CXCR4 antagonists with amino acid skeleton, preparation therefor and biomedical use thereof

Abstract

The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt or prodrug thereof. ##STR00001##

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof: ##STR00056## wherein n.sub.1 and n.sub.2 are each 1 or 2, n.sub.3 is 3 or 4; R.sup.1 represents cyclohexyl; X represents —C(═O)NR.sup.2, wherein R.sup.2 represents hydrogen, or methyl; Y represents benzene or pyridine connected at para-positions; ##STR00057## A represents methyl, ##STR00058## A can form a monocyclic or bicyclic ring together with ; B represents hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, an amino acid substituent, or a group of formula (v) or (vi): ##STR00059## wherein B.sub.1 and B.sub.2 are each independently an optionally substituted cycloalkyl, aryl or heteroaryl; wherein (1) when B is the cycloalkyl, B is cyclopentyl, cyclohexyl, mono- or poly-substituted cyclohexyl methyl, pyridine-fused cyclohexyl; (2) when B is the aryl, B is phenyl, biphenyl, or naphthyl; (3) when B is the heteroaryl, B is imidazolyl, unsubstituted pyridyl, mono- or poly-substituted pyridyl; (4) when B is the amino acid substituent, B is an L- or D-α-amino acid; (5) when B is aryl-, heteroaryl-, saturated carbocyclic or heterocyclic ring-substituted alkyl or formyl, B is ##STR00060##

2. The compound according to claim 1, wherein ##STR00061## is an amino acid, and the amino acid is a natural or non-natural amino acid, wherein the amino acid is an α-amino acid, a β-amino acid, or a γ-amino acid, wherein a side chain of the α-amino acid is cyclized or not cyclized with an —NH.sub.2 terminal, and the amino acid comprises an L- or D-amino acid, wherein the α-amino acid comprises at least one selected from a group comprising arginine, lysine, citrulline, ornithine, histidine, aspartic acid, glutamic acid, glutamine, asparagine, serine, threonine, cysteine, phenylalanine, 3-(2-naphthyl) alanine, cyclohexylalanine, tryptophan, 5-hydroxytryptophan, tyrosine, glycine, phenylglycine, cyclohexylglycine, alanine, valine, leucine, isoleucine, methionine, proline, 4-hydroxyproline, 1-amino-1-cyclohexanecarboxylic acid and 1,2,3,4-tetrahydroisoquinolyl-3-carboxylic acid.

3. The compound according to claim 1, wherein A forms a monocyclic ring together with , and the compound of formula (I) has the following structures: ##STR00062##

4. The compound according to claim 1, wherein A forms a bicyclic ring together with , and the compound of formula (I) has the following structures: ##STR00063##

5. The compound according to claim 1, wherein the pharmaceutically acceptable salt comprises at least one selected from a group comprising trifluoroacetate, hydrochloride, acetate, sulfate, mesylate, tosylate, citrate, tartrate, fumarate, maleate and malate.

6. The compound according to claim 1, wherein B represents hydrogen, cyclohexyl, ##STR00064##

7. The compound according to claim 1, comprising one of the following structures: ##STR00065## ##STR00066## ##STR00067## ##STR00068## ##STR00069## ##STR00070## ##STR00071## or a pharmaceutically acceptable salt or prodrug thereof.

8. A pharmaceutical composition, comprising a compound according to claim 1 as an active ingredient and a pharmaceutically acceptable auxiliary.

9. The pharmaceutical composition according to claim 8, further comprising one or more other anti-HIV agents, wherein the other anti-HIV agent comprises at least one selected form a group comprising maraviroc, enfuvirtide, zidovudine, zalcitabine, stavudine, lamivudine, nevirapine, delavirdine, abacavir, efavirenz, tenofovir, emtricitabine, etravirine, and rilpivirine.

10. The pharmaceutical composition according to claim 9, further comprising cisplatin, cyclophosphamide, cytarabine, 5-fluorouracil, gemcitabine, taxol, docetaxel, adriamycin, glivec, tarceva, sorafenib, dasatinib, lapatinib, sunitinib, erlotinib, gefitinib, cetuximab, or herceptin of trastuzumab.

11. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is used for inhibiting interaction of a chemokine CXCL12 with a CXCR4 receptor or inhibiting the CXCR4 receptor.

