and optionally substituted ##STR01122##

    20. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 19, wherein W is absent or W is selected from the group consisting of: ##STR01123##

    21. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 20, wherein W is absent or W is ##STR01124##

    22. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-21, wherein B is absent or selected from the group consisting of: ##STR01125## W is absent or W is selected from the group consisting of: ##STR01126## A1 is selected from the group consisting of: ##STR01127##

    23. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-22, selected from the group consisting of ##STR01128## wherein, X is selected from the group consisting of O, S and NH; R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6alkyl; n is selected from the group consisting of 1, 2 and 3; Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH, ##STR01129## OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    24. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-23, wherein the compound is selected from the group consisting of the following structures: TABLE-US-00028 No. Structure I-1 embedded image I-2 embedded image I-3 embedded image I-4 embedded image I-5 embedded image I-6 embedded image I-7 embedded image I-8 embedded image I-9 embedded image I-10 embedded image I-11 embedded image I-12 embedded image I-13 embedded image I-14 embedded image I-15 embedded image I-16 embedded image I-17 embedded image I-18 embedded image I-19 embedded image I-20 embedded image I-21 embedded image I-22 embedded image I-23 embedded image I-24 embedded image I-25 embedded image I-26 embedded image I-27 embedded image I-28 embedded image I-29 embedded image I-30 embedded image I-31 embedded image I-32 embedded image I-33 embedded image I-34 embedded image I-35 embedded image I-36 embedded image I-37 embedded image I-38 embedded image I-39 embedded image I-40 embedded image I-41 embedded image I-42 embedded image I-43 embedded image I-44 embedded image I-45 embedded image I-46 embedded image I-47 embedded image I-48 embedded image I-49 embedded image I-50 embedded image I-51 embedded image I-52 embedded image I-53 embedded image I-54 embedded image I-55 embedded image I-56 embedded image I-57 embedded image I-58 embedded image I-59 embedded image I-60 embedded image I-61 embedded image I-62 embedded image I-63 embedded image I-64 embedded image I-65 embedded image I-66 embedded image I-67 embedded image I-68 embedded image I-69 embedded image I-70 embedded image I-71 embedded image I-72 embedded image I-73 embedded image I-74 embedded image I-75 embedded image I-76 embedded image I-77 embedded image I-78 embedded image I-79 embedded image I-80 embedded image I-81 embedded image I-82 embedded image I-83 embedded image I-84 embedded image I-85 embedded image I-86 embedded image I-87 embedded image I-88 embedded image I-89 embedded image I-90 embedded image I-91 embedded image I-92 embedded image I-93 embedded image I-94 embedded image I-95 embedded image I-96 embedded image I-97 embedded image I-98 embedded image I-99 embedded image I-100 embedded image I-101 embedded image I-102 embedded image I-103 embedded image I-104 embedded image I-105 embedded image I-106 embedded image I-107 embedded image I-108 embedded image I-109 embedded image I-110 embedded image I-111 embedded image I-112 embedded image I-113 embedded image I-114 embedded image I-115 embedded image I-116 embedded image I-117 embedded image I-118 embedded image I-119 embedded image I-120 embedded image I-121 embedded image I-122 embedded image I-123 embedded image I-124 embedded image I-125 embedded image I-126 embedded image I-127 embedded image I-128 embedded image I-129 embedded image I-130 embedded image I-131 embedded image I-132 embedded image I-133 embedded image I-134 embedded image I-135 embedded image I-136 embedded image I-137 embedded image I-138 embedded image I-139 embedded image I-140 embedded image I-141 embedded image I-142 embedded image I-143 embedded image I-144 embedded image I-145 embedded image I-146 embedded image I-147 embedded image I-148 embedded image I-149 embedded image I-150 embedded image I-151 embedded image I-152 embedded image I-153 embedded image I-154 embedded image I-155 embedded image I-156 embedded image I-157 embedded image I-158 embedded image I-159 embedded image I-160 embedded image I-161 embedded image I-162 embedded image I-163 embedded image I-164 embedded image I-165 embedded image I-166 embedded image I-167 embedded image I-168 embedded image I-169 embedded image I-170 embedded image I-171 embedded image I-172 embedded image I-173 embedded image I-174 embedded image I-175 embedded image I-176 embedded image I-177 embedded image I-178 embedded image I-179 embedded image I-180 embedded image I-181 embedded image I-182 embedded image I-183 embedded image I-184 embedded image I-185 embedded image I-186 embedded image I-187 embedded image I-188 embedded image I-189 embedded image I-190 embedded image I-191 embedded image I-192 embedded image I-193 embedded image I-194 embedded image I-195 embedded image I-196 embedded image I-197 embedded image I-198 embedded image I-199 embedded image I-200 embedded image I-201 embedded image I-202 embedded image I-203 embedded image I-204 embedded image I-205 embedded image I-206 embedded image I-207 embedded image I-208 embedded image I-209 embedded image I-210 embedded image I-211 embedded image I-212 embedded image I-213 embedded image I-214 embedded image I-215 embedded image I-216 embedded image I-217 embedded image I-218 embedded image I-219 embedded image I-220 embedded image I-221 embedded image I-222 embedded image I-223 embedded image I-224 embedded image I-225 embedded image I-226 embedded image I-227 embedded image I-228 embedded image I-229 embedded image I-230 embedded image I-231 embedded image I-232 embedded image I-233 embedded image I-234 embedded image I-235 embedded image I-236 embedded image I-237 embedded image I-238 embedded image I-239 embedded image I-240 embedded image I-241 embedded image I-242 embedded image I-243 embedded image I-244 embedded image I-245 embedded image I-246 embedded image I-247 embedded image I-248 embedded image I-249 embedded image I-250 embedded image I-251 embedded image I-252 embedded image I-253 embedded image I-254 embedded image I-255 embedded image I-256 embedded image I-257 embedded image I-258 embedded image I-259 embedded image I-260 embedded image I-261 embedded image I-262 embedded image I-263 embedded image I-264 embedded image I-265 embedded image I-266 embedded image I-267 embedded image I-268 embedded image I-269 embedded image I-270 embedded image I-271 embedded image I-272 embedded image I-273 embedded image I-274 embedded image I-275 embedded image I-276 embedded image I-277 embedded image I-278 embedded image I-279 embedded image I-280 embedded image I-281 embedded image I-282 embedded image I-283 embedded image I-284 embedded image I-285 embedded image I-286 embedded image I-287 embedded image I-288 embedded image I-289 embedded image I-290 embedded image I-291 embedded image I-292 embedded image I-293 embedded image I-294 embedded image I-295 embedded image I-296 embedded image I-297 embedded image I-298 embedded image I-299 embedded image I-300 embedded image I-301 embedded image I-302 embedded image I-303 embedded image I-304 embedded image I-305 embedded image I-306 embedded image I-307 embedded image I-308 embedded image I-309 embedded image I-310 embedded image I-311 embedded image I-312 embedded image I-313 embedded image I-314 embedded image I-315 embedded image I-316 embedded image I-317 embedded image I-318 embedded image I-319 embedded image I-320 embedded image I-321 embedded image I-322 embedded image I-323 embedded image I-324 embedded image I-325 embedded image I-326 embedded image I-327 embedded image I-328 embedded image I-329 embedded image I-330 embedded image I-331 embedded image I-332 embedded image I-333 embedded image I-334 embedded image I-335 embedded image I-336 embedded image I-337 embedded image I-338 embedded image I-339 embedded image I-340 embedded image I-341 embedded image I-342 embedded image I-343 embedded image I-344 embedded image I-345 embedded image I-346 embedded image I-347 embedded image I-348 embedded image I-349 embedded image I-350 embedded image I-351 embedded image I-352 embedded image I-353 embedded image I-354 embedded image I-355 embedded image I-356 embedded image I-357 embedded image I-358 embedded image I-359 embedded image I-360 embedded image I-361 embedded image I-362 embedded image I-363 embedded image

    25. A compound of a formula below or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: ##STR01493##

    26. A compound of formula IIa or IIb: ##STR01494## or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; B is absent or B is any group; W is absent or W is any group; A2 is a substituted benzene ring; CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; X is selected from the group consisting of: O, S and NR; Y.sub.1 is any group, and m is any integer from 0 to 4; Y.sub.2 is selected from the group consisting of O, S and NR; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; the wavy line custom-character in the general formula represents being directly linked to a ligand via the X or Y.sub.2 group, or being linked to the ligand via a linker fragment.

    27. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 26, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    28. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 27, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl; and n is selected from the group consisting of 1, 2 and 3.

    29. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-28, wherein Y.sub.1 is absent or selected from the group consisting of: OR, SR, N(R).sub.2 and optionally substituted C.sub.1-C.sub.6 alkyl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy.

    30. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 29, wherein Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl.

    31. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-30, wherein A2 is selected from the group consisting of: ##STR01495## ##STR01496## ##STR01497##

    32. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-31, wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted C.sub.1-C.sub.6 alkyl.

    33. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-33, wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    34. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 58-71, wherein R.sub.3 is selected from the group consisting of: hydrogen, optionally substituted OH, optionally substituted SH and optionally substituted C.sub.1-C.sub.6 alkyl.

    35. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-34, wherein R.sub.3 is selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted ##STR01498## optionally substituted ##STR01499## optionally substituted ##STR01500## optionally substituted ##STR01501## optionally substituted ##STR01502## optionally substituted ##STR01503## optionally substituted ##STR01504## optionally substituted ##STR01505## optionally substituted ##STR01506## optionally substituted ##STR01507## optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted OCH.sub.2F, optionally substituted OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    36. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-35, wherein R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH, ##STR01508## OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    37. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-36, wherein B is selected from the group consisting of: optionally substituted ##STR01509## optionally substituted ##STR01510## and optionally substituted ##STR01511## and X.sub.1 is selected from the group consisting of: CH, C(OCH.sub.3) and N.

    38. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-36, wherein B is selected from the group consisting of: optionally substituted ##STR01512## optionally substituted ##STR01513## optionally substituted ##STR01514## optionally substituted ##STR01515## optionally substituted ##STR01516## optionally substituted ##STR01517## optionally substituted ##STR01518## optionally substituted ##STR01519## optionally substituted ##STR01520## optionally substituted ##STR01521## optionally substituted ##STR01522## optionally substituted ##STR01523## optionally substituted ##STR01524## optionally substituted ##STR01525## optionally substituted ##STR01526## optionally substituted ##STR01527## optionally substituted ##STR01528## optionally substituted ##STR01529## optionally substituted ##STR01530## and optionally substituted ##STR01531## wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    39. The compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-38, wherein B is selected from the group consisting of: ##STR01532##

    40. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-39, wherein W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted CH.sub.2NHC(O), optionally substituted NHC(O)CH.sub.2, optionally substituted C(O)NH, optionally substituted NH, optionally substituted CHCH, CC, optionally substituted C.sub.1-C.sub.6 alkylene, optionally substituted OCH.sub.2, optionally substituted CH.sub.2O, optionally substituted SCH.sub.2, optionally substituted CH.sub.2S, optionally substituted NHC(O), optionally substituted COCH.sub.2, optionally substituted CH.sub.2NH, optionally substituted NHCH.sub.2, optionally substituted CH(CH.sub.3), optionally substituted ##STR01533## optionally substituted ##STR01534## optionally substituted ##STR01535## optionally substituted ##STR01536## and optionally substituted ##STR01537##

    41. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 40, wherein W is absent or W is selected from the group consisting of: ##STR01538##

    42. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 41, wherein W is absent or W is ##STR01539##

    43. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-42, wherein B is absent or selected from the group consisting of: ##STR01540## W is absent or W is selected from the group consisting of: ##STR01541## A2 is selected from the group consisting of: ##STR01542## ##STR01543## ##STR01544## ##STR01545##

    44. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-43, selected from the group consisting of: ##STR01546## wherein, X is selected from the group consisting of O, S and NH; R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6alkyl; n is selected from the group consisting of 1, 2 and 3; Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH, ##STR01547## OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    45. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-44, wherein the compound is selected from the group consisting of: ##STR01548## ##STR01549##

    46. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-45, wherein the compound further comprises a linker fragment, and the compound of formula IIa or IIb is capable of being coupled to a ligand via the linker fragment, wherein the linker fragment comprises a L.sub.1 fragment, a L.sub.2 fragment and/or an L.sub.3 fragment, and the compound has the following structure: ##STR01550## wherein, Tr is absent or Tr is any group; L.sub.3 is selected from a polypeptide fragment; L.sub.2 is absent or selected from a linker fragment; L.sub.1 is selected from a coupling unit; R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; B is absent or B is any group; W is absent or W is any group; CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; X is selected from the group consisting of: O, S and NR; Y.sub.1 is any group, and m is any integer from 0 to 4; Y.sub.2 is selected from the group consisting of O, S and NR; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    47. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 46, wherein Tr is absent or selected from the group consisting of the following structures: ##STR01551##

    48. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 47, wherein L.sub.3 is selected from the group consisting of a dipeptide, a tripeptide, and a tetrapeptide, wherein the dipeptide is selected from the group consisting of: GA, GG, AG, EG, EA, GE, DG, DA, GD, VC, VA, AA and VK, the tripeptide is selected from the group consisting of: EAG, EGG, GEG, GEA, DAG, DGG, GDG, GDA, GGA, GAG, GFG, AAG, AAA, VAG, VCG and VKG, and the tetrapeptide is selected from the group consisting of: GGFG, GGAG, GGGG, GEGG, GEAG, GDGG, GDAG, AAAG and EAGG.

    49. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 48, wherein L.sub.3 is selected from the group consisting of: glycine-glycine-phenylalanine-glycine (GGFG), alanine-alanine-alanine-glycine (AAAG), glycine-glycine-glycine-glycine (GGGG), valine-alanine-glycine (VAG), valine-citrulline-glycine (VCG), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine-alanine (AA), glutamic acid-alanine-glycine-glycine (EAGG), glycine-glutamic acid-alanine-glycine (GEAG), glycine-glutamic acid-glycine-glycine (GEGG), glutamic acid-glycine-glycine (EGG), glutamic acid-alanine-glycine (EAG), valine-lysine-glycine (VKG), glycine-glutamic acid-glycine (GEG), glutamic acid-alanine (EA), glutamic acid-glycine (EG) and glycine-glutamic acid (GE).

    50. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 46-49, wherein L.sub.2 is absent, or L.sub.2 comprises or does not comprise a PEG branch chain or a PEG linear chain.

    51. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 50, wherein (1) when L.sub.2 does not comprise PEG, L.sub.2 is selected from the group consisting of: ##STR01552## (2) when L.sub.2 comprises the PEG linear chain, L.sub.2 is selected from the group consisting of: ##STR01553## wherein p is ay integer from 1 to 20; (3) when L.sub.2 comprises the PEG branch chain, L.sub.2 is selected from the group consisting of: ##STR01554## ##STR01555## ##STR01556## ##STR01557## wherein q is selected from any integer from 1 to 30.

    52. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 51, wherein (1) when L.sub.2 comprises the PEG linear chain, L.sub.2 is selected from the group consisting of: ##STR01558## (2) when L.sub.2 comprises the PEG branch chain, L.sub.2 is selected from the group consisting of: ##STR01559## ##STR01560## ##STR01561## ##STR01562## ##STR01563## ##STR01564## ##STR01565## ##STR01566##

    53. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 46-52, wherein, (1) when L.sub.1 is coupled to the ligand via sulfhydryl, L.sub.1 is selected from the group consisting of: ##STR01567## ##STR01568## wherein, R.sup.L1a, R.sup.L1b and R.sup.L1c are each independently selected from the group consisting of: hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted aryl and optionally substituted benzyl; (2) when L.sub.1 is coupled to the ligand via amino, L.sub.1 is selected from the group consisting of: ##STR01569## (3) when L.sub.1 is coupled via click chemistry, L.sub.1 is selected from the group consisting of: ##STR01570##

    54. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 46-53, wherein the compound of formula IVa or IVb is selected from the group consisting of the following structures: TABLE-US-00029 No. Structure IV-1 embedded image IV-2 embedded image IV-6 embedded image IV-7 embedded image IV-11 embedded image IV-12 embedded image IV-13 embedded image IV-14 embedded image IV-15 embedded image IV-16 embedded image IV-17 embedded image IV-21 embedded image IV-25 embedded image IV-29 embedded image IV-30 embedded image IV-34 embedded image IV-35 embedded image IV-39 embedded image IV-43 embedded image IV-44 embedded image IV-45 embedded image IV-46 embedded image IV-47 embedded image IV-48 embedded image IV-49 embedded image IV-50 embedded image IV-51 embedded image IV-52 embedded image IV-53 embedded image IV-54 embedded image IV-55 embedded image IV-56 embedded image IV-57 embedded image IV-58 embedded image IV-59 embedded image IV-60 embedded image IV-61 embedded image IV-62 embedded image IV-63 embedded image IV-64 embedded image IV-65 embedded image IV-66 embedded image IV-67 embedded image IV-68 embedded image IV-69 embedded image IV-70 embedded image IV-71 embedded image IV-81 embedded image IV-82 embedded image IV-83 embedded image IV-84 embedded image IV-91 embedded image IV-92 embedded image IV-93 embedded image IV-94 embedded image IV-101 embedded image IV-102 embedded image IV-103 embedded image IV-104 embedded image IV-111 embedded image IV-112 embedded image IV-113 embedded image IV-114 embedded image IV-115 embedded image IV-116 embedded image IV-117 embedded image IV-118 embedded image IV-119 embedded image IV-120 embedded image IV-121 embedded image IV-122 embedded image IV-123 embedded image IV-124 embedded image IV-125 embedded image IV-126 embedded image IV-127 embedded image IV-128 embedded image IV-129 embedded image IV-130 embedded image IV-131 embedded image IV-141 embedded image IV-142 embedded image IV-143 embedded image IV-144 embedded image IV-145 embedded image IV-146 embedded image IV-147 embedded image IV-148 embedded image IV-149 embedded image IV-150 embedded image IV-151 embedded image IV-152 embedded image IV-153 embedded image IV-154 embedded image IV-155 embedded image IV-156 embedded image IV-157 embedded image IV-158 embedded image IV-159 embedded image IV-160 embedded image

    55. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 54, wherein the compound of formula IVa or IVb is selected from the group consisting of the following structures: ##STR01671## ##STR01672##

    56. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 26-45, wherein the compound further comprises a linker fragment, the compound of formula IIa or IIb is capable of being coupled to the ligand via the linker fragment, and the compound has the following structure: ##STR01673## wherein, Tr is absent or Tr is any group; L.sub.3 is selected from a polypeptide fragment; L.sub.2 is absent or selected from a linker fragment; L.sub.1 is selected from a coupling unit; and L.sub.1 in the formulas IVa-1 and IVb-1 is in a linked form; R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, B, W, CR.sub.4R.sub.5, n, X, Y.sub.1 and Y.sub.2 are each as described in any one of claims 26-45; the wavy line custom-character in the general formulas represents being linked to the ligand via the L.sub.1 group.

    57. The compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 56, wherein the structural unit -Tr-L.sub.3-L.sub.2-L.sub.1- is selected from the group consisting of: ##STR01674##

    58. A conjugate, comprising the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-57, wherein the conjugate comprises a ligand-drug conjugate.

    59. The conjugate according to claim 58, wherein the ligand comprises an antibody or an antigen-binding fragment thereof.

    60. The conjugate according to claim 59, wherein the antibody is selected from the group consisting of: a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, a monoclonal antibody and a polyclonal antibody; the antigen-binding fragment is selected from the group consisting of: a Fab, a Fab, a F(ab)2, a Fv, a scFv, a diabody, a Fd, a dAb, a VHH, a maxibody and a complementarity determining region (CDR) fragment.

    61. The conjugate according to claims 58-60, wherein the ligand specifically binds to an antigen selected from the group consisting of: AXL, BAFFR, BCMA, BCR-list components, BDCA2, BDCA4, BTLA, BTNL2 BTNL3, BTNL8, BTNL9, C.sub.10orf54, CCR1, CCR3, CCR4, CCR5, CCR6, CCR7, CCR9, CCR10, CD11c, CD137, CD138, CD14, CD163, CD168, CD177, CD19, CD20, CD209, CD209L, CD22, CD226, CD248, CD25, CD27, CD274, CD276, CD28, CD30, CD300A, CD33, CD37, CD38, CD4, cluster of differentiation 40 (CD40), CD44, CD45, CD46, CD47, CD48, CD5, CD52, CD55, CD56, CD59, CD62E, CD68, CD69, CD70, CD74, CD79a, CD79b, CD8, CD80, CD86, CD90.2, CD96, CLEC12A, CLEC12B, CLEC7A, CLEC9A, CR1, CR3, CRTAM, CSF1R, CTLA4, CXCR1/2, CXCR4, CXCR5, DDR1, DDR2, DEC-205, DLL4, DR6, FAP, FCamR, FCMR, FcR's, Fire, GITR, HHLA2, HLA class II, HVEM, ICOSLG, IFNAR, type I interferon receptor subunit (IFNAR1), IFNLR1, IL10R.sub.1, IL10R.sub.2, IL12R, IL13RA1, IL13RA2, IL15R, IL17RA, IL17RB, IL17RC, IL17RE, IL20R.sub.1, IL20R.sub.2, IL21R, IL22R.sub.1, IL22RA, IL23R, IL27R, IL29R, IL2Rg, IL31R, IL36R, IL3RA, IL4R, IL6R, IL5R, IL7R, IL9R, integrins, LAG3, LIFR, MAG/Siglec-4 (sialic acid-binding immunoglobulin-like lectin-4), MMR, MSR1, NCR3LG1, NKG2D, NKp30, NKp46, OX40 (CD134), PDCD1, PROKR1, PVR, PVRIG, PVRL2, PVRL3, RELT, SIGIRR, Siglec-1 (sialic acid-binding immunoglobulin-like lectin-1), Siglec-10, Siglec-5, Siglec-6, Siglec-7, Siglec-8, Siglec-9, SIRPA, SLAMF7, TACI, TCR-list components/assoc, PTCRA, TCRb, CD3z, CD3, TEK, TGFBR1, TGFBR2, TGFBR3, TIGIT, TLR2, TLR4, tumor necrosis factor (TNF), TROY, TSLPR, TYRO, VLDLR, VSIG4, IL2R-y and VTCN1.

    62. The conjugate according to claim 61, wherein the ligand is selected from the group consisting of: an anti-TNF antibody or an antigen-binding fragment thereof, an anti-CD40 antibody or an antigen-binding fragment thereof, and an anti-IFNAR1 antibody or an antigen-binding fragment thereof.

    63. The conjugate according to claims 58-62, wherein the ligand is selected from the group consisting of: adalimumab, iscalimab (CFZ533), anifrolumab (MEDI-546), infliximab, afelimomab, golimumab, BIIB059, 8c11, and derivatives and biosimilars thereof.

    64. The conjugate according to claims 58-63, wherein the ligand-drug conjugate has the following structure: ##STR01675## wherein, Ab represents a ligand capable of binding to a target, and N.sup.a-I is any number from 1 to 10; Tr is absent or Tr is any group; L.sub.3 is selected from a polypeptide fragment; L.sub.2 is absent or selected from a linker fragment; L.sub.1 is selected from a coupling unit; R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; B is absent or B is any group; W is absent or W is any group; CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; X is selected from the group consisting of: O, S and NR; Y.sub.1 is any group, and m is any integer from 0 to 4; Y.sub.2 is selected from the group consisting of O, S and NR; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    65. The conjugate according to claim 64, wherein when L.sub.1 is coupled to Ab via sulfhydryl, L.sub.1 is selected from the group consisting of the following structures: ##STR01676## ##STR01677## wherein R.sup.L1c is selected from the group consisting of: hydrogen, optionally substituted alkyl and optionally substituted aryl; when L.sub.1 is coupled to Ab via amino, L.sub.1 is selected from the group consisting of the following structures: ##STR01678## when L.sub.1 is coupled to Ab via click chemistry, L.sub.1 is selected from the group consisting of the following structures: ##STR01679##

    66. The conjugate according to any one of claims 59-65, wherein the conjugate is selected from the group consisting of the following structures: TABLE-US-00030 No. Structure V-1 embedded image V-2 embedded image V-6 embedded image V-7 embedded image V-8 embedded image V-12 embedded image V-13 embedded image V-14 embedded image V-15 embedded image V-16 embedded image V-17 embedded image V-18 embedded image V-22 embedded image V-25 embedded image V-29 embedded image V-30 embedded image V-34 embedded image V-35 embedded image V-39 embedded image V-43 embedded image V-44 embedded image V-45 embedded image V-46 embedded image V-47 embedded image V-48 embedded image V-49 embedded image V-50 embedded image V-51 embedded image V-52 embedded image V-53 embedded image V-54 embedded image V-55 embedded image V-56 embedded image V-57 embedded image V-58 embedded image V-59 embedded image V-60 embedded image V-70 embedded image V-74 embedded image V-75 embedded image V-78 embedded image V-81 embedded image V-84 embedded image V-87 embedded image V-90 embedded image V-91 embedded image V-92 embedded image V-95 embedded image V-98 embedded image V-102 embedded image V-103 embedded image V-104 embedded image V-105 embedded image V-106 embedded image V-109 embedded image V-110 embedded image V-111 embedded image V-112 embedded image V-113 embedded image V-114 embedded image V-120 embedded image V-121 embedded image V-122 embedded image V-123 embedded image V-124 embedded image V-125 embedded image V-126 embedded image V-127 embedded image V-128 embedded image V-129 embedded image V-130 embedded image V-131 embedded image V-132 embedded image V-133 embedded image V-134 embedded image V-135 embedded image V-136 embedded image V-137 embedded image V-138 embedded image V-139 embedded image V-140 embedded image V-141 embedded image V-145 embedded image V-146 embedded image V-147 embedded image V-151 embedded image V-152 embedded image V-153 embedded image V-154 embedded image V-155 embedded image V-156 embedded image V-157 embedded image V-161 embedded image V-164 embedded image V-168 embedded image V-169 embedded image V-173 embedded image V-174 embedded image V-178 embedded image V-182 embedded image V-183 embedded image V-184 embedded image V-185 embedded image V-186 embedded image V-187 embedded image V-188 embedded image V-189 embedded image V-190 embedded image V-191 embedded image V-192 embedded image V-193 embedded image V-194 embedded image V-195 embedded image V-196 embedded image V-197 embedded image V-198 embedded image V-199 embedded image V-209 embedded image V-213 embedded image V-216 embedded image V-219 embedded image V-222 embedded image V-225 embedded image V-228 embedded image V-229 embedded image V-230 embedded image V-233 embedded image V-236 embedded image V-240 embedded image V-241 embedded image V-242 embedded image V-243 embedded image V-244 embedded image V-247 embedded image V-248 embedded image V-249 embedded image V-250 embedded image V-251 embedded image V-252 embedded image V-258 embedded image V-259 embedded image V-260 embedded image V-261 embedded image V-262 embedded image V-263 embedded image V-264 embedded image V-265 embedded image V-266 embedded image V-267 embedded image V-268 embedded image V-269 embedded image V-270 embedded image V-271 embedded image V-272 embedded image V-273 embedded image V-274 embedded image V-275 embedded image V-276 embedded image V-277 embedded image V-278 embedded image V-279 embedded image V-283 embedded image V-284 embedded image V-285 embedded image V-289 embedded image V-290 embedded image V-291 embedded image V-292 embedded image V-293 embedded image V-294 embedded image V-295 embedded image V-299 embedded image V-302 embedded image V-306 embedded image V-307 embedded image V-311 embedded image V-312 embedded image V-316 embedded image V-320 embedded image V-321 embedded image V-322 embedded image V-323 embedded image V-324 embedded image V-325 embedded image V-326 embedded image V-327 embedded image V-328 embedded image V-329 embedded image V-330 embedded image V-331 embedded image V-332 embedded image V-333 embedded image V-334 embedded image V-335 embedded image V-336 embedded image V-337 embedded image V-347 embedded image V-351 embedded image V-354 embedded image V-357 embedded image V-360 embedded image V-363 embedded image V-366 embedded image V-367 embedded image V-368 embedded image V-371 embedded image V-374 embedded image V-378 embedded image V-379 embedded image V-380 embedded image V-381 embedded image V-382 embedded image V-385 embedded image V-386 embedded image V-387 embedded image V-388 embedded image V-389 embedded image V-390 embedded image V-396 embedded image V-397 embedded image V-398 embedded image V-399 embedded image V-400 embedded image V-401 embedded image V-402 embedded image V-403 embedded image V-404 embedded image V-405 embedded image V-406 embedded image V-407 embedded image V-408 embedded image V-409 embedded image V-410 embedded image V-411 embedded image V-412 embedded image V-413 embedded image V-414 embedded image V-415 embedded image wherein N.sup.a-I is any number from 1 to 10.

    67. The conjugate according to claims 58-65, wherein the ligand-drug conjugate has the following structure: ##STR01918## wherein, Ab represents a ligand capable of binding to a target, and N.sup.a-I is any number from 1 to 10; X is selected from the group consisting of O, S and NH; R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl; n is selected from the group consisting of 1, 2 and 3; Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH, ##STR01919## OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    68. The conjugate according to claims 58-65, wherein the conjugate is selected from the group consisting of the following structures: ##STR01920## ##STR01921## wherein N.sup.a-I is any number from 1 to 10, and Ab is selected from an antibody or an antigen-binding fragment thereof.

    69. A ligand-drug conjugate of a formula below or a tautomer, an enantiomer or a diastereoisomer thereof, or a mixture of isomers thereof, or a pharmaceutically acceptable salt or a solvate thereof, wherein the ligand-drug conjugate is selected from the group consisting of the following structures: ##STR01922## ##STR01923## ##STR01924## ##STR01925## wherein N.sup.a-I is any number from 1 to 10.

    70. A pharmaceutical composition, comprising the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-57 and/or the conjugate according to any one of claims 58-68, and optionally a pharmaceutically acceptable carrier.

    71. A method for influencing immune system functions, comprising administering to a subject the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-57, the conjugate according to any one of claims 58-68, and/or the pharmaceutical composition according to claim 69.

    72. Use of the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof according to any one of claims 1-57, the conjugate according to any one of claims 58-68 and/or the pharmaceutical composition according to claim 69 in the preparation of a medicament for preventing and/or treating diseases and/or conditions, wherein the diseases and/or the conditions include diseases and/or conditions associated with glucocorticoid receptor signaling.

    73. The use according to claim 72, wherein the diseases and/or the conditions are selected from the group consisting of: rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjogren's syndrome, ankylosing spondylitis, Wegener's granulomatosis and systemic sclerosis, autoimmune hemolytic anemia, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia and vasculitis, multiple sclerosis, myasthenia gravis and Guillain-Barre syndrome, ulcerative colitis, Crohn's disease, autoimmune diseases and atrophic gastritis, IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, Goodpasture's syndrome and lupus nephritis, type I diabetes, Grave's disease, Hashimoto's thyroiditis, primary adrenocortical atrophy and chronic thyroiditis, psoriasis, pemphigus vulgaris, cutaneous lupus erythematosus, dermatomyositis and polymyalgia rheumatica, and asthma.

    Description

    BRIEF DESCRIPTION OF THE DRAWINGS

    [0094] FIG. 1 shows the inhibiting effect of the drug conjugate of the present invention on the R848-induced release of IFN, TNF, IL-6 and IL-8 from human peripheral blood mononuclear cells. 1a: R848-induced IFN release from PBMCs; 1b: R848-induced TNF release from PBMCs; 1c: R848-induced IL6 release from PBMCs, 1d: R848-induced IL8 release from PBMCs.

    [0095] FIG. 2 shows the determination of biological activity in fluorescein isothiocyanate (FITC)-induced delayed-type IV hypersensitivity mouse models.

    [0096] FIG. 3 shows the arthritis scores for bovine type II collagen mixed adjuvant-induced DBA/1 mouse arthritis models. 3a: arthritis scores; 3b: AUC for arthritis scores.

    DETAILED DESCRIPTION OF THE INVENTION

    ##STR00005##

    or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein, [0097] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0098] B is absent or B is any group; [0099] W is absent or W is any group; [0100] A1 is a substituted benzene ring; [0101] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0102] X is selected from the group consisting of: O, S and NR; [0103] Y.sub.1 is any group, and m is any integer from 0 to 4; [0104] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0105] In certain embodiments, the compound of formula I or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof is selected from the group consisting of:

    ##STR00006## [0106] wherein, [0107] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0108] B is absent or B is any group; [0109] W is absent or W is any group; [0110] A1 is a substituted benzene ring; [0111] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0112] X is selected from the group consisting of: O, S and NR; [0113] Y.sub.1 is any group, and m is 0 or 1; [0114] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0115] In certain preferred embodiments of the present invention, certain groups in the compounds of formulas (I), (Ia), (Ib) and (Ic) or pharmaceutically acceptable salts thereof are defined as follows, and groups not mentioned are as described in any one of the embodiments of the present application (in certain embodiments for short).

    [0116] In certain embodiments, X is O, S or NH.

    [0117] In certain embodiments, CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S and C(O).

    [0118] In certain embodiments, R.sub.4 and R.sub.5 together form optionally substituted cycloalkyl or optionally substituted heterocyclyl.

    [0119] In certain embodiments, R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0120] In certain embodiments, R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0121] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl, or R.sub.4 and R.sub.5 together form C.sub.3-C.sub.6 cycloalkyl or 3-6 membered heterocyclyl; and n is selected from the group consisting of 1, 2 and 3.

    [0122] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein the (CR.sub.4R.sub.5).sub.n are each independently selected from the group consisting of CH.sub.2, CH.sub.2CH.sub.2

    ##STR00007##

    [0123] In certain embodiments, Y.sub.1 is absent or selected from the group consisting of: protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0124] In certain embodiments, Y.sub.1 is absent or selected from the group consisting of: protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0125] In certain embodiments, Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0126] For example, A1 (structural unit

    ##STR00008##

    may be selected from the group consisting of:

    ##STR00009##

    [0127] In certain embodiments, R.sub.1, R.sub.2 and R.sub.3 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.1, R.sub.2 and R.sub.3 comprise a methylene unit, the methylene units of R.sub.1, R.sub.2 and R.sub.3 are each independently unreplaced, or the methylene units of R.sub.1, R.sub.2 and R.sub.3 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0128] In certain embodiments, R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0129] For example, R.sub.1 and R.sub.2 may each independently be selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    [0130] In certain embodiments, R.sub.3 is selected from the group consisting of: hydrogen, optionally substituted OH, optionally substituted SH and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0131] In certain embodiments, when R.sub.3 is optionally substituted methyl, R.sub.3 is substituted with R.sub.31, and R.sub.31 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.31 comprises a methylene unit, the methylene unit of R.sub.31 is each independently unreplaced, or the methylene unit of R.sub.31 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0132] In certain embodiments, R.sub.31 is selected from the group consisting of: hydrogen, halogen, optionally substituted OH and optionally substituted SH.

    [0133] In certain embodiments, when R.sub.31 is optionally substituted OH, R.sub.31 is substituted with R.sub.311, and R.sub.311 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.311 comprises a methylene unit, the methylene unit of R.sub.311 is each independently unreplaced, or the methylene unit of R.sub.311 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0134] In certain embodiments, R.sub.311 is selected from the group consisting of: hydrogen, optionally substituted P(O)H.sub.2, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.2-C.sub.9 heterocyclylene; wherein, when R.sub.311 comprises a methylene unit, the methylene unit of R.sub.311 is each independently unreplaced, or the methylene unit of R.sub.311 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H.sub.2 and optionally substituted NH; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, N+(R).sub.3, PHR, P(R).sub.2, P(O)R.sub.2, OP(O)R.sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0135] In certain embodiments, R.sub.311 is selected from the group consisting of: hydrogen, optionally substituted

    ##STR00010##

    optionally substituted

    ##STR00011##

    optionally substituted

    ##STR00012##

    optionally substituted

    ##STR00013##

    optionally substituted

    ##STR00014##

    optionally substituted

    ##STR00015##

    optionally substituted

    ##STR00016##

    optionally substituted

    ##STR00017##

    optionally substituted

    ##STR00018##

    optionally substituted

    ##STR00019##

    optionally substituted

    ##STR00020##

    optionally substituted

    ##STR00021##

    optionally substituted

    ##STR00022##

    [0136] In certain embodiments, when R.sub.3 is optionally substituted OH, R.sub.3 is substituted with R.sub.32, and R.sub.32 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.32 comprises a methylene unit, the methylene unit of R.sub.32 is each independently unreplaced, or the methylene unit of R.sub.32 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0137] In certain embodiments, R.sub.32 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0138] In certain embodiments, when R.sub.32 is optionally substituted methyl or optionally substituted ethyl, R.sub.32 is substituted with R.sub.321, and R.sub.321 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.321 comprises a methylene unit, the methylene unit of R.sub.321 is each independently unreplaced, or the methylene unit of R.sub.321 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0139] In certain embodiments, R.sub.321 is selected from the group consisting of: hydrogen, halogen, CN and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0140] In certain embodiments, R.sub.321 is selected from the group consisting of: hydrogen, F, Cl, CN and optionally substituted methyl.

    [0141] In certain embodiments, when R.sub.3 is optionally substituted SH, R.sub.3 is substituted with R.sub.33, and R.sub.33 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.33 comprises a methylene unit, the methylene unit of R.sub.33 is each independently unreplaced, or the methylene unit of R.sub.33 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0142] In certain embodiments, R.sub.33 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0143] In certain embodiments, R.sub.33 is optionally substituted methyl.

    [0144] In certain embodiments, when R.sub.33 is optionally substituted methyl, R.sub.33 is substituted with R.sub.331, and R.sub.331 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.331 comprises a methylene unit, the methylene unit of R.sub.331 is each independently unreplaced, or the methylene unit of R.sub.331 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0145] In certain embodiments, R.sub.331 is selected from the group consisting of: hydrogen, chlorine, fluorine and CN.

    [0146] For example, R.sub.3 may be selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted

    ##STR00023##

    optionally substituted

    ##STR00024##

    optionally substituted

    ##STR00025##

    optionally substituted

    ##STR00026##

    optionally substituted

    ##STR00027##

    optionally substituted

    ##STR00028##

    optionally substituted

    ##STR00029##

    optionally substituted

    ##STR00030##

    optionally substituted

    ##STR00031##

    optionally substituted

    ##STR00032##

    optionally substituted

    ##STR00033##

    optionally substituted

    ##STR00034##

    optionally substituted

    ##STR00035##

    optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted OCH.sub.2F, optionally substituted OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    [0147] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described herein, wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl, and R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH,

    ##STR00036##

    OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    [0148] In certain embodiments, R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium and halogen, and R.sub.3 is selected from the group consisting of CH.sub.2OH, SCH.sub.2F,

    ##STR00037##

    [0149] In certain embodiments, B is absent or selected from the group consisting of: optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.8 cycloalkyl, optionally substituted C.sub.3-C.sub.8 heterocyclyl, optionally substituted C.sub.6-C.sub.10 aryl and optionally substituted C.sub.5-C.sub.10 heteroaryl, wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0150] In certain embodiments, B is selected from the group consisting of: optionally substituted

    ##STR00038##

    optionally substituted

    ##STR00039##

    and optionally substituted

    ##STR00040##

    and X.sub.1 is selected from the group consisting of: N and optionally substituted CH.

    [0151] In certain embodiments, when X.sub.1 is optionally substituted CH, X.sub.1 is substituted with R.sub.6, and R.sub.6 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.6 comprises a methylene unit, the methylene unit of R.sub.6 is each independently unreplaced, or the methylene unit of R.sub.6 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0152] In certain embodiments, R.sub.6 is selected from the group consisting of: hydrogen and optionally substituted OH.

    [0153] In certain embodiments, when R.sub.6 is optionally substituted OH, R.sub.6 is substituted with R.sub.61, and R.sub.61 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.61 comprises a methylene unit, the methylene unit of R.sub.61 is each independently unreplaced, or the methylene unit of R.sub.61 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0154] In certain embodiments, R.sub.61 is selected from the group consisting of: H and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0155] In certain embodiments, X.sub.1 is selected from the group consisting of: CH. C(OCH.sub.3) and N.

    [0156] In certain embodiments, B is selected from the group consisting of: [0157] optionally substituted

    ##STR00041##

    optionally substituted

    ##STR00042##

    optionally substituted

    ##STR00043##

    optionally substituted

    ##STR00044##

    optionally substituted

    ##STR00045##

    optionally substituted

    ##STR00046##

    optionally substituted

    ##STR00047##

    optionally substituted

    ##STR00048##

    optionally substituted

    ##STR00049##

    optionally substituted

    ##STR00050##

    optionally substituted

    ##STR00051##

    and optionally substituted

    ##STR00052##

    and X.sub.2 and X.sub.3 are each independently selected from the group consisting of: O, S and optionally substituted NH.

    [0158] In certain embodiments, when X.sub.2 and X.sub.3 are optionally substituted NH, X.sub.2 or X.sub.3 is substituted with R.sub.7, and R.sub.7 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.7 comprises a methylene unit, the methylene unit of R.sub.7 is each independently unreplaced, or the methylene unit of R.sub.7 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0159] In certain embodiments, R.sub.7 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl; wherein, when R.sub.7 comprises a methylene unit, the methylene unit of R.sub.7 is each independently unreplaced, or the methylene unit of R.sub.7 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0160] In certain embodiments, when R.sub.7 is optionally substituted C.sub.1-C.sub.6 alkyl, R.sub.7 is substituted with R.sub.71, and R.sub.71 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.71 comprises a methylene unit, the methylene unit of R.sub.71 is each independently unreplaced, or the methylene unit of R.sub.71 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0161] In certain embodiments, R.sub.71 is selected from the group consisting of: H and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0162] In certain embodiments, X.sub.2 or X.sub.3 is selected from the group consisting of: NH, N(CH.sub.3), O and S.

    [0163] For example, B may be selected from the group consisting of: [0164] optionally substituted

    ##STR00053##

    optionally substituted

    ##STR00054##

    optionally substituted

    ##STR00055##

    optionally substituted

    ##STR00056##

    optionally substituted

    ##STR00057##

    optionally substituted

    ##STR00058##

    optionally substituted

    ##STR00059##

    optionally substituted

    ##STR00060##

    optionally substituted

    ##STR00061##

    optionally substituted

    ##STR00062##

    optionally substituted

    ##STR00063##

    optionally substituted

    ##STR00064##

    optionally substituted

    ##STR00065##

    optionally substituted

    ##STR00066##

    optionally substituted

    ##STR00067##

    optionally substituted

    ##STR00068##

    optionally substituted

    ##STR00069##

    optionally substituted

    ##STR00070##

    optionally substituted

    ##STR00071##

    and optionally substituted

    ##STR00072##

    wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0165] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein B is selected from the group consisting of:

    ##STR00073##

    [0166] In certain embodiments, W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, optionally substituted NHC(O), optionally substituted C(O)NH, optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene; wherein, when W comprises a methylene unit, the methylene unit of W is each independently unreplaced, or the methylene unit of W is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0167] In certain embodiments, when W is optionally substituted C.sub.1-C.sub.6 alkylene, W is substituted with R.sub.8, and R.sub.8 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.8 comprises a methylene unit, the methylene unit of R.sub.8 is each independently unreplaced, or the methylene unit of R.sub.8 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0168] In certain embodiments, R.sub.8 is selected from the group consisting of: hydrogen, O, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted cycloalkyl.

    [0169] In certain embodiments, R.sub.8 is optionally substituted methyl.

    [0170] In certain embodiments, when R.sub.8 is optionally substituted methyl, R.sub.8 is substituted with R.sub.81, and R.sub.81 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

    [0171] In certain embodiments, R.sub.81 is selected from the group consisting of: halogen and optionally substituted cycloalkyl.

    [0172] In certain embodiments, R.sub.81 is selected from the group consisting of: F and optionally substituted cyclopropyl.

    [0173] In certain embodiments, W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted CH.sub.2NHC(O), optionally substituted NHC(O)CH.sub.2, optionally substituted C(O)NH, optionally substituted NH, optionally substituted CHCH, CC, optionally substituted C.sub.1-C.sub.6 alkylene, optionally substituted OCH.sub.2, optionally substituted CH.sub.2O, optionally substituted SCH.sub.2, optionally substituted CH.sub.2S, optionally substituted NHC(O), optionally substituted C(O)CH.sub.2, optionally substituted CH.sub.2NH, optionally substituted NHCH.sub.2, optionally substituted

    ##STR00074##

    optionally substituted

    ##STR00075##

    optionally substituted

    ##STR00076##

    optionally substituted

    ##STR00077##

    and optionally substituted

    ##STR00078##

    [0174] For example, W may be absent or W is selected from the group consisting of:

    ##STR00079##

    [0175] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein W is absent or W is

    ##STR00080##

    [0176] For another example, B may be absent or selected from the group consisting of:

    ##STR00081##

    W may be absent or W is selected from the group consisting of:

    ##STR00082##

    A1 may be selected from the group consisting of:

    ##STR00083##

    [0177] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, which is selected from the group consisting of:

    ##STR00084## [0178] wherein, [0179] X is selected from the group consisting of O, S and NH; [0180] R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl; [0181] N is selected from the group consisting of 1, 2 and 3; [0182] Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; [0183] R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; [0184] R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH,

    ##STR00085##

    OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    [0185] For example, the compound of formula I may be selected from the group consisting of the following structures:

    TABLE-US-00001 No. Structure I-1 [00086]embedded image I-2 [00087]embedded image I-3 [00088]embedded image I-4 [00089]embedded image I-5 [00090]embedded image I-6 [00091]embedded image I-7 [00092]embedded image I-8 [00093]embedded image I-9 [00094]embedded image I-10 [00095]embedded image I-11 [00096]embedded image I-12 [00097]embedded image I-13 [00098]embedded image I-14 [00099]embedded image I-15 [00100]embedded image I-16 [00101]embedded image I-17 [00102]embedded image I-18 [00103]embedded image I-19 [00104]embedded image I-20 [00105]embedded image I-21 [00106]embedded image I-22 [00107]embedded image I-23 [00108]embedded image I-24 [00109]embedded image I-25 [00110]embedded image I-26 [00111]embedded image I-27 [00112]embedded image I-28 [00113]embedded image I-29 [00114]embedded image I-30 [00115]embedded image I-31 [00116]embedded image I-32 [00117]embedded image I-33 [00118]embedded image I-34 [00119]embedded image I-35 [00120]embedded image I-36 [00121]embedded image I-37 [00122]embedded image I-38 [00123]embedded image I-39 [00124]embedded image I-40 [00125]embedded image I-41 [00126]embedded image I-42 [00127]embedded image I-43 [00128]embedded image I-44 [00129]embedded image I-45 [00130]embedded image I-46 [00131]embedded image I-47 [00132]embedded image I-48 [00133]embedded image I-49 [00134]embedded image I-50 [00135]embedded image I-51 [00136]embedded image I-52 [00137]embedded image I-53 [00138]embedded image I-54 [00139]embedded image I-55 [00140]embedded image I-56 [00141]embedded image I-57 [00142]embedded image I-58 [00143]embedded image I-59 [00144]embedded image I-60 [00145]embedded image I-61 [00146]embedded image I-62 [00147]embedded image I-63 [00148]embedded image I-64 [00149]embedded image I-65 [00150]embedded image I-66 [00151]embedded image I-67 [00152]embedded image I-68 [00153]embedded image I-69 [00154]embedded image I-70 [00155]embedded image I-71 [00156]embedded image I-72 [00157]embedded image I-73 [00158]embedded image I-74 [00159]embedded image I-75 [00160]embedded image I-76 [00161]embedded image I-77 [00162]embedded image I-78 [00163]embedded image I-79 [00164]embedded image I-80 [00165]embedded image I-81 [00166]embedded image I-82 [00167]embedded image I-83 [00168]embedded image I-84 [00169]embedded image I-85 [00170]embedded image I-86 [00171]embedded image I-87 [00172]embedded image I-88 [00173]embedded image I-89 [00174]embedded image I-90 [00175]embedded image I-91 [00176]embedded image I-92 [00177]embedded image I-93 [00178]embedded image I-94 [00179]embedded image I-95 [00180]embedded image I-96 [00181]embedded image I-97 [00182]embedded image I-98 [00183]embedded image I-99 [00184]embedded image I-100 [00185]embedded image I-101 [00186]embedded image I-102 [00187]embedded image I-103 [00188]embedded image I-104 [00189]embedded image I-105 [00190]embedded image I-106 [00191]embedded image I-107 [00192]embedded image I-108 [00193]embedded image I-109 [00194]embedded image I-110 [00195]embedded image I-111 [00196]embedded image I-112 [00197]embedded image I-113 [00198]embedded image I-114 [00199]embedded image I-115 [00200]embedded image I-116 [00201]embedded image I-117 [00202]embedded image I-118 [00203]embedded image I-119 [00204]embedded image I-120 [00205]embedded image I-121 [00206]embedded image I-122 [00207]embedded image I-123 [00208]embedded image I-124 [00209]embedded image I-125 [00210]embedded image I-126 [00211]embedded image I-127 [00212]embedded image I-128 [00213]embedded image I-129 [00214]embedded image I-130 [00215]embedded image I-131 [00216]embedded image I-132 [00217]embedded image I-133 [00218]embedded image I-134 [00219]embedded image I-135 [00220]embedded image I-136 [00221]embedded image I-137 [00222]embedded image I-138 [00223]embedded image I-139 [00224]embedded image I-140 [00225]embedded image I-141 [00226]embedded image I-142 [00227]embedded image I-143 [00228]embedded image I-144 [00229]embedded image I-145 [00230]embedded image I-146 [00231]embedded image I-147 [00232]embedded image I-148 [00233]embedded image I-149 [00234]embedded image I-150 [00235]embedded image I-151 [00236]embedded image I-152 [00237]embedded image I-153 [00238]embedded image I-154 [00239]embedded image I-155 [00240]embedded image I-156 [00241]embedded image I-157 [00242]embedded image I-158 [00243]embedded image I-159 [00244]embedded image I-160 [00245]embedded image I-161 [00246]embedded image I-162 [00247]embedded image I-163 [00248]embedded image I-164 [00249]embedded image I-165 [00250]embedded image I-166 [00251]embedded image I-167 [00252]embedded image I-168 [00253]embedded image I-169 [00254]embedded image I-170 [00255]embedded image I-171 [00256]embedded image I-172 [00257]embedded image I-173 [00258]embedded image I-174 [00259]embedded image I-175 [00260]embedded image I-176 [00261]embedded image I-177 [00262]embedded image I-178 [00263]embedded image I-179 [00264]embedded image I-180 [00265]embedded image I-181 [00266]embedded image I-182 [00267]embedded image I-183 [00268]embedded image I-184 [00269]embedded image I-185 [00270]embedded image I-186 [00271]embedded image I-187 [00272]embedded image I-188 [00273]embedded image I-189 [00274]embedded image I-190 [00275]embedded image I-191 [00276]embedded image I-192 [00277]embedded image I-193 [00278]embedded image I-194 [00279]embedded image I-195 [00280]embedded image I-196 [00281]embedded image I-197 [00282]embedded image I-198 [00283]embedded image I-199 [00284]embedded image I-200 [00285]embedded image I-201 [00286]embedded image I-202 [00287]embedded image I-203 [00288]embedded image I-204 [00289]embedded image I-205 [00290]embedded image I-206 [00291]embedded image I-207 [00292]embedded image I-208 [00293]embedded image I-209 [00294]embedded image I-210 [00295]embedded image I-211 [00296]embedded image I-212 [00297]embedded image I-213 [00298]embedded image I-214 [00299]embedded image I-215 [00300]embedded image I-216 [00301]embedded image I-217 [00302]embedded image I-218 [00303]embedded image I-219 [00304]embedded image I-220 [00305]embedded image I-221 [00306]embedded image I-222 [00307]embedded image I-223 [00308]embedded image I-224 [00309]embedded image I-225 [00310]embedded image I-226 [00311]embedded image I-227 [00312]embedded image I-228 [00313]embedded image I-229 [00314]embedded image I-230 [00315]embedded image I-231 [00316]embedded image I-232 [00317]embedded image I-233 [00318]embedded image I-234 [00319]embedded image I-235 [00320]embedded image I-236 [00321]embedded image I-237 [00322]embedded image I-238 [00323]embedded image I-239 [00324]embedded image I-240 [00325]embedded image I-241 [00326]embedded image I-242 [00327]embedded image I-243 [00328]embedded image I-244 [00329]embedded image I-245 [00330]embedded image I-246 [00331]embedded image I-247 [00332]embedded image I-248 [00333]embedded image I-249 [00334]embedded image I-250 [00335]embedded image I-251 [00336]embedded image I-252 [00337]embedded image I-253 [00338]embedded image I-254 [00339]embedded image I-255 [00340]embedded image I-256 [00341]embedded image I-257 [00342]embedded image I-258 [00343]embedded image I-259 [00344]embedded image I-260 [00345]embedded image I-261 [00346]embedded image I-262 [00347]embedded image I-263 [00348]embedded image I-264 [00349]embedded image I-265 [00350]embedded image I-266 [00351]embedded image I-267 [00352]embedded image I-268 [00353]embedded image I-269 [00354]embedded image I-270 [00355]embedded image I-271 [00356]embedded image I-272 [00357]embedded image I-273 [00358]embedded image I-274 [00359]embedded image I-275 [00360]embedded image I-276 [00361]embedded image I-277 [00362]embedded image I-278 [00363]embedded image I-279 [00364]embedded image I-280 [00365]embedded image I-281 [00366]embedded image I-282 [00367]embedded image I-283 [00368]embedded image I-284 [00369]embedded image I-285 [00370]embedded image I-286 [00371]embedded image I-287 [00372]embedded image I-288 [00373]embedded image I-289 [00374]embedded image I-290 [00375]embedded image I-291 [00376]embedded image I-292 [00377]embedded image I-293 [00378]embedded image I-294 [00379]embedded image I-295 [00380]embedded image I-296 [00381]embedded image I-297 [00382]embedded image I-298 [00383]embedded image I-299 [00384]embedded image I-300 [00385]embedded image I-301 [00386]embedded image I-302 [00387]embedded image I-303 [00388]embedded image I-304 [00389]embedded image I-305 [00390]embedded image I-306 [00391]embedded image I-307 [00392]embedded image I-308 [00393]embedded image I-309 [00394]embedded image I-310 [00395]embedded image I-311 [00396]embedded image I-312 [00397]embedded image I-313 [00398]embedded image I-314 [00399]embedded image I-315 [00400]embedded image I-316 [00401]embedded image I-317 [00402]embedded image I-318 [00403]embedded image I-319 [00404]embedded image I-320 [00405]embedded image I-321 [00406]embedded image I-322 [00407]embedded image I-323 [00408]embedded image I-324 [00409]embedded image I-325 [00410]embedded image I-326 [00411]embedded image I-327 [00412]embedded image I-328 [00413]embedded image I-329 [00414]embedded image I-330 [00415]embedded image I-331 [00416]embedded image I-332 [00417]embedded image I-333 [00418]embedded image I-334 [00419]embedded image I-335 [00420]embedded image I-336 [00421]embedded image I-337 [00422]embedded image I-338 [00423]embedded image I-339 [00424]embedded image I-340 [00425]embedded image I-341 [00426]embedded image I-342 [00427]embedded image I-343 [00428]embedded image I-344 [00429]embedded image I-345 [00430]embedded image I-346 [00431]embedded image I-347 [00432]embedded image I-348 [00433]embedded image I-349 [00434]embedded image I-350 [00435]embedded image I-351 [00436]embedded image I-352 [00437]embedded image I-353 [00438]embedded image I-354 [00439]embedded image I-355 [00440]embedded image I-356 [00441]embedded image I-357 [00442]embedded image I-358 [00443]embedded image I-359 [00444]embedded image I-360 [00445]embedded image I-361 [00446]embedded image I-362 [00447]embedded image I-363 [00448]embedded image

    [0186] In another aspect, the present invention provides a compound of a formula below or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

    ##STR00449##

    [0187] In another aspect, the present application provides a compound of formula IIa or IIb:

    ##STR00450##

    or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein, [0188] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0189] B is absent or B is any group; [0190] W is absent or W is any group; [0191] A2 is a substituted benzene ring; [0192] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0193] X is selected from the group consisting of: O, S and NR; [0194] Y.sub.1 is any group, and m is any integer from 0 to 4; [0195] Y.sub.2 is selected from the group consisting of O, S and NR; [0196] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0197] In certain embodiments, the wavy line custom-character in the formula IIa or IIb represents being directly linked to a ligand via the X or Y.sub.2 group, or being linked to the ligand via a Linker fragment.

    [0198] In certain preferred embodiments of the present invention, certain groups in the compounds of formulas (IIa) and (IIb) or pharmaceutically acceptable salts thereof are defined as follows, and groups not mentioned are as described in any one of the embodiments of the present application (in certain embodiments for short).

    [0199] In certain embodiments, CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S and C(O).

    [0200] In certain embodiments, R.sub.4 and R.sub.5 together form optionally substituted cycloalkyl or optionally substituted heterocyclyl. For example, R.sub.4 and R.sub.5 may together form cyclopropyl or cyclobutyl.

    [0201] In certain embodiments, R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0202] In certain embodiments, R.sub.4 and R.sub.5 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.4 and R.sub.5 comprise a methylene unit, the methylene units of R.sub.4 and R.sub.5 are each independently unreplaced, or the methylene units of R.sub.4 and R.sub.5 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0203] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl; and n is selected from the group consisting of 1, 2 and 3.

    [0204] In certain embodiments, Y.sub.1 is selected from the group consisting of: protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0205] In certain embodiments, Y.sub.1 is selected from the group consisting of: protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0206] In certain embodiments, Y.sub.1 is absent or selected from the group consisting of: OR, SR, N(R).sub.2 and optionally substituted C.sub.1-C.sub.6 alkyl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy.

    [0207] In certain embodiments, Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl.

    [0208] In certain embodiments, Y.sub.2 comprises NR; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl, and halogenated C.sub.1-C.sub.6 alkoxy.

    [0209] For example, A2 (structural unit

    ##STR00451##

    may be selected from the group consisting of:

    ##STR00452## ##STR00453## ##STR00454##

    [0210] In certain embodiments, R.sub.1, R.sub.2 and R.sub.3 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.1, R.sub.2 and R.sub.3 comprise a methylene unit, the methylene units of R.sub.1, R.sub.2 and R.sub.3 are each independently unreplaced, or the methylene units of R.sub.1, R.sub.2 and R.sub.3 are each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0211] In certain embodiments, R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0212] In certain embodiments, R.sub.1 and R.sub.2 are each independently selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    [0213] In certain embodiments, R.sub.3 is selected from the group consisting of: hydrogen, optionally substituted OH, optionally substituted SH and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0214] In certain embodiments, when R.sub.3 is optionally substituted methyl, R.sub.3 is substituted with R.sub.31, and R.sub.31 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.31 comprises a methylene unit, the methylene unit of R.sub.31 is each independently unreplaced, or the methylene unit of R.sub.31 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0215] In certain embodiments, R.sub.31 is selected from the group consisting of: hydrogen, halogen, optionally substituted OH and optionally substituted SH.

    [0216] In certain embodiments, when R.sub.31 is optionally substituted OH, R.sub.31 is substituted with R.sub.311, and R.sub.311 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.311 comprises a methylene unit, the methylene unit of R.sub.311 is each independently unreplaced, or the methylene unit of R.sub.311 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0217] In certain embodiments, R.sub.311 is selected from the group consisting of: hydrogen, optionally substituted P(O)H.sub.2, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.2-C.sub.9 heterocyclylene; wherein, when R.sub.311 comprises a methylene unit, the methylene unit of R.sub.311 is each independently unreplaced, or the methylene unit of R.sub.311 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H.sub.2 and optionally substituted NH; wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, N+(R).sub.3, PHR, P(R).sub.2, P(O)R.sub.2, OP(O)R.sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0218] In certain embodiments, R.sub.311 is selected from the group consisting of: hydrogen, optionally substituted

    ##STR00455##

    optionally substituted

    ##STR00456##

    optionally substituted

    ##STR00457##

    optionally substituted

    ##STR00458##

    optionally substituted

    ##STR00459##

    optionally substituted

    ##STR00460##

    optionally substituted

    ##STR00461##

    optionally substituted

    ##STR00462##

    optionally substituted

    ##STR00463##

    optionally substituted

    ##STR00464##

    optionally substituted

    ##STR00465##

    optionally substituted

    ##STR00466##

    optionally substituted

    ##STR00467##

    [0219] In certain embodiments, when R.sub.3 is optionally substituted OH, R.sub.3 is substituted with R.sub.32, and R.sub.32 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.32 comprises a methylene unit, the methylene unit of R.sub.32 is each independently unreplaced, or the methylene unit of R.sub.32 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0220] In certain embodiments, R.sub.32 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0221] In certain embodiments, when R.sub.32 is optionally substituted methyl or optionally substituted ethyl, R.sub.32 is substituted with R.sub.321, and R.sub.321 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.321 comprises a methylene unit, the methylene unit of R.sub.321 is each independently unreplaced, or the methylene unit of R.sub.321 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0222] In certain embodiments, R.sub.321 is selected from the group consisting of: hydrogen, halogen, CN and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0223] In certain embodiments, R.sub.321 is selected from the group consisting of: hydrogen, F, Cl, CN and optionally substituted methyl.

    [0224] In certain embodiments, when R.sub.3 is optionally substituted SH, R.sub.3 is substituted with R.sub.33, and R.sub.33 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.33 comprises a methylene unit, the methylene unit of R.sub.33 is each independently unreplaced, or the methylene unit of R.sub.33 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0225] In certain embodiments, R.sub.33 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0226] In certain embodiments, when R.sub.33 is optionally substituted methyl or optionally substituted ethyl, R.sub.33 is substituted with R.sub.331, and R.sub.331 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.331 comprises a methylene unit, the methylene unit of R.sub.331 is each independently unreplaced, or the methylene unit of R.sub.331 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0227] In certain embodiments, R.sub.331 is selected from the group consisting of: hydrogen, chlorine, fluorine and CN.

    [0228] For example, R.sub.3 may be selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted

    ##STR00468##

    optionally substituted

    ##STR00469##

    optionally substituted

    ##STR00470##

    optionally substituted

    ##STR00471##

    optionally substituted

    ##STR00472##

    optionally substituted

    ##STR00473##

    optionally substituted

    ##STR00474##

    optionally substituted

    ##STR00475##

    optionally substituted

    ##STR00476##

    optionally substituted

    ##STR00477##

    optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted OCH.sub.2F, optionally substituted OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    [0229] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described herein, wherein R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH,

    ##STR00478##

    OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    [0230] In certain embodiments, R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium and halogen, and R.sub.3 is selected from the group consisting of CH.sub.2OH, SCH.sub.2F,

    ##STR00479##

    In certain embodiments, B is absent or selected from the group consisting of: optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.8 cycloalkyl, optionally substituted C.sub.3-C.sub.8 heterocyclyl, optionally substituted C.sub.6-C.sub.10 aryl and optionally substituted C.sub.5-C.sub.10 heteroaryl, wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0231] In certain embodiments, B is selected from the group consisting of: optionally substituted

    ##STR00480##

    optionally substituted

    ##STR00481##

    and optionally substituted

    ##STR00482##

    and X.sub.1 is selected from the group consisting of: N and optionally substituted CH.

    [0232] In certain embodiments, when X.sub.1 is optionally substituted CH, X.sub.1 is substituted with R.sub.6, and R.sub.6 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.6 comprises a methylene unit, the methylene unit of R.sub.6 is each independently unreplaced, or the methylene unit of R.sub.6 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0233] In certain embodiments, R.sub.6 is selected from the group consisting of: hydrogen and optionally substituted OH.

    [0234] In certain embodiments, when R.sub.6 is optionally substituted OH, R.sub.6 is substituted with R.sub.61, and R.sub.61 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.61 comprises a methylene unit, the methylene unit of R.sub.61 is each independently unreplaced, or the methylene unit of R.sub.61 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0235] In certain embodiments, R.sub.61 is selected from the group consisting of: H and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0236] In certain embodiments, X.sub.1 is selected from the group consisting of: CH. C(OCH.sub.3) and N.

    [0237] In certain embodiments, B may be selected from the group consisting of: optionally substituted

    ##STR00483##

    optionally substituted

    ##STR00484##

    optionally substituted

    ##STR00485##

    optionally substituted

    ##STR00486##

    optionally substituted

    ##STR00487##

    optionally substituted

    ##STR00488##

    optionally substituted

    ##STR00489##

    optionally substituted

    ##STR00490##

    optionally substituted

    ##STR00491##

    optionally substituted

    ##STR00492##

    optionally substituted

    ##STR00493##

    optionally substituted

    ##STR00494##

    and optionally substituted

    ##STR00495##

    and X.sub.2 and X.sub.3 are each independently selected from the group consisting of: O, S and optionally substituted NH.

    [0238] In certain embodiments, when X.sub.2 and X.sub.3 are optionally substituted NH, X.sub.2 or X.sub.3 is substituted with R.sub.7, and R.sub.7 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.7 comprises a methylene unit, the methylene unit of R.sub.7 is each independently unreplaced, or the methylene unit of R.sub.7 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0239] In certain embodiments, R.sub.7 is selected from the group consisting of: hydrogen and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0240] In certain embodiments, when R.sub.7 is optionally substituted C.sub.1-C.sub.6 alkyl, R.sub.7 is substituted with R.sub.71, and R.sub.71 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.71 comprises a methylene unit, the methylene unit of R.sub.71 is each independently unreplaced, or the methylene unit of R.sub.71 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0241] In certain embodiments, R.sub.71 is selected from the group consisting of: H and optionally substituted C.sub.1-C.sub.6 alkyl.

    [0242] In certain embodiments, X.sub.2 or X.sub.3 is selected from the group consisting of: NH, N(CH.sub.3), O and S.

    [0243] In certain embodiments, B is selected from the group consisting of: [0244] optionally substituted

    ##STR00496##

    optionally substituted

    ##STR00497##

    optionally substituted

    ##STR00498##

    optionally substituted

    ##STR00499##

    optionally substituted

    ##STR00500##

    optionally substituted

    ##STR00501##

    optionally substituted

    ##STR00502##

    optionally substituted

    ##STR00503##

    optionally substituted

    ##STR00504##

    optionally substituted

    ##STR00505##

    optionally substituted

    ##STR00506##

    optionally substituted

    ##STR00507##

    optionally substituted

    ##STR00508##

    optionally substituted

    ##STR00509##

    optionally substituted

    ##STR00510##

    optionally substituted

    ##STR00511##

    optionally substituted

    ##STR00512##

    optionally substituted

    ##STR00513##

    optionally substituted

    ##STR00514##

    and optionally substituted

    ##STR00515##

    wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0245] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein B is selected from the group consisting of:

    ##STR00516##

    [0246] In certain embodiments, W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, NHC(O), C(O)NH, optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene; wherein, when W comprises a methylene unit, the methylene unit of W is each independently unreplaced, or the methylene unit of W is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0247] In certain embodiments, when W is optionally substituted C.sub.1-C.sub.6 alkylene, W is substituted with R.sub.8, and R.sub.8 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R.sub.8 comprises a methylene unit, the methylene unit of R.sub.8 is each independently unreplaced, or the methylene unit of R.sub.8 is each independently replaced by a group selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted PH, optionally substituted P(O)H, optionally substituted NH, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

    [0248] In certain embodiments, R.sub.8 is selected from the group consisting of: hydrogen, O, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted cycloalkyl.