12. The pharmaceutical composition according to claim 8, wherein inhibiting the CXCR4 receptor is realized by antagonizing the CXCR4 receptor, wherein: the pharmaceutical composition is used to block HIV from invading and infecting human target cells or to treat or prevent AIDS, or the pharmaceutical composition is used to mobilize human bone marrow hematopoietic cells, mesenchymal stem cells or stem cells, or the pharmaceutical composition is used to interfere with CXCL12/CXCR4-mediated cell migration and adhesion, or the pharmaceutical composition is used to prevent or treat CXCR4-mediated tumor metastasis, invasion or growth, or the pharmaceutical composition is used to block CXCL12/CXCR4-mediated autoimmune and inflammatory responses.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a graph showing experimental results of the inhibition of CXCR4 induced cell migration by compound 1 according to an embodiment of the present disclosure;

(2) FIG. 2 is a graph showing experimental results of an activity of compound 1 to inhibit intracellular calcium mobilization according to an embodiment of the present disclosure; and

(3) FIG. 3 is a graph showing experimental results of an activity of a positive agent AMD3100 to inhibit intracellular calcium mobilization according to an embodiment of the present disclosure.

DETAILED DESCRIPTION

(4) Examples of the present disclosure will be described in details below, which are exemplary and intended to explain the present disclosure, and shall not be construed to limit the present disclosure.

Example 1: Preparation of the Intermediate 1-3a: Refer to the Synthesis Scheme (1)

(5) 4-cyanobenzaldehyde (10 g) was dissolved in methanol (150 ml), to the obtained solution were added with N-(3-aminopropyl) cyclohexylamine (12.4 mL) and a suitable amount of 4 Å molecular sieves, and then the solution was heated to 65° C. and stirred for 6 hours. After cooled to the room temperature, the solution was added with sodium borohydride (5.8 g) in portions and stirred at room temperature for 1 to 2 hours. The reaction was quenched with a saturated ammonium chloride solution, and the solution was concentrated to dryness. The residue was ultrasonically washed with methanol and then filtered. The filtrate was collected and evaporated to dryness to obtain the intermediate 1-1a crude product for use in the next reaction directly.

(6) The above crude intermediate 1-1a was directly dissolved in tert-butanol (100 ml) and sodium carbonate solution (2 mol/L, 100 ml), and then added with Boc.sub.2O (40 g) in portions. The resulted solution was stirred overnight at room temperature, and then diluted with an appropriate amount of water, followed by extracted with ethyl acetate. The combined organic layer was washed with a saturated sodium chloride solution, and dried with anhydrous sodium sulfate as well as by rotary evaporation to obtain a residue. The residue was purified by silica gel column chromatography to obtain the intermediate 1-2a (22 g, yield: 63%).

(7) 7 g of the above intermediate 1-2a was weighed and dissolved in methanol (50 ml) and tetrahydrofuran (50 ml), followed by sequential addition of Raney Ni (wet weight, 2 g) and aqueous ammonia (2 mL) in nitrogen atmosphere, and then the nitrogen was replaced with hydrogen. The solution was stirred overnight in hydrogen atmosphere, and then filtered by diatomite pad. The filtrate was evaporated to dryness, and the residue obtained thereby is the intermediate 1-3a (6 g, yield: 85%), ESI-MS: [M+1].sup.+476.8.

Example 2: Preparation of the Intermediate 1-3b

(8) Referring to the synthesis scheme (1), the synthetic processes and conditions for the preparation of the intermediate 1-3b are similar to that of the intermediate 1-3a.

Example 3: Preparation of the Intermediate 2-3a: Refer to the Synthesis Scheme (2)

(9) 5 g of amino acid raw material, Fmoc-L-Lys(Boc)-OH, was weighted and suspended in dichloromethane (40 ml), and then added with trifluoroacetic acid (20 ml). The obtained solution was stirred at room temperature for 1 to 2 hours, and then dried by rotary evaporation. The residue was precipitated with ice diethyl ether to obtain a white precipitate (i.e., the intermediate 2-1a), which was directly dissolved in methanol (50 ml), and then added with acetone (20 ml) and trifluoroacetic acid (2 ml). The resulted solution, after stirred at room temperature for 2 hours, was added with sodium triacetoxyborohydride (11.3 g) and then stirred overnight. Finally, the solution was adjusted to a pH of 4 to 6 with a hydrochloric acid solution (4 mol/L), and then concentrated to a certain volume, followed by extracted with ethyl acetate. The combined organic phase was dried with anhydrous sodium sulfate and then filtered. The filtrate was dried by rotary evaporation to obtain a residue (i.e., a crude product of the intermediate 2-2a) for use in the next reaction directly.

(10) The above obtained intermediate 2-2a was dissolved in tert-butanol (50 mL) and sodium carbonate solution (2 mol/L, 20 mL), and then added with Boc.sub.2O (32 g) in portions. The solution was stirred overnight at room temperature, and then diluted with water, followed by extracted with ethyl acetate. The combined organic phase was washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and then filtered. The filtrate was dried by rotary evaporation, and the resulted residue was purified by silica gel column chromatography to obtain the intermediate 2-3a (3.35 g, yield: 61%), MALDI-TOF-MS [M+Na].sup.+: 533.2587

Example 4: Preparation of the Intermediate 2-3b

(11) Referring to the synthesis scheme (2), the synthetic processes and conditions for the preparation of the intermediate 2-3b are similar to that of the intermediate 2-3a. MALDI-TOF-MS: [M+Na].sup.+573.3250.

Example 5: Preparation of the Intermediate 2-3c

(12) Referring to the synthesis scheme (2), the synthetic processes and conditions for the preparation of the intermediate 2-3c are similar to that of the intermediate 2-3a. MALDI-TOF-MS: [M+Na].sup.+559.3325.

Example 6: Preparation of Compound 1: Refer to the Synthesis Scheme (3)

(13) Preparation of the intermediate 3-1: the intermediate 1-3a (3.20 g) and an amino acid raw material Fmoc-Arg(Pbf)-OH (4.58 g) were weighted and dissolved in DMF (50 ml), followed by addition of HATU (3.84 g), HOBt (0.91 g), and DIPEA (2.22 mL) successively. The resulted solution was stirred overnight at room temperature, and then added dropwise to water to form precipitates, which were collected and dried in vacuum to obtain a solid product (i.e., the intermediate 3-1) for use in the next reaction directly.

(14) Preparation of the intermediate 3-2: the above obtained intermediate 3-1 was directly dissolved in DMF (56 ml) and piperidine (14 ml), and stirred at room temperature for 1 to 2 hours. Then, the solution was diluted with a saturated sodium chloride solution, and extracted with ethyl acetate. The combined organic phase was washed with a saturated sodium chloride solution, dried with anhydrous sodium sulfate, and then filtered. The filtrate was dried by rotary evaporation, and the obtained residue was purified by silica gel column chromatography to obtain a semisolid, i.e., the intermediate 3-2 (4.3 g, yield: 72%), HR-MS: [M+Na].sup.+906.5131.

(15) Preparation of the intermediate 3-3: the above prepared intermediate 3-2 (200 mg) was weighted and dissolved in anhydrous methanol (5 ml), and then added with 2-pyridyl formaldehyde (0.023 mL) and acetic acid (0.010 ml). The resulted solution was stirred at room temperature for 2 hours, and then added with sodium triacetoxyborohydride (146 mg) to continue react overnight at room temperature. Finally, the solution was quenched with a saturated sodium carbonate solution and extracted with ethyl acetate. The combined organic phase was dried with anhydrous sodium sulfate, and then filtered. The filtrate was dried by rotary evaporation, and the obtained residue was purified by silica gel column chromatography to obtain an oily substance (i.e., the intermediate 3-3) for use in the next reaction directly.

(16) Preparation of target compound 1: the above prepared intermediate 3-3 was dissolved in a deprotection solution (trifluoroacetic acid:triisopropylsilane:water=95:2.5:2.5) to react for 2 hours at room temperature. The reaction solution, after removal of solvents therefrom, was precipitated and washed with ice diethyl ether. The collected solid was purified by reversed-phase medium pressure preparative chromatography to obtain trifluoroacetate of the compound 1 (45 mg).