    [0249] In certain embodiments, R.sub.8 is optionally substituted methyl.

    [0250] In certain embodiments, when R.sub.8 is optionally substituted methyl, R.sub.8 is substituted with R.sub.81, and R.sub.81 is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, O, S, optionally substituted S(O)H, optionally substituted S(O).sub.2H, optionally substituted C(O)H, optionally substituted OH, optionally substituted SH, optionally substituted PH.sub.2, optionally substituted P(O)H.sub.2, optionally substituted NH.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

    [0251] In certain embodiments, R.sub.81 is selected from the group consisting of: halogen and optionally substituted cycloalkyl.

    [0252] In certain embodiments, R.sub.81 is selected from the group consisting of: F and optionally substituted cyclopropyl.

    [0253] In certain embodiments, W is absent or W is selected from the group consisting of: S(O), S(O).sub.2, O, S, C(O), optionally substituted CH.sub.2NHC(O), optionally substituted NHC(O)CH.sub.2, optionally substituted C(O)NH, optionally substituted NH, optionally substituted CHCH, CC, optionally substituted C.sub.1-C.sub.6 alkylene, optionally substituted OCH.sub.2, optionally substituted CH.sub.2O, optionally substituted SCH.sub.2, optionally substituted CH.sub.2S, optionally substituted NHC(O), optionally substituted COCH.sub.2, optionally substituted CH.sub.2NH, optionally substituted NHCH.sub.2, optionally substituted CH(CH.sub.3), optionally substituted

    ##STR00517##

    optionally substituted

    ##STR00518##

    optionally substituted

    ##STR00519##

    optionally substituted

    ##STR00520##

    and optionally substituted

    ##STR00521##

    [0254] For example, W may be absent or W is selected from the group consisting of:

    ##STR00522##

    [0255] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein W is absent or W is

    ##STR00523##

    [0256] For example, B may be absent or selected from the group consisting of:

    ##STR00524##

    W may be absent or W is selected from the group consisting of:

    ##STR00525##

    A2 may be selected from the group consisting of:

    ##STR00526## ##STR00527## ##STR00528##

    [0257] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, which is selected from the group consisting of:

    ##STR00529##

    wherein, [0258] X is selected from the group consisting of O, S and NH; [0259] R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6 alkyl; [0260] N is selected from the group consisting of 1, 2 and 3; [0261] Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; [0262] R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; [0263] R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH,

    ##STR00530##

    OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    [0264] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein the compound is selected from the group consisting of:

    ##STR00531## ##STR00532##

    [0265] In certain embodiments, the compound of formula IIa or IIb or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof further comprises a Trigger fragment (Tr), and the compound comprises the following structure:

    ##STR00533##

    wherein, Tr is selected from the group consisting of the following structures:

    ##STR00534## [0266] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0267] B is absent or B is any group; [0268] W is absent or W is any group; [0269] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0270] X is selected from the group consisting of: O, S and NR; [0271] Y.sub.1 is any group, and m is any integer from 0 to 4; [0272] Y.sub.2 is selected from the group consisting of O, S and NR; [0273] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0274] For example, R.sub.1 and R.sub.2 may each independently be selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    [0275] For example, R.sub.3 may be selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted

    ##STR00535##

    optionally substituted

    ##STR00536##

    optionally substituted

    ##STR00537##

    optionally substituted

    ##STR00538##

    optionally substituted

    ##STR00539##

    optionally substituted

    ##STR00540##

    optionally substituted

    ##STR00541##

    optionally substituted

    ##STR00542##

    optionally substituted

    ##STR00543##

    optionally substituted

    ##STR00544##

    optionally substituted

    ##STR00545##

    optionally substituted

    ##STR00546##

    optionally substituted

    ##STR00547##

    optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted OCH.sub.2F, optionally substituted OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    [0276] For example, B may be selected from the group consisting of: [0277] optionally substituted

    ##STR00548##

    optionally substituted

    ##STR00549##

    optionally substituted

    ##STR00550##

    optionally substituted

    ##STR00551##

    optionally substituted

    ##STR00552##

    optionally substituted

    ##STR00553##

    optionally substituted

    ##STR00554##

    optionally substituted

    ##STR00555##

    optionally substituted

    ##STR00556##

    optionally substituted

    ##STR00557##

    optionally substituted

    ##STR00558##

    optionally substituted

    ##STR00559##

    optionally substituted

    ##STR00560##

    optionally substituted

    ##STR00561##

    optionally substituted

    ##STR00562##

    optionally substituted

    ##STR00563##

    optionally substituted

    ##STR00564##

    optionally substituted

    ##STR00565##

    optionally substituted

    ##STR00566##

    and optionally substituted

    ##STR00567##

    wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0278] For example, W may be absent or W is selected from the group consisting of:

    ##STR00568##

    [0279] In certain embodiments, the Linker fragment comprises an L.sub.1 fragment, an L.sub.2 fragment and/or an L.sub.3 fragment, and the compound has the following structure:

    ##STR00569##

    wherein, [0280] Tr is absent or Tr is any group; [0281] L.sub.3 is selected from a polypeptide fragment; [0282] L.sub.2 is absent or selected from a linker fragment; [0283] L.sub.1 is selected from a coupling unit; [0284] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0285] B is absent or B is any group; [0286] W is absent or W is any group; [0287] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0288] X is selected from the group consisting of: O, S and NR; [0289] Y.sub.1 is any group, and m is any integer from 0 to 4; [0290] Y.sub.2 is selected from the group consisting of O, S and NR; [0291] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0292] In certain preferred embodiments of the present invention, certain groups in the compounds of formulas (IVa) and (IVb) or pharmaceutically acceptable salts thereof are defined as follows, and groups not mentioned are as described in any one of the embodiments of the present application (in certain embodiments for short).

    [0293] In certain embodiments, L.sub.3 is selected from the group consisting of a dipeptide, a tripeptide and a tetrapeptide.

    [0294] In certain embodiments, the dipeptide is selected from the group consisting of: GA, GG, AG, EG, EA, GE, DG, DA, GD, VC, VA, AA and VK.

    [0295] In certain embodiments, the tripeptide is selected from the group consisting of: EAG, EGG, GEG, GEA, DAG, DGG, GDG, GDA, GGA, GAG, GFG, AAG, AAA, VAG, VCG and VKG.

    [0296] In certain embodiments, the tetrapeptide is selected from the group consisting of: GGFG, GGAG, GGGG, GEGG, GEAG, GDGG, GDAG, AAAG and EAGG.

    [0297] In certain embodiments, L.sub.3 is selected from the group consisting of: glycine-glycine-phenylalanine-glycine (GGFG), alanine-alanine-alanine-glycine (AAAG), glycine-glycine-glycine-glycine (GGGG), valine-alanine-glycine (VAG), valine-citrulline-glycine (VCG), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine-alanine (AA), glutamic acid-alanine-glycine-glycine (EAGG), glycine-glutamic acid-alanine-glycine (GEAG), glycine-glutamic acid-glycine-glycine (GEGG), glutamic acid-glycine-glycine (EGG), glutamic acid-alanine-glycine (EAG), valine-lysine-glycine (VKG), glycine-glutamic acid-glycine (GEG), glutamic acid-alanine (EA), glutamic acid-glycine (EG) and glycine-glutamic acid (GE).

    [0298] In certain embodiments, L.sub.2 is absent.

    [0299] In certain embodiments, L.sub.2 comprises or does not comprise a PEG branch chain or a PEG linear chain.

    [0300] In certain embodiments, when L.sub.2 does not comprise a PEG, L.sub.2 is selected from the group consisting of:

    ##STR00570##

    [0301] In certain embodiments, when L.sub.2 comprises the PEG linear chain, L.sub.2 is selected from the group consisting of:

    ##STR00571##

    wherein p is any integer from 1 to 20.

    [0302] In certain embodiments, when L.sub.2 comprises the PEG linear chain, L.sub.2 is selected from the group consisting of:

    ##STR00572##

    [0303] In certain embodiments, when L.sub.2 comprises the PEG branch chain, L.sub.2 is selected from the group consisting of:

    ##STR00573## ##STR00574## ##STR00575##

    wherein q is selected from any integer from 1 to 30.

    [0304] In certain embodiments, when L.sub.2 comprises the PEG branch chain, L.sub.2 is selected from the group consisting of:

    ##STR00576## ##STR00577## ##STR00578## ##STR00579##

    [0305] In certain embodiments, when L.sub.1 is coupled to sulfhydryl of the ligand, L.sub.1 is selected from the group consisting of:

    ##STR00580## ##STR00581##

    wherein, R.sup.L1a, R.sup.L1b and R.sup.L1c are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, OH, SH, NH.sub.2, C(O)H, CO.sub.2H, C(O)C(O)H, C(O)CH.sub.2C(O)H, S(O)H, S(O).sub.2H, C(O)NH.sub.2, SO.sub.2NH.sub.2, OC(O)H, N(H)SO.sub.2H, alkyl, alkenyl, alkynyl, alcyl, heterocyclyl, aryl and heteroaryl.

    [0306] In certain embodiments, R.sup.L1a, R.sup.L1b and R.sup.L1c are each independently selected from the group consisting of: hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted aryl and optionally substituted benzyl.

    [0307] In certain embodiments, when L.sub.1 is coupled to the ligand via amino, L.sub.1 is selected from the group consisting of:

    ##STR00582##

    [0308] In certain embodiments, when L.sub.1 is coupled via click chemistry, L.sub.1 is selected from the group consisting of:

    ##STR00583##

    [0309] For example, the compound of formula IVa or IVb may be selected from the group consisting of the following structures:

    TABLE-US-00002 No. Structure IV- 1 [00584]embedded image IV- 2 [00585]embedded image IV- 6 [00586]embedded image IV- 7 [00587]embedded image IV- 11 [00588]embedded image IV- 12 [00589]embedded image IV- 13 [00590]embedded image IV- 14 [00591]embedded image IV- 15 [00592]embedded image IV- 16 [00593]embedded image IV- 17 [00594]embedded image IV- 21 [00595]embedded image IV- 25 [00596]embedded image IV- 29 [00597]embedded image IV- 30 [00598]embedded image IV- 34 [00599]embedded image IV- 35 [00600]embedded image IV- 39 [00601]embedded image IV- 43 [00602]embedded image IV- 44 [00603]embedded image IV- 45 [00604]embedded image IV- 46 [00605]embedded image IV- 47 [00606]embedded image IV- 48 [00607]embedded image IV- 49 [00608]embedded image IV- 50 [00609]embedded image IV- 51 [00610]embedded image IV- 52 [00611]embedded image IV- 53 [00612]embedded image IV- 54 [00613]embedded image IV- 55 [00614]embedded image IV- 56 [00615]embedded image IV- 57 [00616]embedded image IV- 58 [00617]embedded image IV- 59 [00618]embedded image IV- 60 [00619]embedded image IV- 61 [00620]embedded image IV- 62 [00621]embedded image IV- 63 [00622]embedded image IV- 64 [00623]embedded image IV- 65 [00624]embedded image IV- 66 [00625]embedded image IV- 67 [00626]embedded image IV- 68 [00627]embedded image IV- 69 [00628]embedded image IV- 70 [00629]embedded image IV- 71 [00630]embedded image IV- 81 [00631]embedded image IV- 82 [00632]embedded image IV- 83 [00633]embedded image IV- 84 [00634]embedded image IV- 91 [00635]embedded image IV- 92 [00636]embedded image IV- 93 [00637]embedded image IV- 94 [00638]embedded image IV- 101 [00639]embedded image IV- 102 [00640]embedded image IV- 103 [00641]embedded image IV- 104 [00642]embedded image IV- 111 [00643]embedded image IV- 112 [00644]embedded image IV- 113 [00645]embedded image IV- 114 [00646]embedded image IV- 115 [00647]embedded image IV- 116 [00648]embedded image IV- 117 [00649]embedded image IV- 118 [00650]embedded image IV- 119 [00651]embedded image IV- 120 [00652]embedded image IV- 121 [00653]embedded image IV- 122 [00654]embedded image IV- 123 [00655]embedded image IV- 124 [00656]embedded image IV- 125 [00657]embedded image IV- 126 [00658]embedded image IV- 127 [00659]embedded image IV- 128 [00660]embedded image IV- 129 [00661]embedded image IV- 130 [00662]embedded image IV- 131 [00663]embedded image IV- 141 [00664]embedded image IV- 142 [00665]embedded image IV- 143 [00666]embedded image IV- 144 [00667]embedded image IV- 145 [00668]embedded image IV- 146 [00669]embedded image IV- 147 [00670]embedded image IV- 148 [00671]embedded image IV- 149 [00672]embedded image IV- 150 [00673]embedded image IV- 151 [00674]embedded image IV- 152 [00675]embedded image IV- 153 [00676]embedded image IV- 154 [00677]embedded image IV- 155 [00678]embedded image IV- 156 [00679]embedded image IV- 157 [00680]embedded image IV- 158 [00681]embedded image IV- 159 [00682]embedded image IV- 160 [00683]embedded image

    [0310] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein the compound of formula IVa or IVb is selected from the group consisting of:

    ##STR00684## ##STR00685##

    [0311] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein the compound further comprises a Linker fragment, the compound of formula IIa or IIb is capable of being coupled to the ligand via the Linker fragment, and the compound has the following structure:

    ##STR00686##

    wherein, [0312] Tr is absent or Tr is any group; [0313] L.sub.3 is selected from a polypeptide fragment; [0314] L.sub.2 is absent or selected from a linker fragment; [0315] L.sub.1 is selected from a coupling unit; and L.sub.1 in the formulas IVa-1 and IVb-1 is in a linked form; [0316] R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.8, B, W, CR.sub.4R.sub.8, n, X, Y.sub.1 and Y.sub.2 are each as described in any embodiment of the present invention; [0317] the wavy line custom-character in the general formulas represents being linked to the ligand via the L.sub.1 group.

    [0318] In certain embodiments, provided is the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof of the present invention, wherein the structural unit -Tr-L.sub.3-L.sub.2-L.sub.1- is selected from the group consisting of:

    ##STR00687##

    [0319] In another aspect, the present application provides a conjugate comprising the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described above.

    [0320] In certain embodiments, the conjugate comprises a ligand-drug conjugate. For example, the conjugate may comprise an antibody-drug conjugate.

    [0321] In certain embodiments, the ligand comprises an antibody or an antigen-binding fragment thereof.

    [0322] In certain embodiments, the antibody is selected from the group consisting of: a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, a monoclonal antibody and a polyclonal antibody.

    [0323] In certain embodiments, the antigen-binding fragment is selected from the group consisting of: a Fab, a Fab, a F(ab)2, an Fv, an scFv, a diabody, an Fd, a dAb, a VHH, a maxibody and a complementarity determining region (CDR) fragment.

    [0324] In certain embodiments, the ligand specifically binds to an antigen selected from the group consisting of: AXL, BAFFR, BCMA, BCR-list components, BDCA2, BDCA4, BTLA, BTNL2 BTNL3, BTNL8, BTNL9, C10orf54, CCR1, CCR3, CCR4, CCR5, CCR6, CCR7, CCR9, CCR10, CD11c, CD137, CD138, CD14, CD163, CD168, CD177, CD19, CD20, CD209, CD209L, CD22, CD226, CD248, CD25, CD27, CD274, CD276, CD28, CD30, CD300A, CD33, CD37, CD38, CD4, cluster of differentiation 40 (CD40), CD44, CD45, CD46, CD47, CD48, CD5, CD52, CD55, CD56, CD59, CD62E, CD68, CD69, CD70, CD74, CD79a, CD79b, CD8, CD80, CD86, CD90.2, CD96, CLEC12A, CLEC12B, CLEC7A, CLEC9A, CR1, CR3, CRTAM, CSF1R, CTLA4, CXCR1/2, CXCR4, CXCR5, DDR1, DDR2, DEC-205, DLL4, DR6, FAP, FCamR, FCMR, FcR's, Fire, GITR, HHLA2, HLA class II, HVEM, ICOSLG, IFNAR, type I interferon receptor subunit (IFNAR1), IFNLR1, IL10R1, IL10R2, IL12R, IL13RA1, IL13RA2, IL15R, IL17RA, IL17RB, IL17RC, IL17RE, IL20R1, IL20R2, IL21R, IL22R1, IL22RA, IL23R, IL27R, IL29R, IL2Rg, IL31R, IL36R, IL3RA, IL4R, IL6R, IL5R, IL7R, IL9R, integrins, LAG3, LIFR, MAG/Siglec-4 (sialic acid-binding immunoglobulin-like lectin-4), MMR, MSR1, NCR3LG1, NKG2D, NKp30, NKp46, OX40 (CD134), PDCD1, PROKR1, PVR, PVRIG, PVRL2, PVRL3, RELT, SIGIRR, Siglec-1 (sialic acid-binding immunoglobulin-like lectin-1), Siglec-10, Siglec-5, Siglec-6, Siglec-7, Siglec-8, Siglec-9, SIRPA, SLAMF7, TACI, TCR-list components/assoc, PTCRA, TCRb, CD3z, CD3, TEK, TGFBR1, TGFBR2, TGFBR3, TIGIT, TLR2, TLR4, tumor necrosis factor (TNF), TROY, TSLPR, TYRO, VLDLR, VSIG4, IL2R-y and VTCN1.

    [0325] In certain embodiments, the ligand specifically binds to: TNF, CD40 and/or IFNAR1.

    [0326] In certain embodiments, the ligand is selected from the group consisting of: an anti-TNF antibody or an antigen-binding fragment thereof, an anti-CD40 antibody or an antigen-binding fragment thereof, and an anti-IFNAR1 antibody or an antigen-binding fragment thereof. In certain embodiments, the ligand is selected from the group consisting of: an anti-TNF antibody or an antigen-binding fragment thereof, an anti-CD40 antibody or an antigen-binding fragment thereof, an anti-BDCA2 antibody or an antigen-binding fragment thereof, and an anti-IFNAR1 antibody or an antigen-binding fragment thereof.

    [0327] For example, the ligand may be selected from the group consisting of: an anti-TNF monoclonal antibody, an anti-CD40 monoclonal antibody and an anti-IFNAR1 monoclonal antibody.

    [0328] The present disclosure also provides an immunoconjugate comprising a glucocorticoid receptor agonist linked to an anti-TNF protein. In certain embodiments, the anti-TNF protein is an antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-TNF protein is an antibody or an antigen-binding fragment thereof that binds to TNF (e.g., a soluble TNF and/or a membrane-bound TNF). In certain embodiments, the anti-TNF protein is a soluble TNF receptor protein, e.g., a soluble TNF receptor protein fused to a heavy chain constant domain or a fragment thereof (e.g., Fc). In some embodiments, the anti-TNF protein (e.g., an anti-TNF antibody, an antigen-binding fragment thereof or a soluble TNF receptor) can bind to TNF on the surface of a cell and become internalized. For example, US2014/0294813 (which is incorporated herein by reference in its entirety) discloses an anti-TNF protein that exhibits internalization upon binding to human TNF on the surface of a cell. In certain embodiments, the antibody or the antigen-binding fragment thereof binds to human and/or mouse TNF-. Antibodies and antigen-binding fragments that bind to TNF- are known in the art.

    [0329] Anti-TNF- antibodies and antigen-binding fragments thereof include, for example, adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab, golimumab and anti-mouse TNF mIgG2a. Other anti-TNF- antibodies and antigen-binding fragments are provided in, for example, WO 2013/087912, WO 2014/152247 and WO 2015/073884, each of which is incorporated herein by reference in its entirety.

    [0330] Adalimumab is described in U.S. Pat. No. 6,258,562, which is incorporated herein by reference in its entirety. Infliximab is described in U.S. Pat. No. 5,656,272, which is incorporated herein by reference in its entirety. Certolizumab pegol is discussed in WO 01/94585, which is incorporated herein by reference in its entirety. Afelimomab (also known as MAK195) was discussed in Vincent, Int. J. Clin. Pract. 54: 190-193 (2000), which is incorporated herein by reference in its entirety. Ozoralizumab (also known as ATN-103) is a nanobody. It contains three heavy chain variable regions fused by GlySer linkers. Variable regions 1 and 3 are identical, and ozoralizumab does not contain a heavy chain. Ozoralizumab is discussed in WO 2012/131053, which is incorporated herein by reference in its entirety. Placulumab (also known as CEP-37247) is a domain antibody consisting of a dimer of VL-pCH1-CH2-CH3 or [V-]2-Fc, and is discussed in Gay et al., Mabs 2: 625-638 (2010), which is incorporated herein by reference in its entirety. Golimumab (also known as CNTO 148) is discussed in WO2013/087912, and the sequence is provided in GenBank: D1496971.1 and GenBank DI 496970.1, each of which is incorporated herein by reference in its entirety. Anti-mouse TNF mIgG2a is described in McRae B L et al., J Crohns Colitis 10 (1): 69-76 (2016), each of which is incorporated herein by reference in its entirety.

    [0331] The anti-TNF- antibodies and the antigen-binding fragments thereof also include antibodies and antigen-binding fragments thereof that competitively inhibit the binding of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab to TNF-.

    [0332] The anti-TNF- antibodies and the antigen-binding fragments thereof also include antibodies and antigen-binding fragments that bind to the same TNF- epitope as adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab.

    [0333] In certain embodiments, the anti-TNF- antibody or the antigen-binding fragment thereof competitively inhibits the binding of adalimumab to TNF-. In certain embodiments, the anti-TNF- antibody or the antigen-binding fragment thereof binds to the same TNF- epitope as adalimumab.

    [0334] In certain embodiments, the anti-TNF- antibody or the antigen-binding fragment thereof is adalimumab or an antigen-binding fragment thereof. In certain embodiments, the anti-TNF- antibody or the antigen-binding fragment thereof is adalimumab.

    [0335] In certain embodiments, the anti-TNF- antibody or the antigen-binding fragment thereof comprises a sequence of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab, such as a complementarity determining region (CDR), a variable heavy chain domain (VH), and/or a variable light chain domain (VL).

    [0336] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) of the antibody, wherein the heavy chain variable region comprises an HCDR1, an HCDR2 and an HCDR3 that have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to an HCDR1, an HCDR2 and an HCDR3, respectively, of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0337] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain variable region of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0338] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0339] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain variable region (VL) of the antibody, wherein the light chain variable region comprises an LCDR1, an LCDR2 and an LCDR3 that have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to an LCDR1, an LCDR2 and an LCDR3, respectively, of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0340] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain variable region of the antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0341] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecules: adalimumab, infliximab, afelimomab or golimumab.

    [0342] In certain embodiments, the present application provides an antibody or an antigen-binding fragment thereof, which can specifically bind to TNF- and comprises the Chothia VL CDRs of the VL of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab. In certain aspects, provided herein is an antibody or an antigen-binding fragment thereof, which specifically binds to TNF- and comprises the Chothia VH CDRs of the VH of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab. In certain aspects, provided herein is an antibody or an antigen-binding fragment thereof, which specifically binds to TNF- and comprises the Chothia VL CDRs of the VL of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab and the Chothia VH CDRs of the VH of adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab or golimumab.

    [0343] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) of the antibody, wherein the heavy chain variable region comprises an HCDR1, an HCDR2 and an HCDR3 that have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to an HCDR1, an HCDR2 and an HCDR3, respectively, of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0344] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain variable region of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0345] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0346] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain variable region (VL) of the antibody, wherein the light chain variable region comprises an LCDR1, an LCDR2 and an LCDR3 that have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to an LCDR1, an LCDR2 and an LCDR3, respectively, of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0347] For example, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL) of the antibody, wherein the heavy chain variable region comprises an HCDR1, an HCDR2 and an HCDR3 and the light chain variable region comprises an LCDR1, an LCDR2 and an LCDR3, and these CDRs have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to an HCDR1, an HCDR2, an HCDR3, an LCDR1, an LCDR2 and an LCDR3, respectively, of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0348] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain variable region of the antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0349] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region of the antibody, wherein the heavy chain variable region and the light chain variable region respectively have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0350] In certain embodiments, the anti-TNF antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0351] For example, the anti-TNF antibody or the antigen-binding fragment thereof comprises a heavy chain and a light chain of the antibody, wherein the heavy chain and the light chain respectively have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecules: adalimumab, infliximab, afelimomab, golimumab or anti-mouse TNF mIgG2a 8c11.

    [0352] The sequences of exemplary anti-TNF- antibodies or antigen-binding fragments thereof are provided in Tables 1-2.

    TABLE-US-00003 TABLE1 Aminoacidsequencesforadalimumab LCDR1 RASQGIRNYLA(SEQIDNO:1) LCDR2 AASTLQS(SEQIDNO:2) LCDR3 QRYNRAPYT(SEQIDNO:3) HCDR1 DYAMH(SEQIDNO:4) HCDR2 AITWNSGHIDYADSVEG(SEQIDNO:5) HCDR3 AKVSYLSTASSLDY(SEQIDNO:6) VL DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLL IYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAP YTFGQGTKVEIK(SEQIDNO:7) VH EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDT AVYYCAKVSYLSTASSLDYWGQGTLVTVSS(SEQIDNO:8) Lightchain DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLL IYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAP YTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:9) Heavychain EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDT AVYYCAKVSYLSTASSLDYWGQGTLVTVSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:10)

    TABLE-US-00004 TABLE2 Aminoacidsequencesforanti-mouseTNFmIgG2a8c11 Lightchain QIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWFQQKPGSSPKPWIY ATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSSPLT FGAGTKLELKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDIN VKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNS YTCEATHKTSTSPIVKSFNRNEC(SEQIDNO:31) Heavychain EFQLQQSGPELVKPGASVRISCKASGYSFTDYNMNWVKQSNGKSLE WVGVINPNYGSSTYNQKFKGKATLTVDQSSSTAYMQLNSLTSEDSA VYYCARKWGQLGRGFFDVWGTGTTVTVSSAKTTAPSVYPLAPVCG DTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYT LSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKC PAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWF VNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKV NNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVT DFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKN WVERNSYSCSVVHEGLHNHHTTKSFSRTPGK(SEQIDNO:32)

    [0353] In an embodiment, the antibody or the antigen-binding fragment thereof is an antagonist antibody or an antigen-binding fragment thereof that causes a reduction in CD40 activity or function as compared to CD40 activity or function in the absence of the antibody or the antigen-binding fragment thereof.

    [0354] In particular embodiments, the antibody or the antigen-binding fragment thereof is substantially free of agonist activity, i.e., the antibody or the antigen-binding fragment thereof does not cause an increase in the amount of CD40 activity or function as compared to CD40 activity or function in the absence of the antibody or the antigen-binding fragment thereof. In certain embodiments, the anti-CD40 antibody is a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, or an antigen-binding fragment thereof.

    [0355] In certain embodiments, the anti-CD40 antibody is iscalimab (CFZ533) (Novartis; as described in U.S. Pat. Nos. 8,828,396 and 9,221,913); the anti-CD40 antibody is lucatumumab (Novartis; as described in U.S. Pat. No. 8,277,810); Antibodies 5D12, 3A8 and 3C.sub.6, or humanized versions thereof (Novartis; as described in U.S. Pat. No. 5,874,082); Antibody 15B8 (Novartis; as described in U.S. Pat. No. 7,445,780); Antibody 4D11 (Kyowa Hakko Kirin; as described in U.S. Pat. No. 7,193,064); teneliximab (Bristol Myers Squibb; as described in U.S. Pat. No. 6,051,228); Antibody PG102 (PanGenetics; as described in U.S. Pat. No. 8,669,352); Antibody 2C10 (Primatope; U.S. Patent Application No. 20140093497); anti-CD40 antibodies described in U.S. Pat. Nos. 8,591,900 and 8,778,345 (Boehringer Ingelheim); anti-CD40 antibodies (Amgen) described in U.S. Pat. No. 5,801,227; or APX005 (Boehringer Ingelheim; as described in U.S. Patent Application No. 20120301488.

    [0356] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) of the antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, each of which respectively has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to HCDR1, HCDR2 and HCDR3 of the following molecule: iscalimab (CFZ533).

    [0357] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain variable region of the following molecule: iscalimab (CFZ533).

    [0358] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain of the following molecule: iscalimab (CFZ533).

    [0359] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a light chain variable region (VL) of the antibody, wherein the light chain variable region comprises LCDR1, LCDR2 and LCDR3, each of which respectively has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to LCDR1, LCDR2 and LCDR3 of the following molecule: iscalimab (CFZ533).

    [0360] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a light chain variable region of the antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecule: iscalimab (CFZ533).

    [0361] In certain embodiments, the anti-CD40 antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecule: iscalimab (CFZ533).

    [0362] The sequences of exemplary anti-CD40 antibodies or antigen-binding fragments thereof are provided in Table 3.

    TABLE-US-00005 TABLE3 AminoacidsequencesforCFZ533(Kabat) LCDR1 RSSQSLLYSNGYNYLD(SEQIDNO:11) LCDR2 LGSNRAS(SEQIDNO:12) LCDR3 MQARQTPFT(SEQIDNO:13) HCDR1 SYGMH(SEQIDNO:14) HCDR2 VISYEESNRYHADSVKG(SEQIDNO:15) HCDR3 DGGIAAPGPDY(SEQIDNO:16) VH QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVISYEESNRYHADSVKGRFTISRDNSKITLYLQMNSLRTEDTAV YYCARDGGIAAPGPDYWGQGTLVTVSS(SEQIDNO:17) VL DIVMTQSPLSLTVTPGEPASISCRSSQSLLYSNGYNYLDWYLQKPGQS PQVLISLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ ARQTPFTFGPGTKVDIR(SEQIDNO:18) Lightchain DIVMTQSPLSLTVTPGEPASISCRSSQSLLYSNGYNYLDWYLQKPGQS PQVLISLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQ ARQTPFTFGPGTKVDIRRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:19) Heavychain QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLE WVAVISYEESNRYHADSVKGRFTISRDNSKITLYLQMNSLRTEDTAV YYCARDGGIAAPGPDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:20)

    [0363] In an embodiment, the antibodies of the present invention are specific for (i.e., specifically binds to) IFNAR1. Such antibodies may also be referred to herein as anti-IFNAR1 antibodies of the present invention. In another embodiment, the antibodies of the present invention are specific for human IFNAR1. In another embodiment, the anti-IFNAR1 antibodies of the present invention may cross-react with IFNAR1 of a species other than human or of other proteins structurally related to human IFNAR1 (e.g., human IFNAR1 homologs). In other embodiments, the anti-IFNAR1 antibodies of the present invention may be specific for human IFNAR1 only, and do not exhibit cross-reactivity with species or other types of cross-reactivity.

    [0364] Selection sequences for anti-IFNAR1 antibodies may be found in U.S. Pat. Nos. 5,235,038 and 5,919,453. The entire contents of U.S. Pat. Nos. 10,831,459, 10,182,058, 11,157,494 and 11,521,102 are incorporated herein by reference in their entirety for all purposes.

    [0365] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) of the antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, each of which respectively has at least about 80% sequence identity to HCDR1, HCDR2 and HCDR3 of the following molecule: anifrolumab (MEDI-546).

    [0366] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain variable region of the following molecule: anifrolumab (MEDI-546).

    [0367] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain of the following molecule: anifrolumab (MEDI-546).

    [0368] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a light chain variable region (VL) of the antibody, wherein the light chain variable region comprises LCDR1, LCDR2 and LCDR3, each of which respectively has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to LCDR1, LCDR2 and LCDR3 of the following molecule: anifrolumab (MED-546).

    [0369] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a light chain variable region of the antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecule: anifrolumab (MEDI-546).

    [0370] In certain embodiments, the anti-IFNAR1 antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecule: anifrolumab (MEDI-546).

    [0371] The sequences of exemplary anti-IFNAR1 antibodies or antigen-binding fragments thereof are provided in Table 4.

    TABLE-US-00006 TABLE4 Aminoacidsequencesforanifrolumab(Kabat) LCDR1 RASQSVSSSFFA(SEQIDNO:21) LCDR2 GASSRAT(SEQIDNO:22) LCDR3 QQYDSSAIT(SEQIDNO:23) HCDR1 NYWIA(SEQIDNO:24) HCDR2 IIYPGDSDIRYSPSFQG(SEQIDNO:25) HCDR3 HDIEGFDY(SEQIDNO:26) VH EVQLVQSGAEVKKPGESLKISCKGSGYIFTNYWIAWVRQMPGKGLE SMGIIYPGDSDIRYSPSFQGQVTISADKSITTAYLQWSSLKASDTAMY YCARHDIEGFDYWGRGTLVTVSS(SEQIDNO:27) VL EIVLTQSPGTLSLSPGERATLSCRASQSVSSSFFAWYQQKPGQAPRLLI YGASSRATGIPDRLSGSGSGTDFTLTITRLEPEDFAVYYCQQYDSSAIT FGQGTRLEIK(SEQIDNO:28) Lightchain EIVLTQSPGTLSLSPGERATLSCRASQSVSSSFFAWYQQKPGQAPRLLI YGASSRATGIPDRLSGSGSGTDFTLTITRLEPEDFAVYYCQQYDSSAIT FGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:29) Heavychain EVQLVQSGAEVKKPGESLKISCKGSGYIFTNYWIAWVRQMPGKGLE SMGIIYPGDSDIRYSPSFQGQVTISADKSITTAYLQWSSLKASDTAMY YCARHDIEGFDYWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEFEG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPGK(SEQIDNO:30)

    [0372] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region of the antibody, wherein the heavy chain variable region has at least about 80% sequence identity to a heavy chain variable region of the following molecule: litifilimab (BIIB059). In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a light chain variable region of the antibody, wherein the light chain variable region has at least about 80% sequence identity to a light chain variable region of the following molecule: litifilimab (BIIB059).

    [0373] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80% sequence identity to a heavy chain of the following molecule: litifilimab (BIIB059). In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80% sequence identity to a light chain of the following molecule: litifilimab (BIIB059).