(17) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.58; (br, 2H), 9.31; (s, 2H), 9.18; (t, J=5.5 Hz, 1H), 8.77; (br-s, 2H), 8.63; (d, J=4.7 Hz, 1H), 8.07; (d, J=4.5 Hz, 1H), 7.89; (t, J=7.7 Hz, 1H), 7.49-33; (m, 9H), 4.46; (dd, J=15.4, 6.0 Hz, 1H), 4.32; (dd, J=15.5, 5.2 Hz, 1H), 4.27; (br-s, 2H), 4.14; (s, 2H), 3.96; (t, J=6.0 Hz, 1H), 3.13; (dd, J=12.4, 6.2 Hz, 2H), 3.00; (br-s, 5H), 2.08-1.81; (m, 6H), 1.78-1.70; (m, 2H), 1.61-1.46; (m, 3H), 1.30-1.18; (m, 4H), 1.13-1.03; (m, 1H).

(18) .sup.1H NMR (400 MHz, DMSO-d.sub.6+D.sub.2O) δ 9.17; (t, J=5.7 Hz, 1H), 8.62; (d, J=4.5 Hz, 1H), 7.88; (td, J=7.7, 1.4 Hz, 1H), 7.48-7.42; (m, 4H), 7.33; (d, J=8.0 Hz, 2H), 4.44; (d, J=15.5 Hz, 1H), 4.31; (d, J=15.4 Hz, 1H), 4.26; (s, 2H), 4.13; (s, 2H), 3.12; (t, J=6.8 Hz, 2H), 3.06-2.89; (m, 5H), 2.05-1.82; (m, 6H), 1.77; (m, 2H), 1.64-1.42; (m, 3H), 1.32-1.15; (m, 4H), 1.15-1.02; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+523.3883.

Example 7: Preparation of Compound 2: Refer to the Synthesis Scheme (3)

(19) Preparation of the intermediate 3-4: the above prepared intermediate 3-2 (200 mg) and p-fluorobenzoic acid (39 mg) were weighted and dissolved in DMF (4 ml), followed by addition of HATU (131 g), HOBt (31 g), and DIPEA (0.076 mL) successively. The resulted solution was stirred overnight at room temperature, and then added dropwise to water to form precipitates, which were collected and dried in vacuum to obtain a solid product (i.e., the intermediate 3-4) for use in the next reaction directly.

(20) Preparation of target compound 2: the above prepared intermediate 3-4 was dissolved directly in a deprotection solution (trifluoroacetic acid:triisopropylsilane:water=95:2.5:2.5) to react for 2 hours at room temperature. The reaction solution, after removal of solvents therefrom, was precipitated and washed with ice diethyl ether. The collected solid was purified by reversed-phase medium pressure preparative chromatography to obtain trifluoroacetate of the compound 2 (145 mg).

(21) .sup.1H NMR (400 MHz, DMSO-d.sub.6) (δ 9.15 (br-s, 2H), 8.69; (br-s, 2H), 8.60; (dd, J=12.3, 6.8 Hz, 2H), 8.00; (dd, J=8.3, 5.8 Hz, 2H), 7.79; (br-s, 1H), 7.68-6.86; (m, 10H), 4.45; (dd, J=13.3, 8.7 Hz, 1H), 4.31; (qd, J=15.7, 6.0 Hz, 2H), 4.12; (br-s, 2H), 3.12; (dd, J=12.5, 6.4 Hz, 2H), 2.99; (br-s, 5H), 2.07-1.87; (m, 4H), 1.90-1.81; (m, 1H), 1.84-1.66; (m, 3H), 1.62-1.46; (m, 3H), 1.30-1.18; (m, 4H), 1.15-1.03; (m, 1H).

(22) .sup.1H NMR (400 MHz, DMSO-d.sub.6+D.sub.2O) δ 7.99-7.96; (m, 1H), 7.40; (d, J=8.2 Hz, 2H), 7.31-7.27; (m, 4H), 4.44; (dd, J=9.7, 5.0 Hz, 1H), 4.30; (q, J=15.7 Hz, 2H), 4.11; (s, 2H), 3.11; (t, J=7.0 Hz, 1H), 3.01-2.89; (m, 2H), 1.97-1.92; (m, 4H), 1.87-1.78; (m, 1H), 1.75-1.69; (m, 3H), 1.62-1.46; (m, 3H), 1.29-1.17; (m, 2H), 1.12-1.04; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+554.3613.

Example 8: Preparation of Compound 3

(23) Referring to the synthesis scheme (3), except that the intermediate 1-3a was replaced with benzylamine, the remaining synthetic processes and conditions for the preparation of compound 3 were similar to that of compound 1.