    [0374] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) of the antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain variable region of the following molecule: litifilimab (BIIB059).

    [0375] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a light chain variable region (VL) of the antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain variable region of the following molecule: litifilimab (BIIB059).

    [0376] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a heavy chain of the antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a heavy chain of the following molecule: litifilimab (BIIB059).

    [0377] In certain embodiments, the anti-BDCA2 antibody or the antigen-binding fragment thereof comprises a light chain of the antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to a light chain of the following molecule: litifilimab (BIIB059).

    [0378] The sequences of exemplary anti-BDCA2 antibodies or antigen-binding fragments thereof are provided in Table 5.

    TABLE-US-00007 TABLE5 Aminoacidsequencesforlitifilimab(BIIB059) VL DIQLTQSPSSLSASVGDRVTITCKASQSVDYDGDSYMNWYQQKPGK APKLLIYAASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQA NEDPRTFGQGTKVEIK(SEQIDNO:41) VH DVQLVESGGGLVKPGGSLRLSCAASGFTFSTYTMSWVRQAPGKGLE WVATISPGDSFGYYYPDSVQGRFTISRDNAKNSLYLQMNSLRAEDTA VYYCTRDIYYNYGAWFAYWGQGTLVTVSS(SEQIDNO:36) Lightchain DIQLTQSPSSLSASVGDRVTITCKASQSVDYDGDSYMNWYQQKPGK APKLLIYAASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQA NEDPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQIDNO:42) Heavychain DVQLVESGGGLVKPGGSLRLSCAASGFTFSTYTMSWVRQAPGKGLE WVATISPGDSFGYYYPDSVQGRFTISRDNAKNSLYLQMNSLRAEDTA VYYCTRDIYYNYGAWFAYWGQGTLVTVSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQIDNO:37)

    [0379] The present application also encompasses variants and equivalents that are substantially homologous to the anti-TNF- antibody, the anti-CD40 antibody or the anti-IFNAR1 antibody set forth herein. Such variants and equivalents may contain, for example, conservative substitution mutations, i.e., substitution of one or more amino acids with a similar amino acid. For example, the conservative substitution refers to substitution of an amino acid with another amino acid of the same class, such as substitution of an acidic amino acid with another acidic amino acid, substitution of a basic amino acid with another basic amino acid, or substitution of a neutral amino acid with another neutral amino acid. The purpose of conservative amino acid substitutions is well known in the art. The antibody may be a recombinant polypeptide, a natural polypeptide or a synthetic polypeptide of the antibody. It will be appreciated in the art that some amino acid sequences of the present application may be altered without significantly affecting the structure or function of the protein. Thus, the present application further comprises variants of the polypeptides, which exhibit significant activity or which comprise regions of the antibodies. Such mutants include deletions, insertions, inversions, repeats and type substitutions.

    [0380] In certain embodiments, the ligand is selected from the group consisting of: adalimumab, iscalimab (CFZ533), anifrolumab (MEDI-546), infliximab, afelimomab, golimumab, anti-mouse TNF mIgG2a 8c11, litifilimab (BIIB059), and derivatives and biosimilars thereof.

    [0381] For example, the ligand is selected from the group consisting of: adalimumab, iscalimab (CFZ533) and anifrolumab (MEDI-546).

    [0382] In certain embodiments, the ligand-drug conjugate has the following structure:

    ##STR00688##

    wherein, [0383] Ab represents a ligand capable of binding to a target, including but not limited to, antibodies and antigen-binding fragments thereof, [0384] N.sup.a-I is any number from 1 to 10; [0385] Tr is absent or Tr is any group; [0386] L.sub.3 is selected from a polypeptide fragment; [0387] L.sub.2 is absent or selected from a linker fragment; [0388] L.sub.1 is selected from a coupling unit; [0389] R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently any group; [0390] B is absent or B is any group; [0391] W is absent or W is any group; [0392] CR.sub.4R.sub.5 units are each independently unreplaced or replaced by a group selected from the group consisting of: O, S, NR, S(O), S(O).sub.2, C(O), CC and CC, wherein R.sub.4 and R.sub.5 can together form optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted bridged cyclyl, optionally substituted bridged heterocyclyl, optionally substituted spiro cyclyl and optionally substituted spiro heterocyclyl, and n is any integer from 1 to 20; [0393] X is selected from the group consisting of: O, S and NR; [0394] Y.sub.1 is any group, and m is any integer from 0 to 4; [0395] Y.sub.2 is selected from the group consisting of O, S and NR; [0396] wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0397] The term ligand generally refers to any molecule capable of specifically binding to and/or reactively binding to or complexing a target molecule, such as a receptor, a substrate, an antigenic determinant or other binding site on a target cell or tissue. Examples of ligands include antibodies and fragments thereof (e.g., monoclonal antibodies or fragments thereof), enzymes (e.g., fibrinolytic enzymes), biological response modifiers (e.g., interleukins, interferons, erythropoietins or colony stimulating factors), peptide hormones and antigen-binding fragments thereof, polysaccharides, lipids, oligonucleotides, polynucleotides, synthetic molecules, inorganic molecules, organic molecules, and any combination thereof.

    [0398] The term target or target molecule may include a wide variety of substances and molecules, ranging from simple molecules to complex targets. The target molecules may be proteins, nucleic acids, lipids, carbohydrates, or any other molecules that can be recognized by polypeptide domains. For example, the target molecules may include compounds (i.e., non-biological compounds, e.g., organic molecules, inorganic molecules, or molecules having organic and inorganic atoms, but excluding polynucleotides and proteins), mixtures of compounds, arrays of spatially localized compounds, biological macromolecules, phage peptide display libraries, polysome peptide display libraries, extracts made from biological materials (e.g., bacteria, plants, fungi, or animals (e.g., mammalian) cells or tissues), proteins, toxins, peptide hormones, cells, viruses, and the like. Other target molecules include, for example, whole cells, whole tissues, mixtures of related or unrelated proteins, mixtures of viral or bacterial strains, and the like.

    [0399] The term specifically binding or specific generally refers to a measurable and reproducible interaction, such as binding between a target and an antibody, that may determine the presence of the target in the presence of a heterogeneous population of molecules (including biomolecules). For example, an antibody that specifically binds to a target (which may be an epitope) may be an antibody that binds to the target with greater affinity, avidity, more readily, and/or for greater duration than it binds to other targets. In certain embodiments, the antibody specifically binds to an epitope on the protein that is conserved in proteins of different species. In certain embodiments, specific binding may include, but is not required to be, exclusive binding.

    [0400] The structure of the coupling unit L.sub.1 may be changed before and after coupling to the ligand, i.e., the structure of L.sub.1 may be changed in Linker-Payload structures (e.g., formula IVa or formula IVb) and drug-ligand conjugate structures (e.g., formula Va or formula Vb), and such changes may be easily determined by one of ordinary skill in the art.

    [0401] For example, when L.sub.1 is coupled to the ligand via sulfhydryl, the structural change of L.sub.1 is as follows:

    TABLE-US-00008 Structure before coupling Structure after coupling [00689]embedded image [00690]embedded image [00691]embedded image [00692]embedded image [00693]embedded image [00694]embedded image [00695]embedded image [00696]embedded image [00697]embedded image [00698]embedded image [00699]embedded image [00700]embedded image [00701]embedded image [00702]embedded image [00703]embedded image [00704]embedded image [00705]embedded image [00706]embedded image [00707]embedded image [00708]embedded image [00709]embedded image [00710]embedded image [00711]embedded image [00712]embedded image [00713]embedded image [00714]embedded image
    wherein sulfhydryl may be derived from a ligand, and the R.sup.L1a, R.sup.L1b and R.sup.L1c are independently selected from the group consisting of: hydrogen, optionally substituted alkyl and optionally substituted aryl; for example, the R.sup.L1a, R.sup.L1b and R.sup.L1c may each be independently selected from the group consisting of: hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted aryl and optionally substituted benzyl.

    [0402] For example, when L.sub.1 is coupled to the ligand via amino, the structural change of L.sub.1 is as follows:

    TABLE-US-00009 Structure before coupling Structure after coupling [00715]embedded image [00716]embedded image [00717]embedded image [00718]embedded image
    wherein amino may be derived from a ligand.

    [0403] For example, when L.sub.1 is coupled to a ligand via click chemistry, the structural change of L.sub.1 is as follows:

    TABLE-US-00010 Structure before coupling Structure after coupling [00719]embedded image [00720]embedded image [00721]embedded image [00722]embedded image [00723]embedded image [00724]embedded image [00725]embedded image [00726]embedded image

    [0404] For example, R.sub.1 and R.sub.2 may each independently be selected from the group consisting of: H, F, Cl, Br and optionally substituted methyl.

    [0405] For example, R.sub.3 may be selected from the group consisting of: optionally substituted CH.sub.2Cl, optionally substituted CH.sub.2SH, optionally substituted CH.sub.2OH, optionally substituted

    ##STR00727##

    optionally substituted

    ##STR00728##

    optionally substituted

    ##STR00729##

    optionally substituted

    ##STR00730##

    optionally substituted

    ##STR00731##

    optionally substituted

    ##STR00732##

    optionally substituted

    ##STR00733##

    optionally substituted

    ##STR00734##

    optionally substituted

    ##STR00735##

    optionally substituted

    ##STR00736##

    optionally substituted

    ##STR00737##

    optionally substituted

    ##STR00738##

    optionally substituted

    ##STR00739##

    optionally substituted OH, optionally substituted OCH.sub.3, optionally substituted OCH.sub.2F, optionally substituted OCH.sub.2Cl, optionally substituted OCH.sub.2CN, optionally substituted OCH.sub.2CH.sub.3, optionally substituted sulfhydryl, optionally substituted SCH.sub.2F, optionally substituted SCH.sub.2Cl, optionally substituted SCH.sub.2CF.sub.3 and optionally substituted SCH.sub.2CN.

    [0406] For example, B may be selected from the group consisting of: [0407] optionally substituted

    ##STR00740##

    optionally substituted

    ##STR00741##

    optionally substituted

    ##STR00742##

    optionally substituted

    ##STR00743##

    optionally substituted

    ##STR00744##

    optionally substituted

    ##STR00745##

    optionally substituted

    ##STR00746##

    optionally substituted

    ##STR00747##

    optionally substituted

    ##STR00748##

    optionally substituted

    ##STR00749##

    optionally substituted

    ##STR00750##

    optionally substituted

    ##STR00751##

    optionally substituted

    ##STR00752##

    optionally substituted

    ##STR00753##

    optionally substituted

    ##STR00754##

    optionally substituted

    ##STR00755##

    optionally substituted

    ##STR00756##

    optionally substituted

    ##STR00757##

    optionally substituted

    ##STR00758##

    and optionally substituted

    ##STR00759##

    wherein substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, CN, O, NOH, NOR, NR, OR, C(O)R, C(O)OR, OC(O)R, OC(O)OR, C(O)NHR, C(O)NR.sub.2, OC(O)NHR, OC(O)NR.sub.2, SR, S(O)R, S(O).sub.2R, NHR, N(R).sub.2, NHC(O)R, NRC(O)R, NHC(O)OR, NRC(O)OR, S(O).sub.2NHR, S(O).sub.2N(R).sub.2, NHS(O).sub.2NR.sub.2, NRS(O).sub.2NR.sub.2, NHS(O).sub.2R, NRS(O).sub.2R, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy, wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy.

    [0408] For example, W may be absent or W is selected from the group consisting of:

    ##STR00760##

    [0409] For example, the conjugate may be selected from the group consisting of the following structures:

    TABLE-US-00011 No. Structure V-1 [00761]embedded image V-2 [00762]embedded image V-6 [00763]embedded image V-7 [00764]embedded image V-8 [00765]embedded image V-12 [00766]embedded image V-13 [00767]embedded image V-14 [00768]embedded image V-15 [00769]embedded image V-16 [00770]embedded image V-17 [00771]embedded image V-18 [00772]embedded image V-22 [00773]embedded image V-25 [00774]embedded image V-29 [00775]embedded image V-30 [00776]embedded image V-34 [00777]embedded image V-35 [00778]embedded image V-39 [00779]embedded image V-43 [00780]embedded image V-44 [00781]embedded image V-45 [00782]embedded image V-46 [00783]embedded image V-47 [00784]embedded image V-48 [00785]embedded image V-49 [00786]embedded image V-50 [00787]embedded image V-51 [00788]embedded image V-52 [00789]embedded image V-53 [00790]embedded image V-54 [00791]embedded image V-55 [00792]embedded image V-56 [00793]embedded image V-57 [00794]embedded image V-58 [00795]embedded image V-59 [00796]embedded image V-60 [00797]embedded image V-70 [00798]embedded image V-74 [00799]embedded image V-75 [00800]embedded image V-78 [00801]embedded image V-81 [00802]embedded image V-84 [00803]embedded image V-87 [00804]embedded image V-90 [00805]embedded image V-91 [00806]embedded image V-92 [00807]embedded image V-95 [00808]embedded image V-98 [00809]embedded image V-102 [00810]embedded image V-103 [00811]embedded image V-104 [00812]embedded image V-105 [00813]embedded image V-106 [00814]embedded image V-109 [00815]embedded image V-110 [00816]embedded image V-111 [00817]embedded image V-112 [00818]embedded image V-113 [00819]embedded image V-114 [00820]embedded image V-120 [00821]embedded image V-121 [00822]embedded image V-122 [00823]embedded image V-123 [00824]embedded image V-124 [00825]embedded image V-125 [00826]embedded image V-126 [00827]embedded image V-127 [00828]embedded image V-128 [00829]embedded image V-129 [00830]embedded image V-130 [00831]embedded image V-131 [00832]embedded image V-132 [00833]embedded image V-133 [00834]embedded image V-134 [00835]embedded image V-135 [00836]embedded image V-136 [00837]embedded image V-137 [00838]embedded image V-138 [00839]embedded image V-139 [00840]embedded image V-140 [00841]embedded image V-141 [00842]embedded image V-145 [00843]embedded image V-146 [00844]embedded image V-147 [00845]embedded image V-151 [00846]embedded image V-152 [00847]embedded image V-153 [00848]embedded image V-154 [00849]embedded image V-155 [00850]embedded image V-156 [00851]embedded image V-157 [00852]embedded image V-161 [00853]embedded image V-164 [00854]embedded image V-168 [00855]embedded image V-169 [00856]embedded image V-173 [00857]embedded image V-174 [00858]embedded image V-178 [00859]embedded image V-182 [00860]embedded image V-183 [00861]embedded image V-184 [00862]embedded image V-185 [00863]embedded image V-186 [00864]embedded image V-187 [00865]embedded image V-188 [00866]embedded image V-189 [00867]embedded image V-190 [00868]embedded image V-191 [00869]embedded image V-192 [00870]embedded image V-193 [00871]embedded image V-194 [00872]embedded image V-195 [00873]embedded image V-196 [00874]embedded image V-197 [00875]embedded image V-198 [00876]embedded image V-199 [00877]embedded image V-209 [00878]embedded image V-213 [00879]embedded image V-216 [00880]embedded image V-219 [00881]embedded image V-222 [00882]embedded image V-225 [00883]embedded image V-228 [00884]embedded image V-229 [00885]embedded image V-230 [00886]embedded image V-233 [00887]embedded image V-236 [00888]embedded image V-240 [00889]embedded image V-241 [00890]embedded image V-242 [00891]embedded image V-243 [00892]embedded image V-244 [00893]embedded image V-247 [00894]embedded image V-248 [00895]embedded image V-249 [00896]embedded image V-250 [00897]embedded image V-251 [00898]embedded image V-252 [00899]embedded image V-258 [00900]embedded image V-259 [00901]embedded image V-260 [00902]embedded image V-261 [00903]embedded image V-262 [00904]embedded image V-263 [00905]embedded image V-264 [00906]embedded image V-265 [00907]embedded image V-266 [00908]embedded image V-267 [00909]embedded image V-268 [00910]embedded image V-269 [00911]embedded image V-270 [00912]embedded image V-271 [00913]embedded image V-272 [00914]embedded image V-273 [00915]embedded image V-274 [00916]embedded image V-275 [00917]embedded image V-276 [00918]embedded image V-277 [00919]embedded image V-278 [00920]embedded image V-279 [00921]embedded image V-283 [00922]embedded image V-284 [00923]embedded image V-285 [00924]embedded image V-289 [00925]embedded image V-290 [00926]embedded image V-291 [00927]embedded image V-292 [00928]embedded image V-293 [00929]embedded image V-294 [00930]embedded image V-295 [00931]embedded image V-299 [00932]embedded image V-302 [00933]embedded image V-306 [00934]embedded image V-307 [00935]embedded image V-311 [00936]embedded image V-312 [00937]embedded image V-316 [00938]embedded image V-320 [00939]embedded image V-321 [00940]embedded image V-322 [00941]embedded image V-323 [00942]embedded image V-324 [00943]embedded image V-325 [00944]embedded image V-326 [00945]embedded image V-327 [00946]embedded image V-328 [00947]embedded image V-329 [00948]embedded image V-330 [00949]embedded image V-331 [00950]embedded image V-332 [00951]embedded image V-333 [00952]embedded image V-334 [00953]embedded image V-335 [00954]embedded image V-336 [00955]embedded image V-337 [00956]embedded image V-347 [00957]embedded image V-351 [00958]embedded image V-354 [00959]embedded image V-357 [00960]embedded image V-360 [00961]embedded image V-363 [00962]embedded image V-366 [00963]embedded image V-367 [00964]embedded image V-368 [00965]embedded image V-371 [00966]embedded image V-374 [00967]embedded image V-378 [00968]embedded image V-379 [00969]embedded image V-380 [00970]embedded image V-381 [00971]embedded image V-382 [00972]embedded image V-385 [00973]embedded image V-386 [00974]embedded image V-387 [00975]embedded image V-388 [00976]embedded image V-389 [00977]embedded image V-390 [00978]embedded image V-396 [00979]embedded image V-397 [00980]embedded image V-398 [00981]embedded image V-399 [00982]embedded image V-400 [00983]embedded image V-401 [00984]embedded image V-402 [00985]embedded image V-403 [00986]embedded image V-404 [00987]embedded image V-405 [00988]embedded image V-406 [00989]embedded image V-407 [00990]embedded image V-408 [00991]embedded image V-409 [00992]embedded image V-410 [00993]embedded image V-411 [00994]embedded image V-412 [00995]embedded image V-413 [00996]embedded image V-414 [00997]embedded image V-415 [00998]embedded image
    wherein N.sup.a-I is any number from 1 to 10.

    [0410] In certain embodiments, for the conjugates of the present invention, the ligand-drug conjugates have the following structure:

    ##STR00999##

    wherein, [0411] Ab represents a ligand capable of binding to a target, preferably an antibody or an antigen-binding fragment thereof; [0412] N.sup.a-I is any number from 1 to 10; [0413] X is selected from the group consisting of O, S and NH; [0414] R.sub.4 and R.sub.5 are each independently selected from the group consisting of: H, F, Cl, OH, NH.sub.2 and C.sub.1-C.sub.6alkyl; [0415] N is selected from the group consisting of 1, 2 and 3; [0416] Y.sub.1 is absent or selected from the group consisting of: hydroxy, sulfhydryl, amino and C.sub.1-C.sub.6 alkyl; [0417] R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, fluorine, chlorine and methyl; [0418] R.sub.3 is selected from the group consisting of: CH.sub.2Cl, CH.sub.2SH, CH.sub.2OH,

    ##STR01000##

    OCH.sub.3, OCH.sub.2F, OCH.sub.2Cl, OCH.sub.2CN, OCH.sub.2CH.sub.3, SH, SCH.sub.2F, SCH.sub.2Cl, SCH.sub.2CF.sub.3 and SCH.sub.2CN.

    [0419] In certain embodiments, the conjugates of the present invention are selected from the group consisting of the following structures:

    ##STR01001## ##STR01002##

    wherein N.sup.a-I is any number from 1 to 10, and Ab is selected from an antibody or an antigen-binding fragment thereof.

    [0420] In certain embodiments, the conjugates of the present invention are selected from the group consisting of the following structures:

    ##STR01003## ##STR01004## ##STR01005## ##STR01006##

    wherein N.sup.a-I is any number from 1 to 10.

    [0421] In certain embodiments, the active metabolite of the conjugate comprises the compounds described above.

    [0422] In another aspect, the present application provides a pharmaceutical composition comprising the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described above and/or the conjugate described above, and optionally a pharmaceutically acceptable carrier.

    [0423] In another aspect, the present application provides a method for influencing immune system functions, which comprises administering to a subject the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described above, the conjugate described above and/or the pharmaceutical composition described above.

    [0424] In certain embodiments, the influencing immune system functions comprises influencing immune cell functions.

    [0425] In certain embodiments, the immune cell is selected from the group consisting of: granulocytes and agranulocytes.

    [0426] In certain embodiments, the immune cell is selected from the group consisting of: neutrophils, eosinophils, and basophils.

    [0427] In certain embodiments, the immune cell is selected from the group consisting of: lymphocytes and phagocytes.

    [0428] In certain embodiments, the immune cell is selected from the group consisting of: B cells, T cells, natural killer cells, monocytes, macrophages, mast cells and dendritic cells.

    [0429] In another aspect, the present application provides use of the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described above, the conjugate described above and/or the pharmaceutical composition described above for the manufacture of a medicament for the prevention and/or treatment of a disease and/or condition.

    [0430] In certain embodiments, the disease and/or condition comprises a disease and/or condition associated with glucocorticoid receptor signaling.

    [0431] In certain embodiments, the disease and/or condition is selected from the group consisting of: proliferative diseases and/or conditions, metabolic diseases and/or conditions, inflammatory diseases and/or conditions, and neurodegenerative diseases and/or conditions.

    [0432] In certain embodiments, the disease and/or condition is selected from the group consisting of: systemic autoimmune diseases and/or conditions, blood system-related diseases and/or conditions, neuromuscular system-related diseases and/or conditions, digestive system-related diseases and/or conditions, urological system-related diseases and/or conditions, endocrine gland system-related diseases and/or conditions, cutaneous muscular system-related diseases and/or conditions, and respiratory system-related diseases and/or conditions.

    [0433] In certain embodiments, the disease and/or condition is selected from the group consisting of: rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjogren's syndrome, ankylosing spondylitis, Wegener granulomatosis and systemic sclerosis.

    [0434] In certain embodiments, the disease and/or condition is selected from the group consisting of: autoimmune hemolytic anemia, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia, and vasculitis.

    [0435] In certain embodiments, the disease and/or condition is selected from the group consisting of: multiple sclerosis, myasthenia gravis, and Guillain-Barre syndrome.

    [0436] In certain embodiments, the disease and/or condition is selected from the group consisting of: ulcerative colitis, Crohn's disease, autoimmune liver disease, and atrophic gastritis.

    [0437] In certain embodiments, the disease and/or condition is selected from the group consisting of: IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, Goodpasture's syndrome, and lupus nephritis.

    [0438] In certain embodiments, the disease and/or condition is selected from the group consisting of: type I diabetes, Grave's disease, Hashimoto thyroiditis, primary adrenocortical atrophy and chronic thyroiditis.

    [0439] In certain embodiments, the disease and/or condition is selected from the group consisting of: psoriasis, pemphigus vulgaris, cutaneous lupus erythematosis, dermatomyositis, and polymyalgia rheumatica.

    [0440] In certain embodiments, the disease and/or condition is asthma.

    [0441] In another aspect, the present application provides a method for preventing and/or treating a disease and/or condition, which comprises administering to a subject in need thereof the compound or the tautomer, the mesomer, the racemate, the enantiomer or the diastereoisomer thereof, or the mixture thereof, or the pharmaceutically acceptable salt thereof described above, the conjugate described above and/or the pharmaceutical composition described above.

    [0442] In certain embodiments, the disease and/or condition comprises a disease and/or condition associated with glucocorticoid receptor signaling.

    [0443] In certain embodiments, the disease and/or condition is selected from the group consisting of: proliferative diseases and/or conditions, metabolic diseases and/or conditions, inflammatory diseases and/or conditions, and neurodegenerative diseases and/or conditions.

    [0444] In certain embodiments, the disease and/or condition is selected from the group consisting of: systemic autoimmune diseases and/or conditions, blood system-related diseases and/or conditions, neuromuscular system-related diseases and/or conditions, digestive system-related diseases and/or conditions, urological system-related diseases and/or conditions, endocrine gland system-related diseases and/or conditions, cutaneous muscular system-related diseases and/or conditions, and respiratory system-related diseases and/or conditions.

    [0445] In certain embodiments, the disease and/or condition is selected from the group consisting of: rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjogren's syndrome, ankylosing spondylitis, Wegener granulomatosis and systemic sclerosis.

    [0446] In certain embodiments, the disease and/or condition is selected from the group consisting of: autoimmune hemolytic anemia, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia, and vasculitis.

    [0447] In certain embodiments, the disease and/or condition is selected from the group consisting of: multiple sclerosis, myasthenia gravis, and Guillain-Barre syndrome.

    [0448] In certain embodiments, the disease and/or condition is selected from the group consisting of: ulcerative colitis, Crohn's disease, autoimmune liver disease, and atrophic gastritis.

    [0449] In certain embodiments, the disease and/or condition is selected from the group consisting of: IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, Goodpasture's syndrome, and lupus nephritis.

    [0450] In certain embodiments, the disease and/or condition is selected from the group consisting of: type I diabetes, Grave's disease, Hashimoto thyroiditis, primary adrenocortical atrophy and chronic thyroiditis.

    [0451] In certain embodiments, the disease and/or condition is selected from the group consisting of: psoriasis, pemphigus vulgaris, cutaneous lupus erythematosis, dermatomyositis, and polymyalgia rheumatica.

    [0452] In certain embodiments, the disease and/or condition is asthma. Without being bound by any theory, the following examples are intended only to illustrate the compounds, preparation methods, use, etc., of the present application, and are not intended to limit the scope of the present application.

    EXAMPLES

    [0453] In the present application, [0454] reagents that provide bromination conditions include, but are not limited to, aqueous bromine, N-bromosuccinimide, dibromohydantoin, phosphorus tribromide, liquid bromine/triphenylphosphine, hydrobromic acid, and carbon tetrabromide; [0455] titanium catalysts include, but are not limited to, tetraisopropyl titanate, triisopropoxytitanium chloride, titanium tetrachloride, and triisopropoxytitanium methyl; [0456] palladium catalysts include, but are not limited to, tetrakis(triphenylphosphine)palladium, palladium acetate, palladium chloride, bis(triphenylphosphine)palladium(II) dichloride, tris(dibenzylideneacetone)dipalladium, bis(acetonitrile)palladium(II) dichloride, [1,1-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, [1,1-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex, bis(benzonitrile)palladium chloride, 1,4-butylenebis(diphenylphosphine)-palladium dichloride, allylpalladium chloride dimer, and allyl(cyclopentadienyl)palladium(II); [0457] Borate dimers include, but are not limited to, bis(pinacolato)diboron, bis(neopentyl glycolato)diboron, bis(hexylene glycolato)diboron, bis(catecholato)diboron, bis(diisopropyl-L-tartrate glycolato)diboron, bis[()pinanediol]diborate, bis[(1S,2S,3R,5S)-pinanediolato]diboron, tetramethyidiborane, bis(N,N,N,N-tetramethyl-D-tartramidate)diboron, tetrahydroxydiboron, bis(N,N,N,N-tetramethyl-L-tartramidate)diboron, bis(diisopropyl-D-tartrate glycolato)diboron, bis(diethyl-D-tartrate glycolato)diboron, bis(2,4-dimethyl-2,4-pentanediol)borate, bis(diethyl-L-tartrate glycolato)diboron, and 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-ylboronic acid; [0458] metallic copper salts include, but are not limited to, copper sulfate, copper sulfate pentahydrate, cuprous sulfate, cupric chloride, cuprous chloride, cupric carbonate, cupric phosphate, cupric acetate, and hydrates thereof, cupric oxalate, cupric fluoroborate, and hydrates thereof, cupric methoxide, cupric tartrate, copper formate, cuprous iodide, copper(II) trifluoroacetate, copper(II) trifluoromethanesulfonate, copper carbonate, cupric bromide, cuprous bromide, and cuprous oxide; [0459] the ligand may be selected from any ligand commonly used in the Ullmann reaction, including but not limited to, L-proline, tyrosine, phenylalanine, 1,10-phenanthroline, N,N-dimethylethylenediamine, ethylene glycol, 1,1-binaphthyl-2,2-diol, ethyl 2-cyclohexanonecarboxylate, and salicylaldehyde hydrazone; [0460] the condensing agent may be selected from the group consisting of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N-dicyclohexylcarbodiimide, N,N-diisopropylcarbodiimide, O-benzotriazol-N,N,N,N-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol, O-benzotriazol-N,N,N,N-tetramethyluronium hexafluorophosphate, 2-(7-azobenzotriazol)-N,N,N,N-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, or 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; [0461] reagents that provide basic conditions include organic bases and inorganic bases, wherein the organic bases include, but are not limited to, triethylamine, diethylamine, N-methylmorpholine, pyridine, piperidine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide, potassium tert-butoxide, and the like, and the inorganic bases include, but are not limited to, sodium hydride, potassium carbonate, sodium carbonate, cesium carbonate, sodium hydroxide, lithium hydroxide, sodium phosphate, and potassium phosphate; [0462] reagents that provide acidic conditions include a protic acid and a Lewis acid, wherein the protic acid includes, but is not limited to, hydrochloric acid, sulfuric acid, nitric acid, nitrous acid, sulfurous acid, phosphoric acid, phosphorous acid, formic acid, acetic acid, propionic acid, butyric acid, citric acid, benzoic acid, p-toluenesulfonic acid, p-nitrobenzoic acid, methanesulfonic acid, trifluoromethanesulfonic acid and trifluoroacetic acid, and the Lewis acid includes, but is not limited to, boron trifluoride, zinc chloride, magnesium chloride, aluminum chloride, stannic chloride and ferric chloride; [0463] hydrogenation conditions include, but are not limited to: Pb/C/hydrogen, Pt/C/hydrogen, palladium chloride/hydrogen, Raney nickel/hydrogen, palladium hydroxide carbon/hydrogen, and palladium hydroxide/hydrogen; [0464] reagents that provide oxidation conditions include, but are not limited to, Dess-Martin periodinane, hydrogen peroxide, sodium chlorite, sodium hypochlorite, and potassium perchlorate; [0465] reagents that provide reducing conditions include, but are not limited to, sodium hydride, calcium hydride, lithium hydride, lithium aluminum hydride, sodium borohydride, lithium borohydride, sodium triethylborohydride, sodium triacetoxyborohydride and sodium cyanoborohydride; [0466] reagents that provide oxidation conditions include, but are not limited to, Dess-Martin periodinane, hydrogen peroxide, sodium chlorite, sodium hypochlorite, and potassium perchlorate.

    [0467] In addition, in the present application, [0468] the structure of the compounds is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR is performed using a Quantum-I NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-D), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as solvents, and tetramethylsilane (TMS) as an internal standard, and chemical shifts are given in units of 10_6 (ppm).

    [0469] MS is performed using an Angilent 6230 ESI-TOF mass spectrometer (manufacturer: Agilent, type c: 6230).

    [0470] UPLC is performed using a Waters AcquityUPLCSQD liquid chromatograph-mass spectrometer (Poroshell 120 EC-C18, 2.1 mm50 mm, 1.9 m column).

    [0471] HPLC is performed using an Agilent 1260 high-performance liquid chromatograph (TOSOH G3000 SW SEC column).

    [0472] UV is measured using a Thermo Nanodrop 2000 spectrophotometer.

    [0473] Enzyme-linked immunoassays are performed using an EnVision microplate reader (PerkinElmer).

    [0474] Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate is adopted as a thin layer chromatography (TLC) silica gel plate. The specification adopted by the TLC is 0.15-0.20 mm, and the specification adopted by the thin layer chromatography for the separation and purification of products is 0.4-0.5 mm.

    [0475] Yantai Yellow Sea silica gel of 200-300 mesh is generally utilized as a carrier in column chromatography.

    [0476] Known starting materials of the present application can be synthesized using or according to methods known in the art, or can be purchased from companies such as ABCR GMBH & Co. KG, Acros Organnics, Aldrich Chemical Company, Accela ChemBio Inc., and Darui Chemicals.

    [0477] In the examples, all reactions are carried out under an argon atmosphere or a nitrogen atmosphere unless otherwise stated.

    [0478] The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of argon or nitrogen.

    [0479] The hydrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of hydrogen.

    [0480] In the examples, the solution in the reaction is an aqueous solution unless otherwise stated.

    [0481] In the examples, the reaction temperature is room temperature unless otherwise stated. The room temperature is the optimum reaction temperature, ranging from 20 C. to 30 C.

    [0482] The system of eluents for column chromatography and the system of developing agents for thin layer chromatography used for purifying compounds include: A: dichloromethane and isopropanol system, B: dichloromethane and methanol system, and C: petroleum ether and ethyl acetate system. The volume ratio of solvents is regulated according to different polarities of the compound, and can also be regulated by adding a small amount of triethylamine and acidic or alkaline reagent.

    [0483] Some of the compounds of the present disclosure are characterized by TOF-LC/MS. TOF-LC/MS analysis is performed using an Agilent 6230 time-of-flight mass spectrometer and an Agilent 1290-Infinity ultra-high performance liquid chromatograph.