(24) .sup.1H NMR (400 MHz, D.sub.2O) (δ 8.64; (d, J=5.6 Hz, 1H), 8.42; (t, J=7.9 Hz, 1H), 8.00; (d, J=8.0 Hz, 1H), 7.97-7.84; (m, 1H), 7.25-7.13; (m, 5H), 4.58; (d, J=14.3 Hz, 1H), 4.43; (d, J=14.3 Hz, 1H), 4.30; (d, J=14.7 Hz, 1H), 4.12; (d, J=14.7 Hz, 1H), 4.03; (dd, J=7.6, 5.3 Hz, 1H), 2.94; (t, J=6.9 Hz, 2H), 1.88-1.77; (m, 2H), 1.40-1.28; (m, 2H). HRMS (ESI, m/z): [M+H].sup.+355.2254.

Example 9: Preparation of Compound 4

(25) Referring to the synthesis scheme (3), except that the intermediate 1-3a was replaced with the intermediate 1-3b, the remaining synthetic processes and conditions for the preparation of compound 4 were similar to that of compound 1.

(26) .sup.1H NMR (400 MHz, D.sub.2O) (δ 8.76; (d, J=5.5 Hz, 1H), 8.50; (t, J=7.8 Hz, 1H), 8.06; (d, J=8.0 Hz, 1H), 7.98; (t, J=6.7 Hz, 1H), 7.38; (d, J=7.8 Hz, 2H), 7.31; (d, J=7.9 Hz, 2H), 4.64; (d, J=14.3 Hz, 1H), 4.51; (d, J=14.3 Hz, 1H), 4.42; (d, J=15.0 Hz, 1H), 4.31; (d, J=15.0 Hz, 1H), 4.20-4.07; (m, 3H), 3.15-3.02; (m, 3H), 2.08-2.02; (m, 2H), 2.00-1.90; (m, 2H), 1.76; (d, J=12.4 Hz, 2H), 1.60-1.46; (m, 3H), 1.34-1.17; (m, 4H), 1.14-1.03; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+466.3289.

Example 10: Preparation of Compound 5

(27) The intermediate 3-2 (200 mg) mentioned in the synthesis scheme (3) was weighted and dissolved directly in a deprotection solution (trifluoroacetic acid:triisopropylsilane:water=95:2.5:2.5) to react for 2 hours at room temperature. The reaction solution, after removal of solvents therefrom, was precipitated and washed with ice diethyl ether. The collected solid was purified by reversed-phase medium pressure preparative chromatography to obtain trifluoroacetate of the compound 5 (145 mg).

(28) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.31; (br-s, 2H), 9.08; (t, J=5.6 Hz, 1H), 8.78; (br-s, 2H), 8.30; (br-s, 3H), 8.05; (s, 1H), 7.45; (d, J=8.1 Hz, 4H), 7.33; (d, J=8.0 Hz, 4H), 4.43; (dd, J=15.5, 6.1 Hz, 1H), 4.31; (dd, J=15.5, 5.4 Hz, 1H), 4.14; (br-s, 2H), 3.85; (d, J=5.0 Hz, 1H), 3.12; (dd, J=12.7, 6.5 Hz, 2H), 3.00; (br-s, 5H), 2.02-1.96; (m, 4H), 1.78-1.72; (m, 4H), 1.61-1.46; (m, 3H), 1.30-1.17; (m, 4H), 1.13-1.07; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+432.3431.

Example 11: Preparation of Compound 6

(29) Referring to the synthesis scheme (3), except that the Fmoc-Arg(Pbf)-OH was replaced with an amino acid raw material Fmoc-D-Arg(Pbf)-OH, the remaining synthetic processes and conditions for the preparation of compound 6 were similar to that of compound 1.

(30) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.73; (d, J=5.5 Hz, 1H), 8.47; (t, J=7.9 Hz, 1H), 8.03; (d, J=8.0 Hz, 1H), 7.99-7.92; (m, 1H), 7.36; (d, J=8.0 Hz, 2H), 7.30; (d, J=8.0 Hz, 2H), 4.55; (dd, J=51.9, 14.3 Hz, 2H), 4.35; (dd, J=38.7, 15.1 Hz, 2H), 4.15; (br-s, 2H), 4.13-4.07; (m, 1H), 3.10-2.95; (m, 7H), 2.03-1.89; (m, 6H), 1.72; (br-s, 2H), 1.56-1.43; (m, 3H), 1.26-1.43; (m, 4H), 1.10-1.04; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+523.3870.