    [0484] An exemplary preparation route for the present application is as follows:

    I. Preparation of Payload and Linker-Payload

    Preparation Route 1

    ##STR01007## [0485] Step 1: introducing a bromine atom into a benzene ring of a compound of general formula (P1) under optional bromination conditions; [0486] Step 2: reacting a compound of general formula (P2) with a borate dimer under optional palladium reagent catalysis conditions to obtain a compound of general formula (P3); [0487] Step 3: reacting the compound of general formula (P3) with a compound of general formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of general formula (P4); [0488] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0489] Step 5: removing the protecting group PG from the compound of general formula (P5) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0490] In the above steps, [0491] PG may be a common hydroxy protecting group; [0492] Y is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; ring B is optionally substituted aryl or heteroaryl; [0493] B(OR).sub.2 is a boronate monomer in which two R may be linked to form a heterocycle, heterobridged ring or heterospiro ring which may be optionally substituted with C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, carboxy or acyloxy C.sub.1-C.sub.6 alkyl; [0494] X is chlorine, bromine or iodine; [0495] wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl, and halogenated C.sub.1-C.sub.6 alkoxy; [0496] R, R.sub.1, R.sub.2 and R.sub.3 may be defined as R, R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 2

    ##STR01008## [0497] Step 1: reacting a compound of general formula (Y1) with a compound of general formula (P1) under optional metallic copper salt catalysis conditions and optional ligand and under optional basic conditions to obtain a compound of general formula (P2); [0498] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0499] Step 3: reacting the compound of general formula (P3) under optional oxidation conditions to obtain a compound of general formula (P4); [0500] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0501] Step 5: removing the protecting group PG from the compound of general formula (P5) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0502] In the above steps, [0503] PG may be a common hydroxy protecting group; [0504] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; [0505] ring B is optionally substituted aryl or heteroaryl; [0506] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 3

    ##STR01009## [0507] Step 1: subjecting a compound of general formula (P1) to a carbonyl insertion reaction under optional palladium reagent catalysis conditions to obtain a compound of general formula (P2); [0508] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0509] Step 3: reacting the compound of general formula (P3) under optional bromination conditions to obtain a compound of general formula (P4); [0510] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y1) under optional basic conditions to obtain a compound of general formula (P5); [0511] Step 5: reacting the compound of general formula (P5) under optional reducing conditions to obtain a compound of general formula (P6); [0512] Step 6: reacting the compound of general formula (P6) under optional oxidation conditions to obtain a compound of general formula (P7); [0513] Step 7: reacting the compound of general formula (P7) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P8); and [0514] Step 8: removing the protecting group PG from the compound of general formula (P8) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P9).

    [0515] In the above steps, [0516] PG.sub.1 and PG.sub.2 may be common protecting groups for hydroxy or ester groups; [0517] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; R may be optionally substituted C.sub.1-C.sub.6 alkyl; [0518] ring B is optionally substituted aryl or heteroaryl; [0519] R, R.sub.1, R.sub.2 and R.sub.3 may be defined as R, R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 4

    ##STR01010## [0520] Step 1: introducing a bromine atom into methyl of a compound of general formula (P1) under optional bromination conditions to obtain a compound of general formula (P2); [0521] Step 2: reacting the compound of general formula (P2) with a compound of general formula (YT) under optional palladium reagent catalysis conditions to obtain a compound of general formula (P3); [0522] Step 3: reacting the compound of general formula (P3) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P4); and [0523] Step 4: removing the protecting group PG from the compound of general formula (P4) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P5).

    [0524] In the above steps, [0525] PG.sub.1 may be a common hydroxy protecting group; [0526] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; [0527] ring B is optionally substituted aryl or heteroaryl; [0528] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 5

    ##STR01011## [0529] Step 1: reacting a compound of general formula (P1) under optional reducing conditions to obtain a compound of general formula (P2); [0530] Step 2: adding a protecting group PG onto the compound of general formula (P2) under optional acidic or basic conditions to obtain a compound of general formula (P3); [0531] Step 3: reacting the compound of general formula (P3) with a compound of general formula (Y1) under optional basic conditions to obtain a compound of general formula (P4); [0532] Step 4: reacting a compound of general formula (P4) under optional reducing conditions to obtain a compound of general formula (P5); [0533] Step 5: reacting the compound of general formula (P5) under optional oxidation conditions to obtain a compound of general formula (P6); [0534] Step 6: reacting the compound of general formula (P6) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P7); and [0535] Step 7: removing the protecting group PG from the compound of general formula (P7) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P8).

    [0536] In the above steps, [0537] Y is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; [0538] ring B is optionally substituted aryl or heteroaryl; [0539] PG.sub.1 and PG.sub.2 may be common hydroxy or protecting groups of hydroxy; [0540] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 6

    ##STR01012## [0541] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional condensation conditions to obtain a compound of general formula (P2); [0542] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0543] Step 3: reacting the compound of general formula (P3) under optional oxidation conditions to obtain a compound of general formula (P4); [0544] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0545] Step 5: reacting the compound of general formula (P5) under optional reducing conditions to obtain a compound of general formula (P6).

    [0546] In the above steps, [0547] PG.sub.1 and PG.sub.2 may be common protecting groups for hydroxy or ester groups; [0548] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; [0549] ring B is optionally substituted aryl or heteroaryl; [0550] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 7

    ##STR01013## [0551] Step 1: introducing a nitro group into a benzene ring of a compound of general formula (P1) under nitration conditions to obtain a compound of general formula (P2); [0552] Step 2: reacting a compound of general formula (P2) with a borate dimer under optional palladium reagent catalysis conditions to obtain a compound of general formula (P3); [0553] Step 3: reacting the compound of general formula (P3) with a compound of general formula (YT) under optional palladium reagent catalysis conditions to obtain a compound of general formula (P4); [0554] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0555] Step 5: reacting the compound of general formula (P5) under optional hydrogenation conditions to obtain a compound of general formula (P6); and [0556] Step 6: removing the protecting group PG from the compound of general formula (P6) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P7).

    [0557] In the above steps, [0558] PG may be a common hydroxy protecting group; [0559] B(OR).sub.2 is a boronate monomer in which two R may be linked to form a heterocycle, heterobridged ring or heterospiro ring which may be optionally substituted with C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, carboxy or acyloxy C.sub.1-C.sub.6 alkyl; [0560] wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl, and halogenated C.sub.1-C.sub.6 alkoxy; ring B is optionally substituted aryl or heteroaryl; [0561] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Line 8

    ##STR01014## [0562] Step 1: introducing a nitro group into a benzene ring of a compound of general formula (P1) under nitration conditions to obtain a compound of general formula (P2); [0563] Step 2: reacting the compound of general formula (P2) with an ethyl Grignard reagent under optional titanium catalysis conditions to obtain a compound of general formula (P3); [0564] Step 3: reacting a compound of general formula (P2) with a borate dimer under optional palladium reagent catalysis conditions to obtain a compound of general formula (P3); [0565] Step 4: reacting the compound of general formula (P3) with a compound of general formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of general formula (P4); [0566] Step 5: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0567] Step 6: reducing the nitro group of the compound of general formula (P5) under optional hydrogenation conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0568] In the above steps, [0569] ring B is optionally substituted aryl or heteroaryl; [0570] B(OR).sub.2 is a boronate monomer in which two R may be linked to form a heterocycle, heterobridged ring or heterospiro ring which may be optionally substituted with C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, carboxy or acyloxy C.sub.1-C.sub.6 alkyl; [0571] wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl, and halogenated C.sub.1-C.sub.6 alkoxy; [0572] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 9

    ##STR01015## [0573] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional basic conditions to obtain a compound of general formula (P2); [0574] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0575] Step 3: reacting the compound of general formula (P3) under optional oxidation conditions to obtain a compound of general formula (P4); [0576] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0577] Step 5: removing the protecting group PG from the compound of general formula (P5) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0578] In the above steps, [0579] PG may be a common hydroxy protecting group; [0580] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; ring B is optionally substituted aryl or heteroaryl; [0581] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 10

    ##STR01016## [0582] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional palladium catalysis conditions to obtain a compound of general formula (P2); [0583] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0584] Step 3: reacting the compound of general formula (P3) under optional oxidation conditions to obtain a compound of general formula (P4); [0585] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0586] Step 5: removing the protecting group PG from the compound of general formula (P5) under optional acidic or basic conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0587] In the above steps, [0588] X is NR or O, and R is optionally substituted C.sub.1-C.sub.6 alkyl; [0589] PG may be a common hydroxy protecting group; [0590] Y is selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; [0591] ring B is optionally substituted aryl or heteroaryl; [0592] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 11

    ##STR01017## [0593] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional condensation conditions to obtain a compound of general formula (P2); [0594] Step 2: reacting the compound of general formula (P2) under optional reducing conditions to obtain a compound of general formula (P3); [0595] Step 3: reacting the compound of general formula (P3) under optional oxidation conditions to obtain a compound of general formula (P4); [0596] Step 4: reacting the compound of general formula (P4) with a compound of general formula (Y2) under optional acidic or basic conditions to obtain a compound of general formula (P5); and [0597] Step 5: reacting the compound of general formula (P5) under optional reducing conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0598] In the above steps, [0599] PG may be a common hydroxy protecting group; [0600] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, OR, SR, NHR, N(R).sub.2, PHR, P(R).sub.2, P(O)HR, P(O)R.sub.2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogenated C.sub.1-C.sub.6 alkyl and halogenated C.sub.1-C.sub.6 alkoxy; [0601] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application.

    Preparation Route 12

    ##STR01018## [0602] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional condensation conditions to obtain a compound of general formula (P2); [0603] Step 2: removing the protecting group PG.sub.1 from the compound of general formula (P2) under optional acidic or basic conditions to obtain a compound of general formula (P3); [0604] Step 3: reacting the compound of general formula (P3) with a compound of general formula (Y2) under optional condensation conditions to obtain a compound of general formula (P4); [0605] Step 4: removing the protecting group PG.sub.1 from the compound of general formula (P4) under optional acidic or basic conditions to obtain a compound of general formula (P5); [0606] Step 5: reacting the compound of general formula (P5) with a compound of general formula (Y3) under optional condensation conditions to obtain a compound of general formula (P6); [0607] Step 6: removing the protecting group PG1 from the compound of general formula (P6) under optional acidic or basic conditions to obtain a compound of general formula (P7); and [0608] Step 7: reacting the compound of general formula (P7) with a compound of general formula (Y3) under optional condensation conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P8).

    [0609] In the above steps, [0610] ring B is optionally substituted aryl or heteroaryl; [0611] W is absent or W is any group; [0612] X is CH.sub.2, CH(CH.sub.3), CH.sub.2CH.sub.2 or

    ##STR01019## [0613] Y.sub.1 is hydrogen, hydroxy, fluoro, chloro, methyl, hydroxy or methoxy; [0614] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (II), formula (III) or formula (IV) of the present application; [0615] LG.sub.1, LG.sub.2, LG.sub.3 and LG.sub.4 may be common hydroxy of activated ester or carboxy; [0616] PG.sub.1, PG.sub.2 and PG.sub.3 may be common amino protecting groups; [0617] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application. [0618] Tr is absent or Tr is any group; [0619] L.sub.3 is selected from a polypeptide fragment; [0620] L.sub.2 is absent or selected from a linker fragment; [0621] L.sub.1 is selected from a coupling unit.

    Preparation Route 13

    ##STR01020## [0622] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional condensation conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P2).

    [0623] In the above step, [0624] ring B is optionally substituted aryl or heteroaryl; [0625] W is absent or W is any group; [0626] X is CH.sub.2, CH(CH.sub.3), CH.sub.2CH.sub.2 or

    ##STR01021## [0627] Y is O, S or NH; [0628] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application. [0629] LG.sub.1 may be common hydroxy of activated ester or carboxy; [0630] Tr is absent or Tr is any group; [0631] L.sub.3 is selected from a polypeptide fragment; [0632] L.sub.2 is absent or selected from a linker fragment; [0633] L.sub.1 is selected from a coupling unit.

    Preparation Route 14

    ##STR01022## [0634] Step 1: reacting a compound of general formula (P1) with a compound of general formula (Y1) under optional condensation conditions to obtain a compound of general formula (P2); [0635] Step 2: removing the protecting group PG.sub.1 from the compound of general formula (P2) under optional acidic or basic conditions to obtain a compound of general formula (P3); [0636] Step 3: reacting the compound of general formula (P3) with a compound of general formula (Y2) under optional condensation conditions to obtain a compound of general formula (P4); [0637] Step 4: removing the protecting group PG.sub.1 from the compound of general formula (P4) under optional acidic or basic conditions to obtain a compound of general formula (P5); [0638] Step 5: reacting the compound of general formula (P5) with a compound of general formula (Y3) under optional condensation conditions, followed by separation by preparative HPLC, to obtain a compound of general formula (P6).

    [0639] In the above steps, [0640] ring B is optionally substituted aryl or heteroaryl; [0641] W is absent or W is any group; [0642] X is CH.sub.2, CH(CH.sub.3), CH.sub.2CH.sub.2 or

    ##STR01023## [0643] Y is O, S or NH; [0644] LG.sub.1, LG.sub.2 and LG.sub.3 may be common hydroxy of activated ester or carboxy; [0645] PG.sub.1, PG.sub.2 and PG.sub.3 may be common amino protecting groups; [0646] R.sub.1, R.sub.2 and R.sub.3 may be defined as R.sub.1, R.sub.2 and R.sub.3 in any one of formula (I), formula (IIa), formula (IIb), formula (IIIa), formula (IIIb), formula (IVa) or formula (IVb) of the present application. [0647] Tr is absent or Tr is any group; [0648] L.sub.3 is selected from a polypeptide fragment; [0649] L.sub.2 is absent or selected from a linker fragment; [0650] L.sub.1 is selected from a coupling unit.

    II. Preparation of Antibody-Drug Conjugates (ADCs)

    Preparation Route 15

    ##STR01024##

    [0651] The ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0652] wherein, Ab is a ligand containing at least one free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (I-C) or formula (II-C) may be sulfhydryl derived from Ab; [0653] Tr, L.sub.1, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.1, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; L.sub.1x represents the structure after coupling to L.sub.1 coupled to sulfhydryl; [0654] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above.

    Preparation Route 16

    ##STR01025##

    [0655] In Step 1, the ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0656] wherein, Ab is a ligand containing at least one free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (Va) or formula (Vb) may be sulfhydryl derived from Ab; [0657] in Step 2, the compound of any one of formula (Va) or formula (Vb) was incubated in an alkaline buffer at a selected temperature for a selected time to obtain another compound of any one of formula (Va) or formula (Vb) of the present application; [0658] Tr, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; [0659] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above; the selected temperature may be 5 C. to 60 C.; the incubation time may be 0 h to 72 h.

    Preparation Route 17

    ##STR01026##

    [0660] The ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) having a group capable of being coupled to two sulfydryl in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0661] wherein, Ab is a ligand containing at least two free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (Va) or formula (Vb) may be sulfhydryl derived from Ab; [0662] Tr, L.sub.1, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.1, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; L.sub.1y represents the structure after coupling to L.sub.1 coupled to two sulfhydryl; [0663] the compound of formula (Va) or formula (Vb) having a group capable of being coupled to two sulfhydryl may comprise an L.sub.1y group selected from the group consisting of the following:

    ##STR01027##

    wherein each of R.sup.L1a, R.sup.L1b and R.sup.L1c is independently selected from the group consisting of the following: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, OH, SH, NH.sub.2, C(O)H, CO.sub.2H, C(O)C(O)H, C(O)CH.sub.2C(O)H, S(O)H, S(O).sub.2H, C(O)NH.sub.2, SO.sub.2NH.sub.2, OC(O)H, N(H)SO.sub.2H, alkyl, alkenyl, alkynyl, alcyl, heterocyclyl, aryl, and heteroaryl; [0664] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above.

    Preparation Route 18

    ##STR01028##

    [0665] The ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) having a group capable of being coupled to two sulfydryl in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0666] wherein, Ab is a ligand containing at least one free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (Va) or formula (Vb) may be sulfhydryl derived from Ab; [0667] Tr, L.sub.1, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.1, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; L.sub.1x represents the structure after coupling to L.sub.1 coupled to sulfhydryl; [0668] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above.

    Preparation Route 19

    ##STR01029##

    [0669] In Step 1, the ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0670] wherein, Ab is a ligand containing at least one free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (Va) or formula (Vb) may be sulfhydryl derived from Ab; [0671] in Step 2, the compound of any one of formula (Va) or formula (Vb) was incubated in an alkaline buffer at a selected temperature for a selected time to obtain another compound of any one of formula (Va) or formula (Vb) of the present application; [0672] Tr, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; [0673] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above; the selected temperature may be 5 C. to 60 C.; the incubation time may be 0 h to 72 h.

    Preparation Route 20

    ##STR01030##

    [0674] The ligand Ab was reacted with a compound of any one of formula (IVa) or formula (IVb) having a group capable of being coupled to two sulfydryl in an acidic, neutral or alkaline buffer to obtain a compound of formula (Va) or formula (Vb); [0675] wherein, Ab is a ligand containing at least two free sulfhydryl (SH), wherein the free sulfhydryl may be obtained by reducing the ligand by using a reducing agent; the reducing agent includes, but is not limited to, tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, and the like; particularly, disulfide bonds (SS) between ligand chains may be reduced to form free sulfhydryl; wherein the S atom in the compound of formula (Va) or formula (Vb) may be sulfhydryl derived from Ab; [0676] Tr, L.sub.1, L.sub.2 and L.sub.3 may be defined as Tr, L.sub.1, L.sub.2 and L.sub.3 in any one of the compounds of formula (IVa) or formula (IVb) of the present application; Ab and N.sup.a-I may be defined as Ab and N.sup.a-I in any one of the compounds of formula (Va) or formula (Vb) of the present application; L.sub.1y represents the structure after coupling to L.sub.1 coupled to two sulfhydryl; [0677] the compound of formula (Va) or formula (Vb) having a group capable of being coupled to two sulfhydryl may comprise an L.sub.1y group selected from the group consisting of the following:

    ##STR01031##

    wherein each of R.sup.L1a, R.sup.L1b and R.sup.L1c is independently selected from the group consisting of the following: hydrogen, protium, deuterium, tritium, halogen, NO.sub.2, CN, OH, SH, NH.sub.2, C(O)H, CO.sub.2H, C(O)C(O)H, C(O)CH.sub.2C(O)H, S(O)H, S(O).sub.2H, C(O)NH.sub.2, SO.sub.2NH.sub.2, OC(O)H, N(H)SO.sub.2H, alkyl, alkenyl, alkynyl, alcyl, heterocyclyl, aryl, and heteroaryl; [0678] the buffer is selected from the group consisting of the following buffers with a pH of 2 to 12: citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, aqueous ammonia-ammonium chloride buffer, barbiturate sodium-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, and a combination of two or more of the above.

    Example 1

    ##STR01032##

    [0679] Step 1 Compound 1A (500 mg, 3.67 mmol) was dissolved in dichloromethane (10 mL), then DIEA (1.42 g, 10.99 mmol) was added, and TBSCl (830 mg, 5.51 mmol) in dichloromethane (5 mL) was added to the above reaction solution which was then stirred overnight for 17 h. After the reaction was completed as detected by TLC (PE/EA=20/1), the reaction solution was directly dried by rotary evaporation under reduced pressure and subjected to column chromatography (PE:EA=100:0-100:1) to give a milky-white oily liquid 1B (800 mg, yield: 87%).

    [0680] Step 2 Compound 1B (125 mg, 0.50 mmol) and Compound 1C (188 mg, 0.50 mmol) were added to and dissolved in ultra-dry acetonitrile (8 mL), the reaction solution was purged with nitrogen, cooled to 0 C., added with mixture of methanesulfonic acid (48 mg, 0.50 mmol) and MeCN (2 mL) by a syringe, and then stirred overnight for 16 h. After the reaction was completed as detected by TLC (PE/EA=3/1), the reaction solution was added with sodium bicarbonate, filtered, and dried by rotary evaporation to give a crude product which was then purified by preparative chromatography to give Compound 1 (62 mg, yield: 25%) as a white powder and 1S (7 mg, yield: 3%) as a white powder, respectively.

    Compound 1:

    [0681] MS-ESI: m/z 495.3 [M+H].sup.+.

    [0682] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.41 (d, J=8.1 Hz, 2H), 7.34-7.28 (m, 3H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.43 (s, 1H), 4.93 (d, J=4.9 Hz, 1H), 4.79 (brs, 1H), 4.52 (d, J=19.5 Hz, 1H), 4.48 (s, 2H), 4.34-4.25 (m, 1H), 4.19 (d, J=19.5 Hz, 1H), 2.61-2.52 (m, 1H), 2.35-2.27 (m, 1H), 2.19-1.98 (m, 2H), 1.84-1.58 (m, 5H), 1.39 (s, 3H), 1.13-0.95 (m, 2H), 0.87 (s, 3H).

    Compound 1S:

    [0683] MS-ESI: m/z 495.3 [M+H].sup.+.

    [0684] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.35-7.27 (m, 3H), 7.22 (d, J=8.1 Hz, 2H), 6.17 (dd, J=10.1, 1.9 Hz, 1H), 6.09 (s, 1H), 5.95 (s, 1H), 5.30 (d, J=6.8 Hz, 1H), 4.81-4.76 (m, 1H), 4.48 (s, 2H), 4.30 (s, 1H), 4.25 (d, J=19.2 Hz, 1H), 4.01 (d, J=19.2 Hz, 1H), 2.59-2.53 (m, 1H), 2.36-2.28 (m, 1H), 2.12-1.97 (m, 2H), 1.90-1.69 (m, 5H), 1.39 (s, 3H), 1.26-1.13 (m, 1H), 1.06 (dd, J=11.0, 3.4 Hz, 1H), 0.89 (s, 3H).

    ##STR01033##

    [0685] Step 1 Compound 2A (5.0 g, 25.4 mmol) and imidazole (2.59 g, 38.0 mmol) were dissolved in dichloromethane (50 mL), then TBSCl (4.9 g, 32.5 mmol) was added, and the reaction solution was reacted at 25 C. for 2 h. After the reaction was completed as detected by TLC, the reaction solution was added with 100 mL of water and extracted twice with 100 mL of dichloromethane (2), the dichloromethane was combined, dried over anhydrous sodium sulfate, and dried by rotary evaporation, and the residue was directly stirred with silica gel and subjected to column chromatography (PE:EA=20:1) to give a yellow solid 2B (3 g, yield: 38%). MS-ESI: m/z 312.2 [M+H].sup.+.

    [0686] Step 2 Compound 2B (1.0 g, 3.2 mmol) was dissolved in THF (10 mL), the reaction solution was purged with N.sub.2, cooled to 0 C., and then added with BH.sub.3/THF (6.4 mL, 6.4 mmol), and the reaction was completed as detected by LCMS after 17 h. The reaction solution was added to methanol (40 mL) (maintained in an ice bath at 0 C.) to quench BH.sub.3, and then dried by rotary evaporation to give a colorless oil 2C (500 mg, yield: 52%). MS-ESI: m/z 298.1 [M+H].sup.+.

    [0687] Step 3 Compound 2C (500 mg, 1.68 mmol) was dissolved in dichloromethane (8 mL), and the reaction solution was cooled to 0 C., added with Dess-Martin reagent (1.43 g, 3.36 mmol), and reacted for 1.5 h. After the reaction was completed as detected by LCMS, the reaction solution was diluted with water (100 mL) and extracted twice with ethyl acetate (100 mL+50 mL), and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and dried by rotary evaporation to give a white solid 2D (250 mg, yield: 50%). MS-ESI: m/z 296.0 [M+H].sup.+.

    [0688] Step 4 Compound 2D (100 mg, 0.34 mmol), Compound 1C (127 mg, 0.34 mmol) and MgSO.sub.4 (250 mg, 2.08 mmol) were dissolved in acetonitrile (10 mL), the reaction solution was cooled to 0 C. and purged with nitrogen, added with trifluoromethanesulfonic acid (150 mg, 1.0 mmol), and maintained in an ice bath for 2 h after the addition was completed. After the reaction was completed as detected by LCMS, the reaction solution was directly filtered, and the filter cake was rinsed three times with acetonitrile. The mother solution was concentrated by rotary evaporation and subjected to column chromatography (DCM:MeOH=30:1) to give a white solid 2E (100 mg, yield: 55%). MS-ESI: m/z 540.0 [M+H].sup.+.

    [0689] Step 5 Compound 2E (100 mg, 0.18 mmol) and iron powder (100 mg, 1.8 mmol) were added to ethanol (4 mL), NH.sub.4Cl (96 mg, 1.8 mmol) was dissolved in water and added to ethanol, and the reaction solution was purged with N.sub.2, heated to 60 C. and reacted for 2 h. After the reaction was completed as detected by LCMS, the reaction solution was filtered through celite, and the filter cake was rinsed three times with ethanol. The mother solution was concentrated by rotary evaporation and subjected to preparative chromatography and lyophilized to give a white solid powder 2 (40 mg, yield: 42%) and a white solid powder 2S (5 mg, yield: 5%), respectively.

    Compound 2:

    [0690] MS-ESI: m/z 532.2 [M+Na].sup.+.

    [0691] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.32 (d, J=10.1 Hz, 1H), 7.20 (d, J=7.7 Hz, 1H), 6.94 (s, 1H), 6.89 (d, J=7.7 Hz, 1H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.34 (s, 1H), 4.95-4.89 (m, 1H), 4.80 (brs, 1H), 4.49 (d, J=19.4 Hz, 1H), 4.44 (s, 2H), 4.33-4.27 (m, 1H), 4.18 (d, J=19.4 Hz, 1H), 2.62-2.52 (m, 1H), 2.35-2.28 (m, 1H), 2.19-2.07 (m, 1H), 2.07-1.99 (m, 1H), 1.80-1.60 (m, 5H), 1.40 (s, 3H), 1.15-1.02 (m, 1H), 0.99 (dd, J=11.2, 3.6 Hz, 1H), 0.86 (s, 3H).

    Compound 2S:

    [0692] MS-ESI: m/z 532.3 [M+Na].sup.+.

    [0693] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.32 (d, J=10.1 Hz, 1H), 7.15-7.09 (m, 1H), 6.70 (s, 1H), 6.63 (d, J=7.6 Hz, 1H), 6.17 (dd, J=10.0, 1.9 Hz, 1H), 5.99 (s, 1H), 5.94 (s, 1H), 5.25 (d, J=6.8 Hz, 1H), 4.78 (brs, 1H), 4.40 (s, 2H), 4.33-4.22 (m, 2H), 4.02 (d, J=19.2 Hz, 1H), 2.60-2.53 (m, 1H), 2.36-2.28 (m, 1H), 2.11-1.97 (m, 2H), 1.90-1.69 (m, 5H), 1.39 (s, 3H), 1.26-1.13 (m, 1H), 1.09-1.02 (m, 1H), 0.88 (s, 3H).

    ##STR01034##

    [0694] Step 1 Compound 3A (6.58 g, 33.08 mmol, 1.0 eq) and Compound 3B (5.03 g, 33.08 mmol, 1.0 eq) were added into a 250-mL three-necked flask, THF (50 mL), water (5 mL) and K.sub.2CO.sub.3 (13.71 g, 99.24 mmol, 3.0 eq) were added, under nitrogen atmosphere, the reaction solution was added with Pd(dppf)Cl.sub.2 (1.21 g, 1.65 mmol, 0.05 eq), purged three times with nitrogen, heated to 80 C. to reflux, and then stirred for 3-4 h. After the reaction was completed as detected by TLC (PE/EA=5/1), the reaction solution was cooled to room temperature and filtered, and the filtrate was poured into 300 mL of water, and extracted with EA (200 mL). The organic phase was washed with water (100 mL3), washed once with saturated NaCl, dried over anhydrous Na.sub.2SO.sub.4, and concentrated by rotary evaporation, and the residue was stirred with silica gel and subjected to column chromatography (PE/EA=7/1) to give a product which was concentrated by rotary evaporation to give a white solid 3C (2.5 g, yield: 33%).

    [0695] Step 2 Compound 1D (3 g, 7.97 mmol, 1.0 eq), Compound 3C (1.8 g, 7.97 mmol, 1.0 eq) and MgSO.sub.4 (2.88 g, 23.91 mmol, 3.0 eq) were added into a 100-mL three-necked flask, acetonitrile (30 mL) was added, under nitrogen atmosphere, trifluoromethanesulfonic acid (1.2 g, 7.97 mmol, 1.0 eq) was added in an ice-water bath, heated to room temperature and reacted for 2 h after the addition was completed. After the reaction was completed as detected by TLC (PE/EA=3/1), the reaction solution was filtered, the filtrate was concentrated by rotary evaporation, and the residue was stirred with silica gel and subjected to column chromatography (PE/EA=2/1) to give a product which was concentrated by rotary evaporation to give a white solid 3 (3.7 g, yield: 79%).

    [0696] MS-ESI: m/z 585.3 [M+H].sup.+.

    [0697] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.36 (d, J=7.8 Hz, 2H), 7.30 (d, J=10.1 Hz, 1H), 7.24 (d, J=7.7 Hz, 2H), 7.20 (d, J=7.5 Hz, 1H), 7.15 (s, 1H), 7.11 (d, J=7.6 Hz, 1H), 7.07 (d, J=7.6 Hz, 1H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.92 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=4.8 Hz, 1H), 4.78 (brs, 1H), 4.49 (d, J=19.4 Hz, 1H), 4.43 (s, 2H), 4.32-4.25 (m, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.90 (s, 2H), 2.59-2.51 (m, 1H), 2.35-2.26 (m, 1H), 2.18-1.95 (m, 2H), 1.82-1.56 (m, 5H), 1.39 (s, 3H), 1.11-0.96 (m, 4H), 0.85 (s, 3H).

    ##STR01035##

    [0698] Step 1 Compound 4A (500 mg, 2.51 mmol) was dissolved in THF/H.sub.2O (10/2 mL), and then Compound 4B (573 mg, 3.77 mmol) and K.sub.2CO.sub.3 (1.04 g, 7.53 mmol) were added. The reaction solution was purged with nitrogen, then added with Pd(dppf)Cl.sub.2 (367 mg, 0.50 mmol), and heated to 80 C. under nitrogen atmosphere and reacted for 2 h. The reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL3), the organic phases were combined and washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated, and the residue was subjected to column chromatography (PE/EA=1:0 to 3:1) to give a yellow solid 4C (270 mg, yield: 48%).

    [0699] Step 2 Compound 4C (150 mg, 0.66 mmol) was dissolved in MeCN (10 mL), then Compound 1C (250 mg, 0.66 mmol) and MgSO.sub.4 (160 mg, 1.32 mmol) were added, and trifluoromethanesulfonic acid (299 mg, 1.99 mmol) was added at 0 C. under nitrogen atmosphere. The reaction solution was reacted at 0 C. for 1 hour, diluted with water (20 mL), and extracted with dichloromethane (10 mL3), and the organic phases were combined and washed with brine (10 mL), dried, and then concentrated by rotary evaporation to give a crude product which was then subjected to preparative chromatography to give a white powdery solid 4 (120 mg, yield: 31%) and a white powdery solid 4S (13 mg, yield: 3%), respectively.

    Compound 4:

    [0700] MS-ESI: m/z 585.4 [M+H].sup.+.

    [0701] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.35 (d, J=8.0 Hz, 2H), 7.30 (d, J=10.1 Hz, 1H), 7.24-7.16 (m, 4H), 7.14 (d, J=8.1 Hz, 2H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.92 (s, 1H), 5.38 (s, 1H), 4.90 (d, J=4.8 Hz, 1H), 4.48 (d, J=19.4 Hz, 1H), 4.42 (s, 2H), 4.31-4.25 (m, 1H), 4.16 (d, J=19.4 Hz, 1H), 3.89 (s, 2H), 2.59-2.52 (m, 1H), 2.36-2.25 (m, 1H), 2.18-1.96 (m, 2H), 1.82-1.55 (m, 5H), 1.38 (s, 3H), 1.10-0.94 (m, 2H), 0.85 (s, 3H).

    Compound 4S:

    [0702] MS-ESI: m/z 585.3 [M+H].sup.+.

    [0703] .sup.1H NMR (400 MHz, Chloroform-d) 7.31-7.26 (m, 3H), 7.21-7.11 (m, 6H), 6.31 (dd, J=10.1, 1.9 Hz, 1H), 6.08 (s, 2H), 5.43-5.38 (m, 1H), 4.65 (s, 2H), 4.52-4.45 (m, 1H), 4.27 (d, J=19.9 Hz, 1H), 4.05 (d, J=19.9 Hz, 1H), 3.95 (s, 2H), 2.66-2.54 (m, 1H), 2.23-2.07 (m, 3H), 1.95-1.83 (m, 2H), 1.80-1.68 (m, 1H), 1.63 (dd, J=14.1, 2.6 Hz, 1H), 1.46 (s, 3H), 1.31-1.13 (m, 2H), 0.99 (s, 3H).

    ##STR01036##

    [0704] Step 1 Compound 5A (50 g, 216 mmol, 1.0 eq) and imidazole (22 g, 323 mmol, 1.5 eq) were added into a 2-L three-necked flask, the reaction mixture was dissolved in dichloromethane (700 mL), and then added slowly with TBSCl (44 g, 292 mmol, 1.35 eq). After the starting material was consumed completely as detected by TLC (PE/EA=9/1), the reaction solution was cooled to room temperature, filtered, and extracted twice with dichloromethane (100 mL) and H.sub.2O (90 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 5B (65 g, yield: 87%).

    [0705] Step 2 Compound 5B (60 g, 173 mmol, 1.0 eq), bis(pinacolato)diboron (44 g, 173 mmol, 1.0 eq), Pd(dppf)Cl.sub.2 (6.3 g, 8.6 mmol, 0.05 eq) and potassium acetate (51 g, 520 mmol, 3.0 eq) were added into a 2-L three-necked flask, the reaction mixture was dissolved in dioxane (1 L), under N.sub.2 atmosphere, the reaction solution was stirred at 110 C. for 12 h. After the reaction was completed as detected by TLC (PE:EA=9:1), the reaction solution was cooled to room temperature, filtered, and extracted twice with ethyl acetate (1 L) and H.sub.2O (1 L), and the organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 5C (60 g, yield: 88%).

    [0706] Step 3 Compound 5C (30 g, 76 mmol, 1.0 eq), Compound 5D (15.2 g, 76 mmol, 1 eq) and Pd(dppf)Cl.sub.2 (3 g, 4 mmol, 0.05 eq) were added into a 1-L three-necked flask, then dioxane (600 mL) and Cs.sub.2CO.sub.3 (74 g, 228 mmol, 3.0 eq) dissolved in water (60 mL) were added under nitrogen atmosphere, the reaction solution was reacted at 110 C. for 2 h. After the reaction was completed as detected by TLC (PE:EA=9:1), the reaction solution was cooled to room temperature, filtered, and extracted twice with ethyl acetate (1 L) and H.sub.2O (1 L). The organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 5E (20 g, yield: 68%).

    [0707] Step 4 Compound 5E (2.0 g, 5.2 mmol, 1.0 eq), Compound 1C (1.96 g, 5.2 mmol, 1.0 eq) and MgSO.sub.4 (1.88 g, 15.6 mmol, 3.0 eq) were added into a 250-mL three necked flask, the reaction mixture was added with acetonitrile (50 mL), cooled to 20 C. with dry ice and ethanol, added slowly with trifluoromethanesulfonic acid (2.34 g, 15.6 mmol, 3.0 eq), and then stirred at 0 C. for 2 h. After the reaction was completed as detected by TLC (PE:EA=1:1), sodium bicarbonate solution was added to adjust the pH to 7-8, acetonitrile was removed by rotary evaporation, and the reaction solution was extracted and slurried by PE:EA=3:1 to give a crude product of Compound 5F (3.2 g, yield: 98%) which was directly used in the next step. MS-ESI: m/z 630.1 [M+H].sup.+.

    [0708] Step 5 Compound 5F (3.2 g, 5.08 mmol, 1 eq) and Fe (2.84 g, 50.8 mmol, 10 eq) were added into a 250-mL three-necked flask, ethanol (80 mL) was added, NH.sub.4Cl (2.72 g, 50.8 mmol, 10 eq) was dissolved in water (80 mL) and then added into the three-necked flask, and the reaction solution was reacted at 60 C. for 2 h. After the reaction was completed as detected by TLC (DCM:MOH=10:1), ethanol was removed by evaporation under reduced pressure, and aqueous sodium bicarbonate was added to adjust the pH to 8. The reaction solution was extracted twice with 80 mL of dichloromethane and water, and the organic phase was dried over magnesium sulfate, filtered, and concentrated by rotary evaporation to give a yellow solid (2.6 g) as a crude product, 400 mg of which was subjected to reversed-phase column chromatography to give Compound 5 (43.9 mg, purity: 95%).

    [0709] MS-ESI: m/z 600.1 [M+H].sup.+.

    [0710] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.38 (d, J=8.0 Hz, 2H), 7.31 (d, J=10.1 Hz, 1H), 7.24 (d, J=7.9 Hz, 2H), 6.96 (s, 1H), 6.90 (s, 1H), 6.76 (s, 1H), 6.16 (dd, J=10.1, 1.8 Hz, 1H), 5.93 (s, 1H), 5.40 (s, 1H), 4.92 (d, J=4.8 Hz, 1H), 4.79 (brs, 1H), 4.49 (d, J=19.4 Hz, 1H), 4.43 (s, 2H), 4.32-4.26 (m, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.90 (s, 2H), 2.62-2.52 (m, 1H), 2.36-2.26 (m, 1H), 2.18-1.97 (m, 2H), 1.81-1.55 (m, 5H), 1.39 (s, 3H), 1.10-0.94 (m, 2H), 0.86 (s, 3H).

    ##STR01037##

    [0711] Step 1 Compound 6A (10 g, 43.1 mmol) was dissolved in dichloromethane (100 mL), then DIEA (16.7 g, 129.3 mmol) was added, and TBSCl (9.7 g, 64.6 mmol) in dichloromethane (100 mL) was added to the above reaction solution which was then stirred overnight for 3 d. After the reaction was completed as detected by TLC (PE/EA=40/1), the reaction solution was directly dried by rotary evaporation under reduced pressure and subjected to column chromatography with pure petroleum ether to give a light yellow oily liquid 6B (12.0 g, yield: 80%).

    [0712] Step 2 Compound 6B (1 g, 2.89 mmol), bis(pinacolato)diboron (883 mg, 3.48 mmol), KOAc (853 mg, 8.69 mmol) and Pd(dppf)Cl.sub.2 (212 mg, 0.29 mmol) were added to 1,4-dioxane (10 mL), and the reaction was purged with nitrogen, heated (110 C.) and stirred overnight for 17 h. After the reaction was completed as detected by TLC (PE:EA=40:1), the reaction solution was directly dried by rotary evaporation, and the residue was directly stirred with silica gel and subjected to column chromatography (petroleum ether:ethyl acetate=50:1) to give a light yellow oily liquid 6C (183 mg, yield: 16%). MS-ESI: m/z 394.3 [M+H].sup.+.

    [0713] Step 3 Compound 6C (180 mg, 0.46 mmol), Pd(dppf)Cl.sub.2 (33.5 mg, 0.046 mmol), Compound 5D (90.6 mg, 0.46 mg), Cs.sub.2CO.sub.3 (298 mg, 0.92 mmol), 1,4-dioxane (8 mL) and water (2 mL) were added, and the reaction solution was purged with N.sub.2, heated (110 C.) and stirred overnight (20 h). After the reaction was completed as detected by TLC (PE/EA=10/1), the reaction solution was directly dried by rotary evaporation, and the residue was directly stirred with silica gel and subjected to column chromatography (petroleum ether:ethyl acetate=100:1) to give a light yellow oily liquid 6D (95.8 mg, yield: 54%). MS-ESI: m/z 386.1 [M+H].sup.+.

    [0714] Step 4 Compound 6D (500 mg, 1.30 mmol), Compound 1C (488 mg, 1.30 mmol), MeCN (10 mL) and MgSO.sub.4 (930 mg, 7.73 mmol) were added into a three-necked flask, the reaction solution was purged with N.sub.2, cooled to 0 C., added with trifluoromethanesulfonic acid (585 mg, 3.90 mmol) by a constant pressure dropping funnel and stirred (3 h). After the reaction was completed as detected by TLC (DCM/MeOH=10/1), the reaction solution was directly dried by rotary evaporation, and the residue was directly stirred with silica gel and subjected to column chromatography (dichloromethane:MeOH=50:1) to give a white solid 6E (300 mg, yield: 37%); MS-ESI: m/z 630.1 [M+H].sup.+.

    [0715] Step 5 Compound 6E (4 g, 6.35 mmol) and ethanol (100 mL) were added into a single-necked flask, iron powder (2.36 g, 42.1 mmol) and NH.sub.4Cl (3.4 g, 63.6 mmol) were dissolved in water (40 mL) and then added into the above single-necked flask, and the reaction solution was purged with N.sub.2, heated to 60 C. and stirred (2 h). After the reaction was completed as detected by TLC (dichloromethane/CH.sub.3OH=10/1), NaHCO.sub.3 aqueous solution was added to adjust the pH to 7-8, the reaction solution was stirred for 10 min and filtered, the filtrate was concentrated by rotary evaporation, and the residue was added with dichloromethane to dissolve, stirred and subjected to column chromatography (dichloromethane:methanol=30:1) to give a white powdery solid 6 (3.5 g, yield: 92%), 50 mg of which was taken to be subjected to preparative thin layer chromatography and filtered, the filtrate was concentrated by rotary evaporation, and the residue was added with acetonitrile and water (purified water) and lyophilized to give the product 6 (38 mg, purity: 90.08%).

    [0716] MS-ESI: m/z 622.3 [M+Na].sup.+.

    [0717] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.35 (d, J=8.0 Hz, 2H), 7.30 (d, J=10.2 Hz, 1H), 7.22-7.16 (m, 2H), 6.93 (d, J=7.6 Hz, 1H), 6.45-6.40 (m, 1H), 6.40-6.34 (m, 1H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 5.14-5.04 (m, 1H), 4.98-4.83 (m, 3H), 4.78 (d, J=3.4 Hz, 1H), 4.54-4.45 (m, 1H), 4.34-4.26 (m, 3H), 4.22-4.12 (m, 1H), 3.75 (s, 2H), 2.60-2.53 (m, 1H), 2.36-2.26 (m, 1H), 2.17-1.96 (m, 2H), 1.81-1.60 (m, 5H), 1.39 (s, 3H), 1.09-0.97 (m, 2H), 0.85 (s, 3H).

    ##STR01038##

    [0718] Step 1 Compound 7A (65 g, 280 mmol, 1.0 eq) and imidazole (29 g, 420 mmol, 1.5 eq) were added into a 2-L three-necked flask, the reaction mixture was dissolved in dichloromethane (700 mL), and then added slowly with TBSCl (59 g, 392 mmol, 1.35 eq). After the starting material was consumed completely as detected by TLC (PE/EA=9/1), the reaction solution was cooled to room temperature, filtered, and extracted twice with dichloromethane (100 mL) and H.sub.2O (90 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 7B (80 g, yield: 82%).

    [0719] Step 2 Compound 7B (70 g, 203 mmol, 1.0 eq), bis(pinacolato)diboron (51 g, 203 mmol, 1.0 eq), Pd(dppf)Cl.sub.2 (7.4 g, 10.1 mmol, 0.05 eq) and potassium acetate (60 g, 609 mmol, 3.0 eq) were added into a 2-L three-necked flask, the reaction mixture was dissolved in dioxane (1 L), under N.sub.2 atmosphere, the reaction solution was stirred at 110 C. for 12 h. After the reaction was completed as detected by TLC (PE:EA=10:1), the reaction solution was cooled to room temperature, filtered, and extracted twice with ethyl acetate (1 L) and H.sub.2O (1 L), and the organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 7C (60 g, yield: 75%).

    [0720] Step 3 Compound 7C (15 g, 38 mmol, 1.0 eq), Compound 5D (7.6 g, 38 mmol, 1 eq) and Pd(dppf)Cl.sub.2 (1.4 g, 1.9 mmol, 0.05 eq) were added into a 1-L three-necked flask, then dioxane (600 mL) was added, Cs.sub.2CO.sub.3 (37 g, 114 mmol, 3.0 eq) was dissolved in water (50 mL), and under nitrogen atmosphere, the reaction solution was reacted at 110 C. for 2 h. After the reaction was completed as detected by TLC (PE:EA=10:1), the reaction solution was cooled to room temperature, filtered, and extracted twice with ethyl acetate (1 L) and H.sub.2O (1 L). The organic phase was dried over anhydrous sodium sulfate, concentrated by rotary evaporation, stirred with silica gel and purified by normal phase column chromatography (PE:EA=10:1) to give a white solid 7D (10 g, yield: 68%).

    [0721] Step 4 Compound 7D (1 g, 2.6 mmol, 1.0 eq), Compound 1C (0.98 g, 2.6 mmol, 1.0 eq) and MgSO.sub.4 (0.94 g, 7.8 mmol, 3.0 eq) were added into a 250-mL three necked flask, the reaction mixture was added with acetonitrile (50 mL), cooled to 20 C. with dry ice and ethanol, added slowly with trifluoromethanesulfonic acid (1.17 g, 7.8 mmol, 3.0 eq), and then stirred at 0 C. for 2 h. After the reaction was completed as detected by TLC (dichloromethane:MeOH=10:1), sodium bicarbonate solution was added to adjust the pH to 7-8, acetonitrile was removed by rotary evaporation, and the reaction solution was extracted and slurried by PE:EA=3:1 to give Compound 7E (1.4 g, yield: 86%); MS-ESI: m/z 630.1 [M+H].sup.+.

    [0722] Step 5 Compound 7E (200 mg, 0.317 mmol, 1 eq) and iron powder (177 mg, 3.17 mmol, 10 eq) were added into a 25-mL three-necked flask, ethanol (6 mL) was added, NH.sub.4Cl (170 mg, 3.17 mmol, 10 eq) was dissolved in water (6 mL) and then added into the three-necked flask, and the reaction solution was reacted at 60 C. for 2 h. After the reaction was completed as detected by TLC (DCM:MeOH=10:1), ethanol was removed by evaporation under reduced pressure, and aqueous sodium bicarbonate was added to adjust the pH to 8. The reaction solution was extracted twice with 80 mL of dichloromethane and water, and the organic phase was dried over magnesium sulfate, filtered, and concentrated by rotary evaporation to give a yellow solid (200 mg) which was subjected to reversed-phase column chromatography to give Compound 7 (42.6 mg, yield: 22%).

    [0723] MS-ESI: m/z 600.8 [M+H].sup.+.

    [0724] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.36 (d, J=8.0 Hz, 2H), 7.31 (d, J=10.1 Hz, 1H), 7.22 (d, J=7.9 Hz, 2H), 7.20-7.14 (m, 1H), 7.07 (d, J=8.0 Hz, 1H), 6.98 (d, J=8.0 Hz, 1H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=4.7 Hz, 1H), 4.79 (s, 1H), 4.54-4.45 (m, 3H), 4.31-4.26 (m, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.87 (s, 2H), 2.60-2.53 (m, 1H), 2.36-2.26 (m, 1H), 2.18-1.96 (m, 2H), 1.81-1.54 (m, 5H), 1.39 (s, 3H), 1.12-0.94 (m, 2H), 0.86 (s, 3H).

    ##STR01039## ##STR01040##

    [0725] Step 1 Compound 3 (2 g, 3.42 mmol, 1.0 eq) and Compound 8A (2.52 g, 6.84 mmol, 2.0 eq) were added into a reaction flask, and the reaction mixture was dissolved in THF (30 mL), under nitrogen atmosphere, added with TsOH (0.63 g, 3.66 mmol, 1.07 eq) in an ice-water bath, and reacted at 5-10 C. for 30-40 min. After the reaction was completed as detected by TLC (PE/EA=1/2), the reaction solution was added to water, the product was extracted with EA (100 mL), the organic phase was washed with water (50 mL3), dried over by anhydrous sodium sulfate and concentrated, and the residue was stirred with silica gel and subjected to column chromatography (PE/EA=2/1) to give the product which was concentrated by rotary evaporation to give a white solid 8B (2.3 g, yield: 75%).

    [0726] MS-ESI: m/z 893.4 [M+H].sup.+.

    [0727] Step 2 Compound 8B (2 g, 2.24 mmol) was dissolved in DCM (20 mL), and the reaction solution was added dropwise with DEA (6 mL) in an ice-water bath under nitrogen atmosphere, and then reacted at room temperature for 2 h after the addition was completed. After the reaction was completed as detected by TLC (PE/EA=1/2), the reaction solution was concentrated by rotary evaporation, extracted three times with DCM, subjected to reversed-phase column chromatography and lyophilized to give an off-white solid 8C (600 mg, yield: 40%). MS-ESI: m/z 671.3 [M+H].sup.+.

    [0728] Step 3 Compound 8C (300 mg, 0.447 mmol, 1.0 eq) and Compound 8D (230 mg, 0.492 mmol, 1.1 eq) were added into a reaction flask, and the reaction solution was dissolved in DMF (3.5 mL), added with DIEA (173 mg, 1.341 mmol, 3.0 eq) in an ice water-bath under nitrogen atmosphere, then added dropwise with HATU (221 mg, 0.581 mmol, 1.3 eq) in 1 mL DMF, and then stirred at 0-5 C. for 30 min after the addition was completed. After the reaction was completed as detected by LC-MS, the reaction solution was poured into ice water, EA (100 mL) was added, and the product was extracted. The organic phase was washed with water (50 mL3), dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to give a yellow solid 8E (430 mg, yield: 86%). MS-ESI: m/z 1119.4 [M+H].sup.+.

    [0729] Step 4 Under nitrogen atmosphere, Compound 8E (300 mg, 0.268 mmol, 1 eq), Pd(PPh.sub.3).sub.4 (62 mg, 0.0536 mmol, 0.2 eq) and N-methylmorpholine (452 mg, 4.47 mmol, 16.7 eq) were added into a reaction flask, and the reaction mixture was reacted at room temperature for 1 h under nitrogen atmosphere. After the reaction was completed as detected by TLC, the reaction solution was directly purified by reversed-phase column chromatography to give a white solid 8F (210 mg, yield: 72%).

    [0730] MS-ESI: m/z 1079.3 [M+H].sup.+.

    [0731] Step 5 Under nitrogen atmosphere, Compound 8F (200 mg, 0.185 mmol) was dissolved in DCM (2.4 mL), and the reaction solution was added dropwise with DEA (0.8 mL) in an ice-water bath, and heated to room temperature for 2 h after the addition was completed. After the reaction was completed as detected by LCMS, the reaction solution was directly extracted and washed three times with PE (60 mL3), the product precipitated and stuck to the wall of the bottle, and was dissolved with acetonitrile (3 mL), purified by reversed-phase column chromatography to give a white solid 8G (62 mg, yield: 39%). MS-ESI: m/z 857.6 [M+H].sup.+.

    [0732] Step 6 Compound 8G (60 mg, 0.07 mmol, 1.0 eq) was dissolved in THE (3 mL), the reaction solution was dissolved in water (0.6 mL), added with TEA (414 mg, 4.1 mmol, 58 eq) in an ice-water bath, stirred for 1 min, then added with bromoacetyl bromide (113 mg, 0.56 mmol, 8.0 eq), and stirred at 0-5 C. for 5 min. After the reaction was completed as detected by LC-MS, the reaction solution was stored with dry ice and ethanol, subjected to preparative chromatography and lyophilized to give a white solid 8 (11 mg, yield: 16%).

    [0733] MS-ESI: m/z 977.3 [M+H].sup.+.

    [0734] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.62 (t, J=6.6 Hz, 1H), 8.47 (t, J=5.6 Hz, 1H), 8.24 (t, J=5.9 Hz, 1H), 8.18 (d, J=7.7 Hz, 1H), 7.40-7.34 (m, 2H), 7.31 (d, J=10.1 Hz, 1H), 7.27-7.20 (m, 3H), 7.16 (brs, 1H), 7.14-7.07 (m, 2H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=5.1 Hz, 1H), 4.77 (brs, 1H), 4.65-4.55 (m, 2H), 4.49 (d, J=19.4 Hz, 1H), 4.39 (s, 2H), 4.33-4.25 (m, 2H), 4.17 (d, J=19.5 Hz, 1H), 3.94-3.89 (m, 4H), 3.80 (d, J=5.6 Hz, 2H), 3.72 (d, J=5.8 Hz, 2H), 2.57-2.52 (m, 2H), 2.30-2.24 (m, 2H), 2.18-2.06 (m, 1H), 2.06-1.89 (m, 2H), 1.83-1.59 (m, 6H), 1.39 (s, 3H), 1.12-0.97 (m, 2H), 0.85 (s, 3H).

    ##STR01041## ##STR01042##

    [0735] Step 1 Compound 4 (2.4 g, 4.1 mmol, 1.0 eq) and Compound 8A (4.5 g, 12.3 mmol, 3.0 eq) were added into a reaction flask, and the reaction mixture was dissolved in THF (30 mL), under nitrogen atmosphere, added with TsOH (1.1 g, 6.6 mmol, 1.6 eq) in an ice-water bath, and reacted at 5-10 C. for 30-40 min. After the reaction was completed as detected by TLC (PE/EA=1/2), the reaction solution was added to water, the product was extracted with EA (100 mL), the organic phase was washed with water (50 mL3), dried over anhydrous sodium sulfate and concentrated, and the residue was stirred with silica gel and subjected to column chromatography (PE/EA=2/1) to give the product which was concentrated by rotary evaporation to give a white solid 9A (1.5 g, yield: 41%);

    [0736] MS-ESI: m/z 893.4 [M+H].sup.+.

    [0737] Step 2 Compound 9A (1.3 g, 1.46 mmol) was dissolved in DCM (13 mL), and the reaction solution was added dropwise with DEA (2.6 mL) in an ice-water bath under nitrogen atmosphere, and then reacted at room temperature for 2 h after the addition was completed. After the reaction was completed as detected by TLC (PE/EA=1/2), the reaction solution was concentrated by rotary evaporation, extracted three times with DCM, subjected to reversed-phase column chromatography and lyophilized to give an off-white solid 9B (350 mg, yield: 36%). MS-ESI: m/z 670.8 [M+H].sup.+.

    [0738] Step 3 Compound 9B (350 mg, 0.52 mmol, 1.0 eq) and Compound 8D (243 mg, 0.52 mmol, 1.0 eq) were added into a reaction flask, and the reaction solution was dissolved in DMF (3.5 mL), added with DIEA (168 mg, 1.3 mmol, 2.5 eq) in an ice water-bath under nitrogen atmosphere, then added dropwise with HATU (240 mg, 0.63 mmol, 1.21 eq) in 1 mL DMF, and then stirred at 0-5 C. for 30 min after the addition was completed. After the reaction was completed as detected by LC-MS, the reaction solution was poured into ice water, EA (100 mL) was added, and the product was extracted. The organic phase was washed with water (50 mL3), dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to give a foamy yellow solid 9C (600 mg), the yield was not calculated as crude product exceeds theoretic amount; MS-ESI: m/z 1119.4 [M+H].sup.+.

    [0739] Step 4 Under nitrogen atmosphere, Compound 9C (400 mg, 0.35 mmol, 1.0 eq), Pd(PPh.sub.3).sub.4 (82 mg, 0.071 mmol, 0.2 eq) and N-methylmorpholine (353 mg, 3.5 mmol, 10.0 eq) were added into a reaction flask, and the reaction mixture was reacted at room temperature for 1 h under nitrogen atmosphere. After the reaction was completed as detected by LC-MS, the reaction solution was directly purified by reversed-phase column chromatography to give a white solid 9D (150 mg, yield: 39%). MS-ESI: m/z 1079.4 [M+H].sup.+.

    [0740] Step 5 Under nitrogen atmosphere, Compound 9D (150 mg, 0.14 mmol) was dissolved in DCM (1.5 mL), and the reaction solution was added dropwise with DEA (0.3 mL) in an ice-water bath, and heated to room temperature for 2 h after the addition was completed. After the reaction was completed as detected by LCMS, the reaction solution was directly concentrated by rotary evaporation, extracted three times with DCM, and purified by reversed-phase column chromatography to give a white solid 9E (60 mg, yield: 50%). MS-ESI: m/z 857.4 [M+H].sup.+.

    [0741] Step 6 Compound 9E (50 mg, 0.058 mmol, 1.0 eq) was dissolved in THE (3 mL), the reaction solution was added with 3 drops of water for dissolving, added with TEA (117 mg, 1.16 mmol, 20.0 eq) in an ice-water bath, stirred for 1 min, then added with bromoacetyl bromide (93 mg, 0.46 mmol, 8.0 eq), and stirred at 0-5 C. for 5 min. After the reaction was completed as detected by LC-MS, the reaction solution was stored with dry ice, subjected to preparative chromatography and lyophilized to give a white solid 9 (11 mg, yield: 19%);

    [0742] MS-ESI: m/z 977.3 [M+H].sup.+.

    [0743] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.62 (t, J=6.6 Hz, 1H), 8.49 (t, J=5.6 Hz, 1H), 8.26 (t, J=5.8 Hz, 1H), 8.19 (d, J=7.5 Hz, 1H), 7.36 (d, J=7.9 Hz, 2H), 7.30 (d, J=10.1 Hz, 1H), 7.26-7.14 (m, 6H), 6.16 (dd, J=10.1, 1.9 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=4.9 Hz, 1H), 4.79 (brs, 1H), 4.61-4.54 (m, 2H), 4.49 (d, J=19.4 Hz, 1H), 4.37 (s, 2H), 4.32-4.24 (m, 2H), 4.16 (d, J=19.4 Hz, 1H), 3.95-3.88 (m, 4H), 3.79 (d, J=5.5 Hz, 2H), 3.72 (d, J=5.8 Hz, 2H), 2.58-2.52 (m, 2H), 2.30-2.24 (m, 2H), 2.17-2.05 (m, 1H), 2.05-1.89 (m, 2H), 1.82-1.58 (m, 6H), 1.39 (s, 3H), 1.11-0.96 (m, 2H), 0.85 (s, 3H).

    ##STR01043##

    [0744] Step 1 Compound 5 (1.9 g, 3.21 mmol, 1.0 eq), Compound 8D (1.5 g, 3.21 mmol, 1.0 eq) and HATU (1.6 g, 4.24 mmol, 1.3 eq) were added into a 100-mL three-necked flask, and the reaction mixture was dissolved in DMF (45 mL), added slowly with 2,6-lutidine (1.4 g, 12.72 mmol, 4.0 eq). After the starting material was consumed completely as detected by LCMS, the reaction solution was added slowly and dropwise to water (300 mL) and filtered to give the product which was dried by oil pump under vacuum to give a yellow solid 10A (3.3 g, yield: 94%). MS-ESI: m/z 1048.4 [M+H].sup.+.

    [0745] Step 2 Compound 10A (1.8 g, 1.72 mmol, 1.0 eq) and Pd(PPh.sub.3).sub.4 (392 mg, 0.34 mmol, 0.2 eq) were added into a 50-mL three-necked flask, the reaction mixture was dissolved in dichloromethane (30 mL), under nitrogen atmosphere, added slowly with N-methylmorpholine (868 mg, 8.6 mmol, 5.0 eq) and reacted for 0.5 h. After the reaction was completed as detected by LCMS, diethylamine (6 mL) was added slowly into the three-necked flask and stirred at room temperature for 1 h. After the reaction was completed as detected by LCMS, the reaction solution was added with DMF (20 mL), subjected to rotary evaporation under vacuum at room temperature until 20 mL of the solvent remained, then the rotary evaporation was stopped, and the residue was subjected to reversed-phase column chromatography (ACN in water from 30%-70%) and lyophilized to give a light yellow solid 10B (455 mg, yield: 34%).

    [0746] Step 3 Compound 10B (150 mg, 0.19 mmol, 1.0 eq) was dissolved in THF (5 mL) and water (0.2 mL), triethylamine (190 mg, 1.9 mmol, 10 eq) was added, and the reaction solution was cooled to 0 C. in an ice-water bath. Bromoacetyl bromide (153 mg, 0.76 mmol, 4.0 eq) was dissolved in 1 mL THF, and then the resulting solution was added slowly to the above reaction solution and reacted at 0 C. for 15 min. After the reaction was completed as detected by LCMS, the reaction solution was subjected to preparative chromatography to give a white solid 10 (20 mg, yield: 12%).

    [0747] MS-ESI: m/z 906.3 [M+H].sup.+.

    [0748] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.87 (s, 1H), 8.52 (s, 1H), 8.22 (d, J=7.6 Hz, 1H), 7.43 (s, 1H), 7.38 (d, J=7.9 Hz, 2H), 7.31 (d, J=9.7 Hz, 2H), 7.23 (d, J=7.9 Hz, 2H), 6.87 (s, 1H), 6.16 (d, J=10.3 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=5.1 Hz, 1H), 4.78 (s, 1H), 4.49 (d, J=19.2 Hz, 1H), 4.41 (s, 2H), 4.39-4.32 (m, 1H), 4.29 (s, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.93 (s, 2H), 3.88 (s, 2H), 3.82-3.77 (m, 2H), 2.59-2.53 (m, 1H), 2.35-2.21 (m, 4H), 2.04-1.92 (m, 2H), 1.84-1.63 (m, 6H), 1.39 (s, 3H), 1.09-0.99 (m, 2H), 0.86 (s, 3H).

    ##STR01044## ##STR01045##

    [0749] Step 1 Compound 6 (300 mg, 0.5 mmol, 1.0 eq), Compound 8D (233 mg, 0.5 mmol, 1.0 eq) and HATU (228 mg, 0.6 mmol, 1.2 eq) were added into a 25-mL three-necked flask, and the reaction mixture was dissolved in DMF (5 mL), and added slowly with 2,6-lutidine (150 mg, 1.4 mmol, 2.8 eq). After the starting material was consumed completely as detected by LCMS, the reaction solution was added slowly and dropwise to water (100 mL) and filtered to give the product which was dried by oil pump under vacuum to give a yellow solid 11A (450 mg, yield: 86%). MS-ESI: m/z 1048.4 [M+H].sup.+.

    [0750] Step 2 Compound 11A (450 mg, 0.43 mmol, 1.0 eq) and Pd(PPh.sub.3).sub.4 (92 mg, 0.08 mmol, 0.2 eq) were added into a 50-mL three-necked flask, the reaction mixture was dissolved in THE (10 mL), and under nitrogen atmosphere, the reaction solution was added slowly with N-methylmorpholine (200 mg, 1.98 mmol, 4.6 eq) and reacted for 0.5 h. After the reaction was completed as detected by LCMS, the reaction solution was subjected to reversed-phase column chromatography (ACN in water from 30%-70%) and lyophilized to give a light yellow solid 11B (250 mg, yield: 58%). MS-ESI: m/z 1008.3 [M+H].sup.+.

    [0751] Step 3 Compound 11B (250 mg, 0.25 mmol) was added into a 25-mL three-necked flask, the reaction mixture was dissolved in DCM (10 mL), and under nitrogen atmosphere, the reaction solution was added with diethylamine (2 mL) and reacted for 0.5 h. After the reaction was completed as detected by LCMS, the reaction solution was subjected to reversed-phase column chromatography (ACN in water from 20%-50%) and lyophilized to give a light yellow solid 11C (130 mg, yield: 67%).

    [0752] MS-ESI: m/z 786.4 [M+H].sup.+.

    [0753] Step 4 Compound 11C (120 mg, 0.153 mmol, 1.0 eq) was dissolved in THE (5 mL) and water (0.2 mL), triethylamine (155 mg, 1.53 mmol, 10.0 eq) was added, and the reaction solution was cooled to 0 C. in an ice-water bath. Bromoacetyl bromide (123 mg, 0.61 mmol, 4.0 eq) was dissolved in 1 mL THF, and then the resulting solution was added slowly to the above reaction solution and reacted at 0 C. for 15 min. After the reaction was completed as detected by LCMS, the reaction solution was subjected to prep-HPLC to give a white solid 11 (12.2 mg, yield: 9%).

    [0754] MS-ESI: m/z 906.3 [M+H].sup.+.

    [0755] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.51 (s, 1H), 8.58-8.45 (m, 1H), 8.37 (d, J=7.6 Hz, 1H), 7.51-7.46 (m, 1H), 7.40-7.34 (m, 2H), 7.31 (d, J=10.0 Hz, 1H), 7.28-7.19 (m, 3H), 7.01-6.94 (m, 1H), 6.16 (dd, J=10.0, 1.9 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=5.0 Hz, 1H), 4.77 (brs, 1H), 4.49 (d, J=19.5 Hz, 1H), 4.42 (d, J=3.7 Hz, 1H), 4.39-4.32 (m, 1H), 4.29 (brs, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.98-3.77 (m, 6H), 2.60-2.52 (m, 2H), 2.34-2.28 (m, 2H), 2.18-1.96 (m, 3H), 1.88-1.55 (m, 6H), 1.39 (s, 3H), 1.13-0.98 (m, 2H), 0.85 (s, 3H).

    ##STR01046##

    [0756] Step 1 Compound 4C (10.0 g, 44.2 mmol) in DCM (70 mL) was added with TEA (8.94 g, 88.39 mmol, 12.30 mL), and the reaction solution was cooled to 0 C., added with MsCl (6.08 g, 53.0 mmol, 4.10 mL), and stirred at 25 C. for 12 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was diluted with DCM (50 mL), quenched with water (200 mL), and extracted with DCM (100 mL3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, and filtered. and the filtrate was concentrated by rotary evaporation to give Compound 12A (8.60 g, crude) as a yellow oil.

    [0757] Step 2 Compound 12A (8.60 g, 31.58 mmol) in DMF (51.6 mL) was added with thioglycolic acid (7.21 g, 63.15 mmol), and stirred at 25 C. for 5 h. After the starting material was consumed completely as detected by TLC (PE/EA=5:1), the reaction solution was added with water (100 mL), and extracted with ethyl acetate (100 mL3). The organic phases were combined, washed with saturated saline solution and then with 5% aqueous LiCl solution, dried over anhydrous Na.sub.2SO.sub.4, and filtered, and the filtrate was concentrated by rotary evaporation to give Compound 12B (5.75 g, yield: 57.82%, purity: 90.2%) as a brown oil. MS-ESI: m/z 285.2 [M+H].sup.+.

    [0758] Step 3 Compound 12B (12.0 g, 42.2 mmol) was dissolved in methanol (240 mL), and the reaction solution was added with K.sub.2CO.sub.3 (8.75 g, 63.3 mmol) and stirred at 20 C. for 3 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was poured into DCM (240 mL), and 0.5 M HCl (100 mL) was added, followed by liquid separation. The organic phase was dried over anhydrous sodium sulfate, and concentrated to give an off-white solid 12C (10.0 g, yield: 97.7%). MS-ESI: m/z 483.2 [M+H].sup.+.

    [0759] Step 4 Compound 12C (9.00 g, 37.1 mmol), Compound 1C (13.9 g, 37.1 mmol) and anhydrous magnesium sulfate (13.4 g, 111 mmol) were added to acetonitrile (270 mL), TfOH (16.7 g, 111 mmol, 9.84 mL) was added dropwise at 0 C., and the reaction solution was gradually heated to 20 C. after the addition was completed, and stirred for 3 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was quenched with saturated aqueous sodium bicarbonate (60 mL), and extracted with ethyl acetate (100 mL2). The organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and concentrated to give an off-white solid 12D (15.0 g, yield: 33.6%). MS-ESI: m/z 1199.4 [M+H].sup.+.

    [0760] Step 5 Compound 12D (290 mg, crude) was added to DMF (4 mL), then DL-dithiothreitol (200 mg, 1.30 mmol) was added, and the reaction solution was reacted at room temperature for 16 h under nitrogen atmosphere. After the reaction was completed as detected by LCMS, the reaction solution was directly subjected to prep-HPLC to give a white powdery solid 12 (20 mg, yield: 13.8%). MS-ESI: m/z 601.2 [M+H].sup.+.

    [0761] .sup.1H NMR (400 MHz, DMSO) 7.37 (d, J=8.1 Hz, 2H), 7.31 (d, J=10.1 Hz, 1H), 7.23 (dd, J=8.1, 3.3 Hz, 4H), 7.14 (d, J=8.0 Hz, 2H), 6.16 (dd, J=10.1, 1.7 Hz, 1H), 5.93 (s, 1H), 5.39 (s, 1H), 4.91 (d, J=5.0 Hz, 1H), 4.78 (s, 1H), 4.49 (d, J=19.4 Hz, 1H), 4.29 (s, 1H), 4.17 (d, J=19.5 Hz, 1H), 3.88 (d, J=11.3 Hz, 2H), 3.67 (d, J=7.6 Hz, 2H), 2.77 (t, J=7.6 Hz, 1H), 2.50-2.55 (m, 1H), 2.45-2.25 (m, 1H), 2.25-2.05 (m, 2H), 1.80-1.53 (m, 5H), 1.38 (s, 3H), 1.14-0.94 (m, 2H), 0.86 (s, 3H).

    ##STR01047## ##STR01048##

    [0762] Step 1 Compound 12 (7.00 g, 11.6 mmol) and Compound 8A (4.29 g, 11.6 mmol) were added to DMF (49.0 mL), and the reaction solution was added with TfOH (3.50 g, 23.3 mmol, 2.06 mL) and stirred at 25 C. for 16 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was quenched with saturated aqueous sodium bicarbonate (20 mL), and extracted with ethyl acetate (100 mL3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give Compound 13A as a yellow oil (15.0 g, crude product did not calculate yield). MS-ESI: m/z 909.3 [M+H].sup.+.

    [0763] Step 2 Compound 13A (15.0 g, 16.5 mmol) was added to acetonitrile (105 mL), and the reaction solution was added with diethylamine (6.03 g, 82.5 mmol, 8.50 mL) and stirred at 25 C. for 16 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was slurried with acetonitrile (20.0 mL) and filtered, the resulting filter cake was suctioned to dry, and the crude product was subjected to preparative reversed-phase chromatography (water:acetonitrile=25%-55%, 20 min), and lyophilized to give Compound 13B as a white solid (0.900 g, yield: 7.94%).

    [0764] MS-ESI: m/z 687.3 [M+H].sup.+.

    [0765] Step 3 Compound 13B (500 mg, 727 mol) and Compound 8D (526 mg, 1.09 mmol) were added to DMF (5 mL), and the reaction solution was added with HATU (830 mg, 2.18 mmol) and 2,6-lutidine (56.0 mg, 1.46 mmol, 169 L), and stirred at 20 C. for 3 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was dried under vacuum to give a crude product (550 mg), 250 mg of which was subjected to preparative reversed-phase chromatography to give Compound 13C (100 mg, yield: 26.2%) as a white solid. MS-ESI: m/z 1151.4 [M+H].sup.+.

    [0766] Step 4 Compound 13C (50.0 mg, 43.4 mol) was added to acetonitrile (1 mL), and the reaction solution was added with diethylamine (15.8 mg, 217 mol, 22.3 L) and stirred at 25 C. for 6 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was concentrated by rotary evaporation and slurried with methyl tert-butyl ether (3.00 mL) to give Compound 13D as a yellow solid (40.0 mg, yield: 99.1%). MS-ESI: m/z 929.2 [M+H].sup.+.

    [0767] Step 5 EEDQ (31.9 mg, 129 mol) and bromoacetic acid (11.3 mg, 81.8 mol) were dissolved in DMF (0.8 mL), and the reaction solution was stirred at room temperature for 1 h, added with Compound 13D (40.0 mg, 43.0 mol), and stirred at 25 C. for 2 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was concentrated by rotary evaporation to give Compound 13E as a yellow solid (the crude product was directly used in the next step). MS-ESI: m/z 1049.2 [M+H].sup.+.

    [0768] Step 6 Compound 13E (45.0 mg, 42.8 mol) was dissolved in DCM (1.50 mL), and the reaction solution was added with trifluoroacetic acid (770 mg, 6.75 mmol, 500 L) and stirred at 25 C. for 1 h. After the starting material was consumed completely as detected by LCMS, the reaction solution was concentrated by rotary evaporation, then the crude product was purified by preparative reversed-phase chromatography to give Compound 13 (3.06 mg, yield: 7.18%) as a yellow solid. MS-ESI: m/z 993.2 [M+H].sup.+.

    [0769] .sup.1HNMR (400 MHz, DMSO-d.sub.6) 7.83 (br d, 1H, J=7.6 Hz), 7.63 (br d, 1H, J=7.2 Hz), 7.4 (m, 2H), 7.26 (br d, 2H, J=8.0 Hz), 7.16 (br d, 1H, J=7.6 Hz), 4.92 (br s, 1H), 4.48 (br s, 1H), 4.30 (br s, 2H), 4.1-4.3 (m, 2H), 4.02 (br d, 1H, J=13.6 Hz), 3.92 (br d, 2H, J=8.4 Hz), 3.81 (br d, 2H, J=5.2 Hz), 3.74 (br s, 3H), 3.63 (br s, 2H), 2.91 (s, 2H), 2.75 (br s, 1H), 2.3-2.4 (m, 2H), 2.27 (br s, 2H), 2.1-2.1 (m, 1H), 2.02 (br s, 1H), 1.92 (br s, 1H), 1.77 (br s, 3H), 1.3-1.5 (m, 8H), 1.25 (s, 2H), 1.18 (br d, 1H, J=7.2 Hz), 1.02 (br s, 1H), 0.8-1.0 (m, 2H).

    ##STR01049##

    [0770] It was synthesized according to the reference patent CN109476699A.

    ##STR01050##

    [0771] It was synthesized according to the reference patent CN111465399A.

    ##STR01051##

    [0772] It was synthesized according to the reference patent CN111417410A.

    Example 2

    [0773] In the present application, an aqueous buffer of 20 mM histidine (pH=5.5) was obtained by adjusting the pH of an aqueous 20 mM Histidine solution to 5.5 with HOAc. Stock solution 1: 10 mM EDTA buffer, pH=4.7; stock solution 2: 800 mM sodium citrate aqueous buffer, pH=8.4; stock solution 3: 500 mM aqueous urea solution; stock solution 4: 10 mM aqueous tris(2-carboxyethyl)phosphine solution.

    [0774] The protein purity of the ADC in the present application was determined by SEC method, and the parameters are shown below.

    TABLE-US-00012 Instrument Agilent Technologies 1260 Chromatography TOSONG3000SWXL, 300 * 7.8 mm, 5 m column Column temperature 22 C. Wavelength 280 nm Injection volume 30~50 g Mobile phase 0.2M K.sub.2HPO.sub.4/KH.sub.2PO.sub.4, 0.25M KCl, 15%(v/v) 2-propanol, pH 7.0 Gradient Time (min) Flow rate (mL/min) 0.0 0.75 18.0 0.75

    [0775] The DAR value of the ADC in the present application can be detected by LC-MS method. The experimental procedure was as follows: to a solution containing 40.0 g ADC sample was added 75.0 L 8.0 mol/L guanidine hydrochloride (Gdn-HCl), 5.0 L 1.0 mol/L Tris-HCl, and 2.0 L 1 mol/L DTT. The solution was diluted to 100.0 L with ultrapure water, mixed well and incubated at 22 C. for 30 min. Drug/antibody ratio (DAR) was then analyzed using LC-MS. The LC parameters are shown below.

    TABLE-US-00013 Instrument Agilent Technologies 1260 Reversed-phase Agilent, PLRP-S 2.1*50 mm, 8 m chromatography column Column temperature 80 C. Sample temperature 2~8 C. Detection wavelength 280 nm Injection volume 10.0 L Mobile phase Mobile phase A: 0.025% TFA and 0.1% FA in ultrapure water Mobile phase B: 0.025% TFA and 0.1% FA in ACN Flow rate Time (min) % A % B (mL/min) Gradient 0.0 75 25 0.5 0.0 75 25 0.7 66 34 5.0 55 45 6.0 10 90 7.0 10 90 7.1 75 25 10.0 75 25

    [0776] The MS parameters are shown below.

    TABLE-US-00014 Instrument Agilent Technologies 6224 TOF LC/MS Ionization mode Positive Gas temperature 350 C. Air flow 13 L/min Atomizer pressure 45 psig Capillary voltage 5000 V Crushing voltage 250 V Voltage of taper hole 65 V Peak-to-peak value of radio 750 V frequency voltage on octupole Acquisition mode MS (seg) Mass range 500-8000 m/z Acquisition rate 1 spectra/s Acquisition time 1.4-7.0 min

    [0777] The DAR value of the ADC in the present application can also be detected by HIC method, and the parameters are shown below.

    TABLE-US-00015 Chromatography TSKgel Butyl-NPR, 4.6 mm *3.5 cm, 2.5 m column Column temperature 25 C. Detection 280 nm wavelength Sample load 8 L Mobile phase Mobile phase A: 1.5M (NH.sub.4).sub.2SO.sub.4, 0.05M K.sub.2HPO.sub.43H.sub.2O (pH = 7.0) Mobile phase B: 21.3 mM KH.sub.2PO.sub.4, 28.6 mM K.sub.2HPO.sub.43H.sub.2O, 25% (V/V) IPA (pH = 7.0) Flow rate Time (min) % B (mL/min) Gradient 0.0 0.0 0.6 2.0 0.0 15.0 100.0 16.0 100.0 17.0 0.0 20.0 0.0

    [0778] Murine anti-TNF mIgG2a 8c11: the light chain amino acid sequence is set forth as SEQ ID NO: 31 and the heavy chain amino acid sequence is set forth as SEQ ID NO: 32, prepared with reference to WO2015191783A2.

    [0779] Antibody Adalimumab (Humira): the light chain amino acid sequence is set forth as SEQ ID NO: 9 and the heavy chain amino acid sequence is set forth as SEQ ID NO: 10, prepared with reference to U.S. Pat. No. 6,090,382A.

    [0780] Antibody Litifilimab (BIIB059): the light chain amino acid sequence is set forth as SEQ ID NO: 42 and the heavy chain amino acid sequence is set forth as SEQ ID NO: 37, prepared with reference to CN105452295B.

    [0781] Antibody Iscalimab: the light chain amino acid sequence is set forth as SEQ ID NO: 19 and the heavy chain amino acid sequence is set forth as SEQ ID NO: 20, prepared with reference to CN105949314B.

    ##STR01052##

    [0782] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of anti-mouse TNF mIgG2a 8c11 was added prepared aqueous solution of tris(2-carboxyethyl) phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0783] Compound IV-1 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M PBS buffer at pH 6.5, containing 0.001 M EDTA) to give PBS buffer (5.0 mg/mL, 1.1 mL) of the exemplary product of formula (Va).

    [0784] The pH value of the PBS buffer of ADC was adjusted to 9 with 1 M Tris buffer, the resulting solution was incubated at 37 C. for 24 h, and the pH was adjusted to 6.5 with 1 mol/L citrate buffer to give Conjugate 1 (3.3 mg/mL, 5.5 mL). N.sup.a-I can be detected by LC-MS.

    ##STR01053##

    [0785] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0786] Compound IV-12 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate 2 (5.0 mg/mL, 1.1 mL).

    [0787] N.sup.a-I can be detected by LC-MS.

    ##STR01054##

    [0788] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0789] Compound IV-11 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate 3 (5.3 mg/mL, 1.5 mL).

    [0790] N.sup.a-I can be detected by LC-MS.

    ##STR01055##

    [0791] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0792] Compound IV-60 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate 4 (4.7 mg/mL, 2.2 mL).

    [0793] N.sup.a-I can be detected by LC-MS.

    ##STR01056##

    [0794] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0795] Compound IV-81 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate 5 (2.9 mg/mL, 4.3 mL).

    [0796] N.sup.a-I can be detected by LC-MS.

    ##STR01057##

    [0797] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0798] Compound IV-91 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate 6 (3.6 mg/mL, 5.5 mL).

    [0799] N.sup.a-I can be detected by LC-MS.

    ##STR01058##

    [0800] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody Adalimumab (Humira) was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0801] Compound IV-1 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M PBS buffer at pH 6.5, containing 0.001 M EDTA) to give PBS buffer (2.8 mg/mL, 4.7 mL) of the exemplary product of formula (Va).

    [0802] The pH value of the PBS buffer of ADC was adjusted to 9 with 1 M Tris buffer, the resulting solution was incubated at 37 C. for 24 h, and the pH was adjusted to 6.5 with 1 mol/L citrate buffer to give Conjugate V-6 (3.3 mg/mL, 5.5 mL). N.sup.a-I can be detected by LC-MS.

    ##STR01059##

    [0803] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody Humira was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0804] Compound IV-12 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate V-13 (3.5 mg/mL, 5.6 mL).

    [0805] N.sup.a-I can be detected by LC-MS.

    ##STR01060##

    [0806] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody Humira was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0807] Compound IV-11 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate V-12 (4.4 mg/mL, 4.3 mL).

    [0808] N.sup.a-I can be detected by LC-MS.

    ##STR01061##

    [0809] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody Humira was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0810] Compound IV-60 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate V-53 (2.7 mg/mL, 4.7 mL).

    [0811] N.sup.a-I can be detected by LC-MS.

    ##STR01062##

    [0812] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0813] Compound IV-81 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, and the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through SephadexG25 gel column (eluent: 0.05 M aqueous histidine buffer at pH 5.5, containing 0.001 M EDTA) to give Conjugate V-74 (2.8 mg/mL, 5.1 mL).

    [0814] N.sup.a-I can be detected by LC-MS.

    ##STR01063##

    [0815] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=7.4, containing 0.29 mM EDTA and 11.53 mM sodium citrate; 10.6 mL, 3.4 mg/mL, 0.243 mol) of antibody 8c11 were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.073 mL, 0.729 mol) and Compound 9 (3.56 mg, 3.645 mol) in DMSO (0.36 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped.

    [0816] The reaction solution was added with 1/3 volumes of 100 mM aqueous histidine buffer (pH=5.5), and desalted and purified through Zeba desalting spin column (eluent: 0.02 M aqueous histidine buffer at pH 5.5) (40K MWCO) to give Conjugate 9-1 (36.94 mg, 4.22 mg/mL, yield: 102%). N.sup.a-I was determined as 3.69 by RP.

    [0817] Conjugate 9-1 (36.94 mg, 4.22 mg/mL) was mixed with 1.55 mL of Conjugate 10 (3.38 mg/mL, 5.24 mg, RP-DAR 6.13) to give Conjugate 9 (39 mg, 4.22 mg/mL). N.sup.a-I was determined as 4.05 by RP.

    ##STR01064##

    [0818] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=7.4, containing 0.29 mM EDTA and 11.53 mM sodium citrate; 13.2 mL, 3.4 mg/mL, 0.303 mol) of antibody 8c11 were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.164 mL, 1.64 mol) and Compound 9 (4.44 mg, 4.545 mol) in DMSO (0.44 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped.

    [0819] The reaction solution was added with 1/3 volumes of 100 mM aqueous histidine buffer (pH=5.5), and desalted and purified (eluent: 0.02M aqueous histidine buffer at pH 5.5) using a Zeba desalting spin column (40K MWCO) to give Conjugate 10 (39 mg, 3.38 mg/mL, yield: 87%).

    [0820] N.sup.a-I was determined as 6.13 by RP.

    ##STR01065##

    [0821] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=5.5, containing 0.29 mM EDTA and 11.53 mM sodium citrate; 11.9 mL, 3.4 mg/mL, 0.273 mol) of antibody 8c11 were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.123 mL, 1.23 mol) and Compound 11 (3.72 mg, 4.10 mol) in DMSO (0.37 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped.

    [0822] The reaction solution was added with 1/3 volumes of 100 mM aqueous histidine buffer (pH=5.5), and desalted and purified (eluent: 0.02M aqueous histidine buffer at pH 5.5) using a Zeba desalting spin column (40K MWCO) to give Conjugate 11 (20 mg, 2.48 mg/mL, yield: 49%).

    [0823] N.sup.a-I was determined as 4.58 by RP.

    ##STR01066##

    [0824] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=5.5; 3.8 mL, 11.3 mg/mL, 0.290 mol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.103 mL, 1.03 mol) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped. Stock solution 1 and stock solution 2 were added such that the buffer system could contain 0.29 mM EDTA and 11.53 mM sodium citrate.

    [0825] Compound 10 (4.73 mg, 5.22 mol) was dissolved in 0.47 mL of DMSO, the resulting solution was added to the above solution, placed in a water bath shaker, and shaken and reacted at 22 C. for 9 h before the reaction was stopped. The reaction solution was added with 1/3 volumes of 800 mM aqueous sodium citrate buffer (pH=5.5), and desalted and purified (eluent: 0.02 M aqueous histidine buffer at pH 5.5) using a Zeba desalting spin column (40K MWCO) to give Conjugate 12 (35.39 mg, 2.17 mg/mL, yield: 82%).

    [0826] N.sup.a-I was determined as 4.21 by RP.

    ##STR01067##

    [0827] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=5.5, containing 0.29 mM EDTA and 11.53 mM sodium citrate; 2.5 mL, 3.4 mg/mL, 0.057 mol) of antibody 8c11 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.026 mL, 0.26 mol) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped.

    [0828] Compound 13 (1.02 mg, 1.026 mol) was dissolved in 0.10 mL of DMSO, and the resulting solution was added to the above solution, placed in a water bath shaker, and shaken and reacted at 22 C. for 6 h before the reaction was stopped. The reaction solution was desalted and purified (eluent phase: 0.02 M aqueous histidine buffer at pH 5.5) using a Zeba desalting spin column (40K MWCO) to give Conjugate 13 (5.8 mg, 1.72 mg/mL, yield: 68%).

    [0829] N.sup.a-I was determined as 3.76 by LC-MS.

    ##STR01068##

    [0830] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=5.5, containing 0.51 mM EDTA and 20.35 mM sodium citrate; 11.0 mL, 6.0 mg/mL, 0.446 mol) of antibody 8c11 were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.174 mL, 1.74 mol) and Compound 16 (6.40 mg, 6.69 mol) in DMSO (0.64 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 4.5 h before the reaction was stopped.

    [0831] The reaction solution was added with 1/3 volumes of 800 mM aqueous sodium citrate buffer (pH=5.5) and 10% volumes of aqueous sucrose solution (60%), and desalted and purified using a Zeba desalted centrifugal column (40K MWCO) (eluent: 0.05 M aqueous sodium citrate buffer at pH 6.0, containing 6% sucrose) to give Conjugate 14 (54 mg, 3.18 mg/mL, yield: 82%).

    [0832] N.sup.a-I was determined as 4.69 by RP.

    ##STR01069##

    [0833] To an aqueous PB buffer (0.04 M aqueous PB buffer at pH=6.97, containing 100 mM NaCl; 10.0 mL, 20 mg/mL, 1.351 mol) of antibody Adalimumab were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.338 mL, 3.378 mol) and Compound 9 (18.49 mg, 18.91 mol) in DMSO (1.85 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 3 h before the reaction was stopped.

    [0834] To an aqueous PB buffer (0.04 M aqueous PB buffer at pH=6.97, containing 100 mM NaCl; 60.0 mL, 20 mg/mL, 8.108 mol) of antibody Adalimumab were added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 2.189 mL, 21.892 mol) and Compound 9 (111.00 mg, 113.51 mol) in DMSO (11.10 mL) at 22 C. The resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 5 h before the reaction was stopped.

    [0835] The reaction solutions were combined, purified with UFDF (24 DV, 3 CV buffer+2 CV buffer (containing 5% DMA)+3 CV buffer+2 CV buffer (5% DMA)+3 CV buffer+6 CV buffer+5 CV buffer, the buffer being 0.02 M aqueous histidine buffer at pH 5.5), and filtered through a 0.22 M membrane to give Conjugate 17 (1390 mg, 18.47 mg/mL, yield: 99%).

    [0836] N.sup.a-I was determined as 3.66 by LC-MS. N.sup.a-I was determined as 3.83 by RP. N.sup.a-I was determined as 3.77 by HIC.

    ##STR01070##

    [0837] To an aqueous NaOAc buffer (0.05 M aqueous NaOAc buffer at pH=5.5; 7.5 mL, 4.0 mg/mL, 0.203 mol) of antibody Iscalimab was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.173 mL, 1.726 mol), Compound 9 (2.98 mg, 3.045 mol) was dissolved in DMSO (0.30 mL), and the resulting solution was placed in a water bath shaker, and shaken and reacted at 22 C. for 4.5 h before the reaction was stopped.

    [0838] The reaction solution was desalted and purified (eluent: 0.02 M aqueous histidine buffer at pH 5.5) using a Zeba desalting spin column (40K MWCO) to give Conjugate 18 (28 mg, 3.72 mg/mL, yield: 93%).

    [0839] N.sup.a-I was determined as 4.31 as detected by LC-MS.

    ##STR01071##

    [0840] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody BIIB059 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0841] Compound 9 (1.98 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through Sephadex G25 gel column (eluent: 0.05 M aqueous histidine buffer at pH=5.5, containing 0.001 M EDTA) to give Conjugate 19 (2.42 mg/mL, 3.31 mL, yield: 77%).

    [0842] N.sup.a-I was determined as 4.75 by LC-MS.

    ##STR01072##

    [0843] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody BIIB059 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a water bath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0844] Compound 16 (1.93 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through Sephadex G25 gel column (eluent: 0.05 M aqueous histidine buffer at pH=5.5, containing 0.001 M EDTA) to give Conjugate 20 (2.56 mg/mL, 3.52 mL, yield: 83%).

    [0845] N.sup.a-I was determined as 4.27 by LC-MS.

    ##STR01073##

    [0846] To an aqueous PB buffer (0.05 M PBS buffer at pH=6.5; 2.5 mL, 9.96 mg/mL, 0.168 nmol) of antibody BIIB059 was added prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37 C., and the reaction solution was placed in a waterbath shaker, and shaken and reacted at 37 C. for 3 h before the reaction was stopped; the reaction solution was cooled to 25 C. in a water bath, diluted to 5.0 mg/mL, and 2.0 mL of the solution was taken out and used in the next step.

    [0847] Compound 15 (1.77 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, the resulting solution was added to the above solution (2.0 mL), placed in a water bath shaker, and shaken and reacted at 25 C. for 3 h before the reaction was stopped. The reaction solution was desalted and purified through Sephadex G25 gel column (eluent: 0.05 M aqueous histidine buffer at pH=5.5, containing 0.001 M EDTA) to give Conjugate 21 (2.49 mg/mL, 3.53 mL, yield: 79%).

    [0848] N.sup.a-I was determined as 4.30 by LC-MS.

    Example 3: Measurement of Glucocorticoid Receptor Reporter Gene GRE Activity

    3.1 Measurement 1 of Glucocorticoid Receptor Reporter Gene GRE Activity

    1. Cell Culturing

    [0849] HEK293 cells were cultured in DMEM+10% FBS medium in a carbon dioxide incubator humidified at 37 C. at 5% carbon dioxide.

    2. Plasmid Transfection and Cell Plating

    [0850] HEK293 cells were trypsinized, and the HEK293 cell resuspension was adjusted to 200,000 cell/mL in medium (96 well plate: 100 L/well).

    [0851] Transfection reagent mixtures were prepared according to Tables A and B below, mixed well and left at room temperature for 20 minutes.

    TABLE-US-00016 Experimental transfection material Volume (L) pGL5 plasmid (100 ng/L) 25 pBIND-GR(100 ng/uL) 25 Opti-MEM medium 250.0 P3000 10.00 Opti-MEM medium 250.0 Lipofectamine 7.5

    TABLE-US-00017 Experimental transfection material Volume (L) PLvx-EF1a-hBDCA2-CMV- 50 hFcRI-IRES-puro(100 ng/uL) pGL5 plasmid (100 ng/L) 25 pBIND-GR(100 ng/uL) 25 Opti-MEM medium 250.0 P3000 10.00 Opti-MEM medium 250.0 Lipofectamine 7.5

    [0852] The prepared transfection mixtures were added to 10 mL of the cell density-adjusted resuspension, mixed well by turning the mixtures upside down, and seeded into a 96-well plate at a volume of 100 L/well.

    [0853] The 96-well plate was incubated in a humidified incubator at 37 C. and 5% carbon dioxide for 24 hours.

    3. Previous Treatment

    [0854] A 210 M dexamethasone (purchased from the Sphsine) in DMSO was prepared and serially diluted at 3.3 to obtain 10 concentrations. 5 L of the above dexamethasone in DMSO with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain dexamethasone dilutions with 10 concentrations.

    [0855] 2.1 M of solutions of the compounds of the present application or the compounds of the reference examples (diluted with culture medium) were prepared and serially diluted at 3.3 to obtain test compound dilutions with 10 concentrations.

    [0856] 5 L of dexamethasone dilution or test compound dilution was added into each well (100 L of cells had been added). 5 L of 10% DMSO medium was added into Min wells, and 5 L 21 M dexamethasone (dissolved in medium, 10% DMSO) was added into Max wells.

    [0857] The test plate was placed back in the incubator and incubated for 24 hours.

    4. Luciferase Assay

    [0858] The test plate was detected using the Dual-Glo Luciferase Assay System kit from Promega, and the fluorescence signal of Firefly Luciferase and the fluorescence signal of Renilla Luciferase were read using Enspire, respectively.

    5. Data Processing

    [0859] The final measurement value was a normalized value F/R obtained by dividing the Firefly Luciferase fluorescent signal by the Renilla Luciferase fluorescent signal.

    [0860] The data were copied and pasted into Excel, and the activation rate was obtained through an equation.

    [00001] Rate of activation ( % ) = Final measurement value - MinMean value MaxMean value - MinMean value 100 %

    [0861] The data were imported into MS Excel and subjected to curve fitting by using XLFit excel add-in version 5.4.0.8 to obtain an EC.sub.50 value.

    [00002] Y = Bottom + ( Top - Bottom ) / ( 1 - ( EC 50 / X ) ^ HillSlope Formula

    [0862] Y is the agonism and X is the concentration of the compound.

    [0863] Table 6 shows that the compounds of the present application exhibited stronger glucocorticoid receptor agonistic activity.

    TABLE-US-00018 TABLE 6 Measurement 1 of glucocorticoid receptor reporter gene GRE activity Compound No. EC50 (nM) Dexamethasone 2.11 Compound 14 0.19 (Reference Compound P1) Compound 4 0.04 Compound 4S 1.42

    3.2 Measurement 2 of Glucocorticoid Receptor Reporter Gene GRE Activity

    [0864] Example HEK293 GRE cells were plated at 50,000 cells/well in 50 L assay medium (RPMI, 1% CSFBS, 1% L-glutamine, 1% sodium pyruvate, and 1% MEAA) on a 96-well tissue culture treated white plate (Costar: 3917). The compounds of the present application were serially three-fold diluted in 100% DMSO at an initial concentration of 1 M for 9 times. 2 L of serially-diluted compounds were transferred into 248 L of assay medium in a secondary dilution plate (1:125 dilution) such that the small molecule compounds were further diluted in the assay medium. The cells were then treated with 25 L of 1:125 diluted GR agonist compound or medium alone at a final initial concentration of 266.7 nM (1:3) and incubated at 37 C. with 5% CO.sub.2 for 24 hours. After 24 hours of incubation, the cells were treated with 75 L of Dual-Glo luciferase assay system (Promega-E2920) for 10 minutes and analyzed for luminescence using TopCount or MicroBeta2 (PerkinElmer).

    [0865] Table 7 shows that the compounds of the present application exhibited significant glucocorticoid receptor agonistic activity.

    TABLE-US-00019 TABLE 7 Measurement 2 of glucocorticoid receptor reporter gene GRE activity Compound No. EC50 (nM) Compound 14 0.43 (Reference Compound P1) Compound 6 0.12

    Example 4: GR Luciferase Reporter Gene Elution Experiment

    Objective

    [0866] The compounds of the present application were evaluated for their ability to continue activation upon binding to GR.

    Procedures

    1. Cell Culture

    [0867] HEK293 cells were cultured in DMEM+10% FBS medium in a carbon dioxide incubator humidified at 37 C. at 5% carbon dioxide.

    2. Plasmid Transfection and Cell Plating

    [0868] 1) HEK293 cells were trypsinized, and the HEK293 cell resuspension was adjusted to 200,000 cell/mL in medium (96 well plate: 100 L/well). [0869] 2) Transfection reagent mixtures were prepared according to the table below, mixed well and left at room temperature for 20 minutes.

    TABLE-US-00020 Experimental transfection material Volume (L) pGL5 plasmid (100 ng/L) 25 pBIND-GR(100 ng/uL) 25 Opti-MEM medium 250.0 P3000 10.00 Opti-MEM medium 250.0 Lipofectamine 7.5 [0870] 3) The prepared transfection mixtures were added to 10 mL of the cell density-adjusted resuspension, mixed well by turning the mixtures upside down, and seeded into a 96-well plate at a volume of 100 L/well. [0871] 4) The 96-well plate was incubated in a humidified incubator at 37 C. and 5% carbon dioxide for 24 hours.

    3. Drug Treatment

    [0872] 1) 210 M dexamethasone in DMSO was prepared and serially diluted at 3 to obtain 10 concentrations. 5 L of the above dexamethasone in DMSO with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain dexamethasone dilutions with 10 concentrations. [0873] 2) A DMSO solution of the test compound at 210-fold final concentration was prepared and serially diluted at 3 to obtain dilutions of the test compound with 10 concentrations. 5 L of the above DMSO solution with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain dilutions with 10 concentrations. [0874] 3) 5 L of dexamethasone dilution or test compound dilution was added into each well (100 L of cells had been added). 5 L of 10% DMSO medium was added into Min wells, and 5 L 21 M dexamethasone (dissolved in medium, 10% DMSO) was added into Max wells. [0875] 4) After the test plate was placed back into the incubator for incubation for a certain period of time (15 min/30 min/1 h/2 h/4 h), the supernatant was carefully and slowly removed with a pipette and added into the medium at 100 L/well, then the supernatant was carefully and slowly removed again and finally added into the medium at 100 L/well, followed by placement into the incubator for incubation for 24 hours.

    4. Luciferase Assay

    [0876] The test plate was detected using the Dual-Glo Luciferase Assay System kit from Promega, and the fluorescence signal of Firefly Luciferase and the fluorescence signal of Renilla Luciferase were read using Enspire, respectively.

    5. Data Processing

    [0877] 1) The final measurement value was a normalized value F/R obtained by dividing the Firefly Luciferase fluorescent signal by the Renilla Luciferase fluorescent signal. [0878] 2) The data were copied and pasted into Excel, and the activation rate was obtained through an equation.

    [00003] Rate of activation ( % ) = Final measurement value - MinMean value MaxMean value - MinMean value 100 % [0879] 3) The data were imported into MS Excel and subjected to curve fitting by using XLFit excel add-in version 5.4.0.8 to obtain an EC.sub.50 value.

    [00004] Y = Bottom + ( Top - Bottom ) / ( 1 - ( EC 50 / X ) ^ HillSlope Formula

    [0880] Y is the agonism and X is the concentration of the compound.

    Experimental Results

    [0881] The result in Table 8 shows that the compounds of the present application bound more tightly to the glucocorticoid receptor, were less likely to be eluted, and had a longer lasting activation of the glucocorticoid receptor.

    TABLE-US-00021 TABLE 8 GR luciferase reporter gene elution experiment result Wash off time (Rate of activation %) Compound No. 15 min 30 min 1 h 2 h 4 h Compound 4 127.6 127.0 164.6 131.3 139.9 129.6 128.5 106.6 124.8 125.3 Compound 14 15.2 14.6 0.6 15.3 0.1 (Reference Compound P1)

    Example 5: MR Luciferase Reporter Gene Experiment

    1. Cell Culture

    [0882] HEK293 cells were cultured in DMEM+10% FBS medium in a carbon dioxide incubator humidified at 37 C. at 5% carbon dioxide.

    2. Plasmid Transfection and Cell Plating

    [0883] 1) HEK293 cells were trypsinized, and the HEK293 cell resuspension was adjusted to 200,000 cell/mL in medium (96 well plate: 100 L/well). [0884] 2) Transfection reagent mixtures were prepared according to the table below, mixed well and left at room temperature for 20 minutes.

    TABLE-US-00022 Experimental transfection material Volume (L) pGL5 plasmid (100 ng/L) 25 pBIND-MR(100 ng/uL) 25 Opti-MEM medium 250.0 P3000 10.00 Opti-MEM medium 250.0 Lipofectamine 7.5 [0885] 3) The prepared transfection mixtures were added to 10 mL of the cell density-adjusted resuspension, mixed well by turning the mixtures upside down, and seeded into a 96-well plate at a volume of 100 L/well. [0886] 4) The 96-well plate was incubated in a humidified incubator at 37 C. and 5% carbon dioxide for 24 hours.

    3. Drug Treatment

    [0887] 5) 21 M aldosterone in DMSO was prepared and serially diluted at 3.3 to obtain 10 concentrations. 5 L of the above aldosterone in DMSO with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain aldosterone dilutions with 10 concentrations. [0888] 6) A DMSO solution of the test compound at 210-fold final concentration was prepared and serially diluted at 3.3 to obtain dilutions of the test compound with 10 concentrations. 5 L of the above DMSO solution with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain dilutions with 10 concentrations. [0889] 7) 5 L of aldosterone dilution or test compound dilution was added into each well (100 L of cells had been added). 5 L of 10% DMSO medium was added into Min wells, and 5 L 2.1 M aldosterone (dissolved in medium, 10% DMSO) was added into Max wells. [0890] 8) The test plate was placed back in the incubator and incubated for 24 hours.

    4. Luciferase Assay

    [0891] The test plate was detected using the Dual-Glo Luciferase Assay System kit from Promega, and the fluorescence signal of Firefly Luciferase and the fluorescence signal of Renilla Luciferase were read using Enspire, respectively.

    5. Data Processing

    [0892] 4) The final measurement value was a normalized value F/R obtained by dividing the Firefly Luciferase fluorescent signal by the Renilla Luciferase fluorescent signal. [0893] 5) The data were copied and pasted into Excel, and the activation rate was obtained through an equation.

    [00005] Rate of activation ( % ) = Final measurement value - MinMean value MaxMean value - MinMean value 100 % [0894] 6) The data were imported into MS Excel and subjected to curve fitting by using XLFit excel add-in version 5.4.0.8 to obtain an EC.sub.50 value.

    [00006] Y = Bottom + ( Top - Bottom ) / ( 1 - ( EC 50 / X ) ^ HillSlope Formula

    [0895] Y is the agonism and X is the concentration of the compound.

    TABLE-US-00023 TABLE 9 MR luciferase reporter gene experiment result Compound ID MR EC50 (nM) Dexamethasone >1000.00 Compound 4 >1000.00 Compound 5 >1000.00 Compound 3 >1000.00

    [0896] According to the result in Table 9, the compounds of the present invention had a low affinity for mineralocorticoid receptors.

    Example 6: ER Luciferase Reporter Gene Experiment

    1. Cell Culture

    [0897] HEK293 cells were cultured in DMEM+10% FBS medium in a carbon dioxide incubator humidified at 37 C. at 5% carbon dioxide.

    2. Plasmid Transfection and Cell Plating

    [0898] 1) HEK293 cells were trypsinized, and the HEK293 cell resuspension was adjusted to 200,000 cell/mL in medium (96 well plate: 100 L/well). [0899] 2) Transfection reagent mixtures were prepared according to the table below, mixed well and left at room temperature for 20 minutes.

    TABLE-US-00024 Experimental transfection material Volume (L) pGL5 plasmid (100 ng/L) 25 pBIND-ER(100 ng/uL) 25 Opti-MEM medium 250.0 P3000 10.00 Opti-MEM medium 250.0 Lipofectamine 7.5 [0900] 3) The prepared transfection mixtures were added to 10 mL of the cell density-adjusted resuspension, mixed well by turning the mixtures upside down, and seeded into a 96-well plate at a volume of 100 L/well. [0901] 4) The 96-well plate was incubated in a humidified incubator at 37 C. and 5% carbon dioxide for 24 hours.

    3. Drug Treatment

    [0902] 5) 21 M estradiol in DMSO was prepared and serially diluted at 3 to obtain 10 concentrations. 5 L of the above solution of estradiol in DMSO with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain estradiol dilutions with 10 concentrations. [0903] 6) A DMSO solution of the test compound at 210-fold final concentration was prepared and serially diluted at 3 to obtain dilutions of the test compound with 10 concentrations. 5 L of the above DMSO solution with each concentration was added into 45 L of DMEM+10% FBS respectively and mixed evenly to obtain dilutions with 10 concentrations. [0904] 7) 5 L of estradiol dilution or test compound dilution was added into each well (100 L of cells had been added). 5 L of 10% DMSO medium was added into Min wells, and 5 L 2.1 M estradiol (dissolved in medium, 10% DMSO) was added into Max wells. [0905] 8) The test plate was placed back in the incubator and incubated for 24 hours.

    4. Luciferase Assay

    [0906] The test plate was detected using the Dual-Glo Luciferase Assay System kit from Promega, and the fluorescence signal of Firefly Luciferase and the fluorescence signal of Renilla Luciferase were read using Enspire, respectively.

    5. Data Processing

    [0907] 9) The final measurement value was a normalized value F/R obtained by dividing the Firefly Luciferase fluorescent signal by the Renilla Luciferase fluorescent signal. [0908] 10) The data were copied and pasted into Excel, and the activation rate was obtained through an equation.

    [00007] Rate of activation ( % ) = Final measurement value - MinMean value MaxMean value - MinMean value 100 % [0909] 11) The data were imported into MS Excel and subjected to curve fitting by using XLFit excel add-in version 5.4.0.8 to obtain an EC.sub.50 value.

    [00008] Y = Bottom + ( Top - Bottom ) / ( 1 - ( EC 50 / X ) ^ HillSlope Formula

    Y is the agonism and X is the concentration of the compound.

    TABLE-US-00025 TABLE 10 ER luciferase reporter gene experiment result Compound ID ER EC50 (nM) Dexamethasone >1000.00 Compound 4 >1000.00 Compound 5 >1000.00 Compound 3 >1000.00

    [0910] According to the result in Table 10, the compounds of the present invention had a low affinity for estrogen receptors.

    Example 7: Inhibitory Effect of Compounds of the Present Invention on R848-Induced Release of IFN and TNF from Human Peripheral Blood Monocytes

    Objective

    [0911] The compounds of the present application were tested for their inhibitory effect on the production of peripheral blood monocytes. The peripheral blood monocytes were treated in vitro with the ligand-drug conjugate at different concentrations, and IFN and TNF generated by the peripheral blood monocytes were quantitatively detected after a certain time. The compounds of the present invention were evaluated for in-vitro activity.

    Test Method

    [0912] The cryopreserved human PBMC cells were resuspended in a complete medium and counted, and the cell suspension was adjusted to 2.510*6 cells/mL with the culture medium. After the suspension was centrifuged to collect the cells the next day, the cells were resuspended using fresh culture medium. Test compounds were diluted to 4-fold final concentration in cell culture medium, 50 L of the compounds were added into a 96-well cell plate, and 100 L of diluted cells (800,000 cells/well) were added into each well of the 96-well plate containing the compounds, followed by addition of 50 L R848 to a final concentration of 5 M. The cell plate was incubated at 37 C. with 5% CO.sub.2 for 24 hours. The supernatant was collected, and the TNF- and IFN- concentration levels in the supernatant were detected using an ELISA kit.

    [0913] TNF- concentration assay was performed using Dakewe TNF- ELISA kit (Cat #1117202), 100 L of the serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 10-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0914] IFN- concentration assay was performed using Dakewe TNF- ELISA kit (Cat #1110012), 100 L of the serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 2-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0915] The contents of TNF- and IFN- were calculated from the OD450 values. The results are shown in Table 11-1 and Table 11-2.

    TABLE-US-00026 TABLE 11-1 Inhibition of compounds on TNF- secretion (pg/mL) from PBMC under R848 stimulation Concentration Compound 14 of the (Reference Dexa- compound Compound 4 Compound P1) methasone Blank 100 nM 356.1 471.9 396.4 2507.3 0.2 138.4 88 25 20 nM 320 465.6 761.9 18.8 37.9 112.6 4 nM 443.6 868 1446.5 41.2 69.3 10.8 TNF- content was expressed as mean standard deviation.

    TABLE-US-00027 TABLE 11-2 Inhibition of compounds on IFN- secretion (pg/mL) from PBMC under R848 stimulation Concentration Compound 14 of the Compound Compound (Reference Dexa- compound 4 6 Compound P1) methasone Blank 100 nM 0 0 6.9 6.9 13 13 0 0 995.1 69.3 20 nM 27.9 27.9 45.9 45.9 10.2 10.2 169.6 94.4 4 nM 13.2 13.2 99.8 +5.7 146.4 16.6 620.7 70.4 IFN- content was expressed as mean standard deviation.

    [0916] The results show that the compounds of the present application could affect the ability of peripheral blood monocytes to produce TNF- and IFN-. The ligand-drug conjugates of the present application can inhibit inflammation, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    Example 8: Inhibitory Effect of Drug Conjugates of the Present Invention on R848-Induced Release of IFN, TNF, IL-6 and IL-8 from Human Peripheral Blood Monocytes

    Objective

    [0917] The inhibitory effect of the ligand-drug conjugate (ADC) of the present application on the production of IFN by peripheral blood monocytes was examined. The peripheral blood monocytes were treated in vitro with the ligand-drug conjugate at different concentrations, and IFN and TNF generated by the peripheral blood monocytes were quantitatively detected after a certain time. The ligand-drug conjugates were evaluated for in-vitro activity.

    Test Method

    [0918] The cryopreserved human PBMC cells were resuspended in a complete medium and counted, and the cell suspension was adjusted to 2.510*6 cells/mL with the culture medium. After the suspension was centrifuged to collect the cells the next day, the cells were resuspended using fresh culture medium, and 100 L of diluted cells (800,000 cells/well) were added into a 96-well plate. The test samples were diluted to 3-fold final stimulation concentration using fresh culture medium, 50 L of the diluted test samples were added into the 96-well cell plate, and the cell plate was incubated at 37 C. with 5% CO.sub.2 for 12 hours, followed by addition of 50 L 20 M R848 each well. The cell plate was incubated at 37 C. with 5% CO.sub.2 for 20 hours. The supernatant was collected, and the cytokine concentration in the supernatant was detected using a kit.

    [0919] TNF- concentration assay was performed using Dakewe TNF- ELISA kit (Cat #1117202), 100 L of the serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 10-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0920] IFN- concentration assay was performed using Dakewe TNF- ELISA kit (Cat #1110012), 100 L of the serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 2-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0921] IL-6 concentration assay was performed using Dakewe IL-6 ELISA kit (Cat #DKW12-1060-096), 100 L of serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 2-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0922] IL-8 concentration assay was performed using Dakewe IL-8 ELISA kit (Cat #1110802), 100 L of the serially-diluted cytokine standard was added into each well, 100 L of the above supernatant diluted 2-fold was added into each well, and 100 L of 1 dilution buffer R was added into each blank well. 50 L of biotinylated antibody working solution was added into each well. After mixing well, the plate was covered with the plate-sealing membrane and incubated at room temperature for 3 hours. The plate was washed for 3 times: the liquid in each well was discarded, and 300 L of 1 washing buffer working solution was added into each well; after 1 minute of residence, the liquid in the wells was discarded. The procedure was repeated for 3 times. 100 L of Streptavidin-HRP working solution was added into each well. The plate was covered with the plate-sealing membrane and incubated at room temperature for 20 minutes. The plate was washed for 3 times, 100 L of TMB liquid was added into each well, and the plate was incubated for 25 minutes at room temperature (18-25 C.) in the dark. The reaction was stopped by adding 100 L stop solution rapidly to each well. The reading at a wavelength of 450 nm was detected by an M2e instrument within 10 minutes after termination.

    [0923] The contents of TNF-, IFN-, IL-6 and IL-8 were calculated from the OD450 values, and the results are shown in FIGS. 1a, 1b, 1c and 1d.

    [0924] According to FIG. 1a, when the incubation concentration of antibodies BIIB059, Conjugate 19, Conjugate 20 and Conjugate 21 was 300 nm, the IFN- contents in the detection system were 1639.9 pg/mL, 113.2 pg/mL, 1422.5 pg/mL, 911.9 pg/mL, respectively; the inhibitory effect of Conjugate 19, Conjugate 20 and Conjugate 21 on IFN was better than that of antibody BIIB059, and the inhibitory effect of Conjugate 19 was significantly better than that of Conjugate 20 and Conjugate 21.

    [0925] According to FIG. 1b, when the incubation concentration of antibodies BIIB059, Conjugate 19, Conjugate 20 and Conjugate 21 was 300 nm, the IFN- contents in the detection system were 7040.1 pg/mL, 761.8 pg/mL, 3280.0 pg/mL, 2644.1 pg/mL, respectively; the inhibitory effect of Conjugate 19, Conjugate 20 and Conjugate 21 on IFN- was better than that of antibody BIIB059, and the inhibitory effect of Conjugate 19 was significantly better than that of Conjugate 20 and Conjugate 21.

    [0926] According to FIG. 1c, when the incubation concentration of antibodies BIIB059, Conjugate 19, Conjugate 20 and Conjugate 21 was 300 nm, the IL-6 contents in the detection system were 65932.1 pg/mL, 8861.7 pg/mL, 34744.1 pg/mL, 29867.1 pg/mL, respectively; the inhibitory effect of Conjugate 19, Conjugate 20 and Conjugate 21 on IL-6 was better than that of antibody BIIB059, and the inhibitory effect of Conjugate 19 was significantly better than that of Conjugate 20 and Conjugate 21.

    [0927] According to FIG. 1d, when the incubation concentration of antibodies BIIB059, Conjugate 19, Conjugate 20 and Conjugate 21 was 300 nm, the IL-8 contents in the detection system were 86905.7 pg/mL, 26565.5 pg/mL, 56315.8 pg/mL, 43544.9 pg/mL, respectively; the inhibitory effect of Conjugate 19, Conjugate 20 and Conjugate 21 on IL-8 was better than that of antibody BIIB059, and the inhibitory effect of Conjugate 19 was significantly better than that of Conjugate 20 and Conjugate 21.

    [0928] The results show that the ligand-drug conjugates of the present application could affect the ability of peripheral blood monocytes to produce IIFN, TNF, IL-6 and IL-8. The ligand-drug conjugates of the present application can inhibit inflammation, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    Example 9: Determination of Biological Activity in Fluorescein Isothiocyanate (FITC)-Induced Delayed-Type IV Hypersensitivity Mouse Model

    [0929] ADC was evaluated in an acute contact hypersensitivity model in which a delayed type hypersensitivity (DTH) response (T cell drive) was used to trigger acute skin inflammation by the application of sensitizers (fluorescein isothiocyanate, FITC). The potency of the ligand-drug conjugates (ADCs) of the present application was measured by their ability to reduce ear swelling. Before the experiment began, a specified number of 7-9 week-old female Balb/c mice were selected, random grouping was performed according to the weight of mice, with 10 mice in each group, and the average weight values among different groups were ensured to be similar so as to reduce the difference among the groups. The day of grouping the mice was defined as day 0 of the experiment. After random grouping, all mice were shaved off abdominal hair under mild anesthesia with 5% isoflurane, and mice in the disease-induced group were sensitized firstly by applying 100 L of 2% fluorescein isothiocyanate (FITC) solution to their abdomen, while mice in the disease-controlled group were applied with 100 L of vehicle control solution (acetone:DBP, volume ratio 1:1) to their abdomen. After the solvent solution and the 2% FITC solution were absorbed and the abdominal skin was dry, the mice were placed back into the cage box after they were awake. On day 1 of the experiment, the procedure described above was repeated for a second 2% FITC sensitization.

    [0930] All mice were weighed on day 5 of the experiment. According to the experimental design, the mice in the solvent control group or each treatment group were respectively injected with the sterile PBS solution or the specified administration solution in the abdominal cavity. 1 hour after dosing, all mice were tested for left and right ear thickness using Mitutoya Caliper under mild anesthesia with 5% isoflurane. After ear thickness measurement, all mice in the disease-induced group were coated with 10 L of 1% FTIC solution on the right ear and 10 L of vehicle control solution (acetone:DBP, volume ratio 1:1) on the left ear.

    [0931] On day 6 of the experiment, 24 hours after 1% FTIC excitation, all mice were tested for left and right ear thickness using Mitutoya Caliper under mild anesthesia with 5% isoflurane and mouse ear thickness change was calculated according to the following formula: (RL)(R0L0), wherein R0 and L0 are right and left ear thickness before 1% FITC excitation, respectively, and R and L are right and left ear thickness after 24 hours of 1% FITC excitation, respectively. The results are shown in FIG. 2. As shown in FIG. 2, when the administration concentration of Conjugate 9, antibody 8c11 and Conjugate 14 was 20 mg/kg, the mean increase in ear thickness of mice was 0.066 mm, 0.249 mm and 0.128 mm, respectively (see ADC1). The inhibitory effect of Conjugate 9 on hypersensitivity was significantly better than that of antibody 8c11 and Conjugate 14.

    [0932] The results show that the ligand-drug conjugate (ADC) of the present application had the ability to inhibit hypersensitivity, and that the ligand drug conjugate (ADC) of the present application could be used for preventing and/or treating diseases and or conditions of inflammation.

    Example 10: Determination of Bioactivity of Bovine Type II Collagen Mixed Adjuvant-Induced DBA/1 Mouse Arthritis Model

    [0933] The potency of the ligand-drug conjugates (ADCs) of the present application was evaluated in a collagen-induced arthritis (CIA) model.

    [0934] Male DBA/1J mice were obtained from Jackson Labs. The mice were used at 6-8 weeks of age. All animals were fed free diet water under 12 hours light/dark cycle at constant temperature and humidity. Body weight and condition were monitored and if >20% weight loss was exhibited, the animals were euthanized.

    [0935] Bovine collagen (Chondrex, Inc. Cat: 20021) was dissolved in 0.1 M acetic acid overnight in a 2-8 C. refrigerator at a collagen concentration of 8 mg/mL, and an equal volume of CFA (complete Freund's adjuvant, Difco, Laurence, KS) was added into the collagen solution and emulsified in a high-speed homogenizer to make an emulsion. At Day 0, animals in need of immunization were anesthetized with 2%-5% isoflurane and male DBA/J mice were immunized once with a single subcutaneous injection of 50 L collagen/CFA emulsion at the tail root of 2-3 cm from the body. Three weeks later, namely at Day 21, the tail root was immunized twice with the same volume of collagen emulsion by the same injection method. After the second immunization, 120 animals with an average score of about 2 for the limb arthritis were selected and randomly grouped. The animals were treated by a single intraperitoneal injection of test reagent according to the experimental design after grouping. For the dexamethasone (Sphsine) treatment group, the drug was administered the day after grouping until the end of the trial. The extremities of each group were observed twice weekly for the onset of arthritis from the time of the second immunization, and the observation was continued until the end of the trial.

    [0936] Scoring according to the degree of lesion (redness and swelling) was performed on a scale of 0-4 points, with a maximum score of 4 for each limb and a total maximum score of 16 for each animal's limbs. The scoring criteria are as follows: 0 points, no redness and swelling; 1 point, 1-2 red and swollen interphalangeal joints; 2 points, 3-4 red and swollen interphalangeal joints; 3 points, more than 4 red and swollen interphalangeal joints; 4 points, severe redness and swelling in toes or fingers to ankle joints or wrist joints. For joint swelling, ankle thickness of hind paw of each animal was measured with a screw micrometer before one immunization. Since the second immunization, measurements were performed 2 times per week, and changes in ankle thickness before and after the immunization were compared. After the trial was completed, all animals were anesthetized with 2-5% isoflurane and then euthanized by exsanguination.

    [0937] Score AUC: area under the curve for arthritis score.

    [0938] The results are shown in FIGS. 3a and 3b.

    [0939] The results show that the ligand-drug conjugates (ADCs) of the present application had the ability to reduce swelling of the hind paw of the mice, and could exhibit an extended duration of action of about 28 days, as compared to the blank agent, and had the ability to reduce the severity of arthritis and biological safety (e.g., blood safety).

    Example 11

    11.1 GRE Activity Assay

    Generation of Human and Mouse Transmembrane TNF- GRE Reporter Cell Lines

    [0940] To generate the parental cell line, K562 cells were seeded at 500,000 cells per well onto a 6-well dish (Costar: 3516) with 2 mL of complete growth medium (RPMI, 10% FBS, 1% L-glutamine, 1% sodium pyruvate, and 1% MEMNEAA (an optional amino acid solution)) at 37 C. with 5% CO.sub.2 within 24 hours. The next day, 1.5 g of pGL4.36 [Luc2P/MMTV/Hygro] (Promega) and 3 L of PLUS reagent (Invitrogen) were diluted into 244 L Opti-MEM (Gibco: 31985-070) and incubated at room temperature for 15 minutes. pGL4.36 [luc2P/MMTV/Hygro] vector contains the murine mammary tumor virus long terminal repeated sequence, which drives transcription of the luciferase reporter gene luc2P in response to activation of several nuclear receptors (e.g. glucocorticoid receptor and androgen receptor). After incubation, the diluted DNA solution was pre-incubated with 1:1 lipofectamine LTX solution (13.2 L+256.8 L Opti-MEM) and incubated at room temperature for 25 minutes to form a DNA-lipofectamine LTX complex. After incubation, 500 L of DNA-lipofectamine complex was added directly into the wells containing cells. K562 cells were transfected at 37 C. with 5% CO.sub.2 for 24 hours. After incubation, the cells were washed with 3 mL of PBS and selected for two weeks with complete growth medium containing 125 g/mL hygromycin B. K562pGL4.36 [Luc2P/MMTV/Hygro]_pGL4.75 [hRLuc/CMV] cells were generated.

    [0941] To generate murine transmembrane TNF- GRE reporter cells, the parent cells, K.sub.562pGL4.36 [Luc2P/MMTV/Hygro]_pGL4.75 [hRLuc/CMV], were seeded at 500,000 cells per well onto a 6-well dish containing 2 mL of complete growth medium (RPMI, 10% FBS, 1% L-glutamine, 1% sodium pyruvate and 1% MEMNEAA) at 37 C. with 5% CO.sub.2 within 24 hours. The next day, 3 g of mFL_TNF DNA encoding unlabeled mouse TNF and 3 L of PLUS reagent (Invitrogen: 10964-021) were diluted into 244 L of Opti-MEM (Gibco: 31985-070) and incubated at room temperature for 15 minutes. After incubation, the diluted DNA solution was pre-incubated with 1:1 lipofectamine LTX solution (13.2 L+256.8 L Opti-MEM) and incubated at room temperature for 25 minutes to form a DNA-lipofectamine LTX complex. After incubation, 500 L of DNA-lipofectamine complex was added directly into the wells containing cells. The parent K562pGL4.36\[Luc2P/MMTV/Hygro]_pGL4.75[hRLuc/CMV] cells were transfected at 37 C. with 5% CO.sub.2 for 24 hours. After incubation, the cells were washed with 3 mL of PBS and selected for two weeks with complete growth medium containing 125 g/mL hygromycin B (Invitrogen: 10687-010) and 250 g/mL G418 (Gibco: 10131-027). K562 mouse FL-TNFGRE (pGL4.36[luc2P/MMTV/Hygro]) cells were generated.

    [0942] To generate the human transmembrane TNF- GRE reporter cell line, the parent cell K562pGL4.36[Luc2P/MMTV/Hygro]_pGL4.75[hRLuc/CMV] was transfected with the hTNF1-12C-MycpcDNA3.1() plasmid construct. The plasmid was pcDNA3.1 (Thermo Fisher, Cat. V79020), which encodes tace-resistant transmembrane TNF. (See Perez C et al., Cell 63(2): 251-8 (1990) which discusses tace-resistant transmembrane TNF). K562 human TNF1-12GRE (pGL4.36[luc2P/MMTV/Hygro]) cells were generated. These cell lines were then used in the TNF- reporter assay described in the examples below.

    [0943] K562 parent GRE cells (K562pGL4.36 [Luc2P/MMTV/Hygro]_pGL4.75[hRLuc/CMV] cells), and GRE-mouse TNF- cells (K562 mouse FL-TNF- GRE(pGL4.36[luc2P/MMTV/Hygro]) cells) or GRE-human TNF- cells (K562 human TNF1-12GRE (pGL4.36[luc2P/MMTV/Hygro]) cells) were seeded onto a 96-well plate at 50,000 cells per well. The ligand-drug conjugates (ADCs) of the present application were serially diluted at 3 in media and added into the above 96-well plate and incubated at 37 C. with 5% CO.sub.2 for 48 hours. Luminescence was analyzed after treatment with the luciferase assay system. The data were analyzed using a four-parameter curve fit to generate EC.sub.50 values. The maximum % activation was normalized to 100 nM dexamethasone. The ligand-drug conjugates (ADCs) of the present application measured in-vitro activity in a mouse transmembrane TNF GRE reporter assay; and the ligand drug conjugates (ADCs) of the present application measured in-vitro activity in the human transmembrane TNF GRE reporter assay.

    [0944] The results show that the ligand-drug conjugates (ADCs) of the present application had the ability to affect the level of cellular GRE activation, and that the ligand drug conjugates (ADCs) of the present application could affect the level of glucocorticoid-mediated signaling pathway activation.

    11.2 Lipopolysaccharide (LPS) Induced Cytokine Release and Downstream Signal Detection

    [0945] Primary human peripheral blood monocytes (PBMCs) were washed in 50 mL PBS, resuspended in FBS (fetal bovine serum) with 5% DMSO, divided in equal and cryopreserved in liquid nitrogen until use. The PBMCs were thawed, resuspended in cell culture medium (e.g., RPMI cell culture medium) with 2% FBS and 1% penicillin streptomycin, and seeded onto a 96-well plate. The cells were then incubated with different concentrations of ADC for 4 hours at 37 C. with 5% CO.sub.2. The cells were treated with a stimulant such as lipopolysaccharides (LPS) at a certain concentration for a certain time. The plate was then spun at 1000 rpm for 5 minutes and 100 L of the supernatant culture medium was transferred directly into another 96-well plate, and the concentrations of cytokines such as IL-6 and IL-1 and phosphorylated STAT1 levels were analyzed.

    [0946] The results show that the ligand-drug conjugates (ADC) of the present application had the ability to affect the release of cytokines, such as IL-6 and IL-1, from PBMC cells, and the ability to phosphorylate STAT1.

    Example 12

    12.1 Type I Interferon-Induced Response Element Signal Detection

    [0947] HEK293 cells were constructed and transferred into pHTS-ISRE fluorescein reporter gene plasmid, and a reporter system of type I interferon response signals were constructed. The cells were cultured in a DMEM medium containing FBS and Geneticin with certain concentrations, and the serially-diluted ligand-drug conjugate and type I interferon IFN or recombinant type I interferon with certain concentrations induced by plasmacytoid dendritic cells were added for incubation. The cells were lysed, fluorescein intensity was determined, dose-response data were fitted to an S-shaped curve using non-linear regression, and IC.sub.50 values were calculated.

    [0948] The results show that the ligand-drug conjugates of the present application could influence type I interferon-induced response element signals. The ligand-drug conjugates of the present application can inhibit inflammation, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    12.2 Cytokine Release Induced by CpG-A and the Like and Downstream Signal Molecule Detection

    [0949] Primary human peripheral blood monocytes (PBMCs) were washed in 50 mL PBS, resuspended in FBS (fetal bovine serum) with 5% DMSO, divided in equal and cryopreserved in liquid nitrogen until use. The PBMCs were thawed, resuspended in cell culture medium (e.g., RPMI cell culture medium) with 2% FBS and 1% penicillin streptomycin, and seeded onto a 96-well plate. The cells were then incubated with different concentrations of the ligand-drug conjugate at 37 C. with 5% CO.sub.2. The cells were treated with CpG-A or other stimulants at a certain concentration for a certain time. The plate was then spun at 1000 rpm for 5 minutes and 100 L of the supernatant culture medium was transferred directly into another 96-well plate, and the levels of cytokines such as IL-6 and IL-1 were analyzed.

    [0950] The ligand-drug conjugate with a certain concentration was added into human peripheral blood monocytes for culturing. Then type I interferon IFN or recombinant type I interferon with certain concentrations induced by plasmacytoid dendritic cells was added for incubation. The lysed cells were subjected to electrophoresis and western blot detection, and STAT1 phosphorylation levels were determined using anti-human STAT1 pTY701 antibody.

    [0951] The results show that the ligand-drug conjugates (ADC) of the present application had the ability to affect the release of cytokines from PBMC cells, and the ability to phosphorylate STAT1.

    12.3 Evaluation of Drug Efficacy in Plasmacytoid Dendritic Cell Xenograft Mouse Model

    Objective

    [0952] The ligand-drug conjugates (ADCs) of the present application act on a human plasmacytoid dendritic cell xenograft mouse model, and the in-vivo drug effect of the ligand-drug conjugates is evaluated by means of immunohistochemistry, gene transcription analysis and the like.

    Test Method

    1. Imiquimod Induction Model

    [0953] The backs of 4-8 week-old severely-immunodeficient mice (CB17/Icr-Prkdcscid/IcrIcoCrl, Charles River) were shaved. The 5% imiquimod cream was applied to the backs of the mice, and 12 hours later, a second application was made. The mice were then injected intraperitoneally with a certain concentration of the ligand-drug conjugates of the present application or the control ligand-drug conjugate. 12 hours later, 1-1010.sup.5 human plasmacytoid dendritic cells were injected into the tail vein of the mice. After 12 hours of incubation, the mice were euthanized and dorsal skin samples were collected for testing.

    2. Bleomycin Induction Model

    [0954] The backs of 4-8 week-old severely-immunodeficient mice (CB17/Icr-Prkdcscid/IcrIcoCrl, Charles River) were shaved. Bleomycin with a certain concentration was injected subcutaneously into a single site on the backs of the mice for three weeks every two days. 1-1010.sup.5 human plasmacytoid dendritic cells were injected into the tail vein of the mice on Days 0, 7 and 14 of the first injection of bleomycin. The mice were injected intraperitoneally with the ligand-drug conjugates of the present application or the control ligand-drug conjugate with a certain concentration 24 hours before the first injection of bleomycin, and the injection was performed once every 5 days.

    3. Detection Method and Index

    [0955] (1) In the kit, mouse skin cell RNA was extracted and subjected to reverse transcription to obtain cDNA, and the influence of the ligand-drug conjugate on the transcription of the type I IFN signaling pathway response gene was determined by utilizing Real-time PCR and taking a mouse sample treated with a control ligand-drug conjugate as a control. [0956] (2) Mouse skin samples were fixed with formalin and embedded in paraffin, which was then sectioned at 5 M for hematoxylin-eosin staining. The degree of fibrosis of the skin samples was identified by Masson staining. Immunohistochemical analysis was performed with the pSTAT1 Tyr701 antibody. [0957] (3) After being processed, the mouse skin samples were analyzed by FACS, and the proportion of plasmacytoid dendritic cells in the mouse skin samples was determined according to the cell surface markers. [0958] (4) The collagen content of the mouse skin samples was measured by a collagen analysis method after being processed.

    [0959] The results show that the ligand-drug conjugates of the present application could affect (1) the ability of the type I IFN signaling pathway to respond to gene transcription, (2) the degree of fibrosis in a skin sample, (3) the ability of plasmacytoid dendritic cells to proliferate and/or (4) the collagen content of skin samples. The ligand-drug conjugate of the present application can inhibit inflammation in vivo, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    Example 13

    13.1 Cytokine Release Experiment for CD40L and LPS-Induced Dendritic Cells Derived from Human Peripheral Blood Monocytes

    [0960] Fresh human peripheral blood monocytes (PBMCs) were prepared. Human monocytes were isolated using a human monocyte (CD14+) isolation kit. After washing, the cells were resuspended, added with human IL-4 (80 ng/mL) and human GM-CSF (100 ng/mL) and placed in an incubator and cultured for 5 days.

    [0961] The cells were harvested, washed, added into a 96-well flat-bottom plate at 105 cell/well, and stimulated with 0.1 ng/mL LPS for 2 hours, and then the supernatant was washed off. A positive compound with a certain concentration was added and incubated for 2 hours. 1 g/mL Mega CD40L and 0.2 ng/mL LPS were added and incubated in an incubator at 37 C. with 5% CO.sub.2 for 20 hours. The cell supernatant was collected by centrifugation, and the TNF and IL-6 levels in the supernatant were detected using a kit. Dose-response data were fitted to an S-shaped curve using non-linear regression, and IC50 values were calculated.

    [0962] The results show that the ligand-drug conjugates of the present application could inhibit cytokine release from dendritic cells derived from human peripheral blood monocyte. The ligand-drug conjugates of the present application can inhibit inflammation, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    13.2 Evaluation of Drug Efficacy in LPS-Induced Acute Inflammation Model

    [0963] 8-10 week-old CD40 humanized female mice were randomly grouped and injected intraperitoneally with a concentration of the ligand-drug conjugates of the present application or the control ligand-drug conjugate. After 2-16 hours, the mice were injected intraperitoneally with LPS with a certain concentration. After 6 hours, 100 L of blood sample was collected, and the TNF, IL-6 and IL-10 contents were measured using a kit. After another 18 hours, the mice were euthanized, spleens were collected for single cell treatment, and the cells were stained with antibodies to determine the proportion of activated dendritic cells.

    [0964] The results show that the ligand-drug conjugates of the present application could inhibit the release of cytokines in an LPS-induced acute inflammation mouse model and reduce the proportion of activated dendritic cells in the spleen. The ligand-drug conjugates of the present application can inhibit inflammation, and can be used for preventing and/or treating diseases and/or conditions such as inflammation.

    Example 14: Pharmacokinetic and Toxicity Studies of ADCs at a Single Administration

    Objective

    [0965] To investigate the pharmacokinetic properties of the drug in monkeys and observe the toxic manifestation of the animals after a single intravenous drip of the ligand-drug conjugates (ADCs) of the present application in monkeys.

    Test Method

    [0966] Pharmacokinetics: after the single intravenous drip of the ligand-drug conjugates (ADCs) of the present application at difference doses in monkeys, blood samples were collected at a plurality of continuous time points, and the concentration of the drugs in the blood was detected by a specific detection method.

    [0967] Toxicity study: after the single intravenous drip of the ligand-drug conjugates (ADCs) of the present application at difference doses in monkeys, the tolerance of animals and drug-related toxicity toxic manifestation were investigated in multiple aspects such as clinical observation, body weight and food intake, hematology, blood biochemistry, urine and gross anatomy.

    [0968] The results show that: (1) after the ligand-drug conjugates (ADCs) of the present application were subjected to single intravenous drip on a monkey, the concentration of free drugs was very low, and the pharmacokinetic properties of a total antibody and the ligand-drug conjugates (ADCs) of the present application were similar, so that the ligand-drug conjugates (ADCs) of the present application were slowly released in the monkey, the coupling mode was stable, and the clinically planned administration frequency could be supported; and (2) after single intravenous drip of the ligand-drug conjugates (ADCs) of the present application in monkeys, animals had good tolerance and did not show serious or intolerable drug-related toxicity, which indicated that the ligand-drug conjugates (ADCs) of the present application were controllable in safety and could support further clinical application.

    [0969] The foregoing detailed description is provided by way of illustration and example, and is not intended to limit the scope of the appended claims. Various modifications of the embodiments currently enumerated in the present application will be apparent to those of ordinary skill in the art and are intended to be within the scope of the appended claims and their equivalents.