Example 12: Preparation of Compound 7

(31) Referring to the synthesis scheme (3), except that p-fluorobenzoic acid was replaced with 4,4-difluorocyclohexanecarboxylic acid, the remaining synthetic processes and conditions for the preparation of compound 7 were similar to that of compound 2.

(32) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.12; (br-s, 2H), 8.65; (br-s, 2H), 8.50; (t, J=6.0 Hz, 1H), 8.12; (d, J=8.0 Hz, 1H), 7.76; (t, J=5.4 Hz, 1H), 7.41; (d, J=8.0 Hz, 2H), 7.28; (d, J=8.0 Hz, 2H), 7.30-6.95; (m, 4H), 4.36-4.31; (m, 1H), 4.27-4.22; (m, 2H), 4.13; (br-s, 2H), 3.11-3.06; (m, 2H), 2.99; (br-s, 5H), 2.41-2.35; (m, 1H), 2.08-1.92; (m, 6H), 1.85-1.66; (m, 7H), 1.64-1.43; (m, 6H), 1.29-1.18; (m, 4H), 1.14-1.06; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+578.3994.

Example 13: Preparation of Compound 8

(33) The synthetic processes and conditions for the preparation of compound 8 were similar to that of compound 1.

(34) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.45; (br., 2H), 9.28; (s, 2H), 9.21; (t, J=5.6 Hz, 1H), 8.74; (br-s, 2H), 8.05; (s, 1H), 7.66-7.20; (m, 11H), 4.44; (dd, J=15.3, 5.9 Hz, 1H), 4.32; (dd, J=15.3, 5.3 Hz, 1H), 4.14-4.02; (m, 4H), 3.79; (br-s, 1H), 3.11; (dd, J=12.6, 6.4 Hz, 2H), 2.99; (br-s, 5H), 1.97; (br-s, 4H), 1.92-1.67; (m, 4H), 1.66-1.35; (m, 3H), 1.28-1.17; (m, 4H), 1.15-1.05; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+540.3826.

Example 14: Preparation of Compound 9

(35) The synthetic processes and conditions for the preparation of compound 9 were similar to that of compound 1.

(36) .sup.1H NMR (400 MHz, D.sub.2O) δ 7.58; (q, J=8.5 Hz, 4H), 7.39; (d, J=7.7 Hz, 2H), 7.33; (d, J=7.8 Hz, 2H), 4.41-4.29; (m, 2H), 4.24-4.10; (m, 4H), 3.90; (t, J=6.5 Hz, 1H), 3.24-3.18; (m, 6H), 3.12-3.01; (m, 7H), 2.05-1.94; (m, 4H), 1.91-1.84; (m, 2H), 1.74; (br-s, 2H), 1.58-1.40; (m, 3H), 1.28-1.16; (m, 2H), 1.14-1.01; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+565.4337.

Example 15: Preparation of Compound 10

(37) The synthetic processes and conditions for the preparation of compound 10 were similar to that of compound 1.

(38) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.60; (br-s, 1H), 9.35; (br-s, 1H), 9.27; (br-s, 2H), 8.74; (br-s, 2H), 8.14; (d, J=8.1 Hz, 1H), 8.12-7.99; (m, 3H), 7.68-7.55; (m, 4H), 7.52-7.06; (m, 7H), 4.61-4.37; (m, 4H), 4.13; (br-s, 2H), 4.06; (br-s, 1H), 3.15-3.10; (m, 2H), 2.98; (br-s, 5H), 2.07-1.80; (m, 6H), 1.75; (br-s, 2H), 1.61-1.50; (m, 3H), 1.29-1.1.17; (m, 4H), 1.15-1.06; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+572.4095.

Example 16: Preparation of Compound 11

(39) The synthetic processes and conditions for the preparation of compound 11 were similar to that of compound 1.

(40) .sup.1H NMR (400 MHz, DMSO-d.sub.6) (δ 9.52; (br-s, 2H), 9.29-9.22; (m, 3H), 8.75; (br-s, 2H), 8.07; (t, J=5.7 Hz, 1H), 7.77-7.67; (m, 4H), 7.59-7.45; (m, 7H), 7.44-7.12; (m, 5H), 4.46; (dd, J=15.3, 6.0 Hz, 1H), 4.35; (dd, J=15.3, 5.4 Hz, 1H), 4.14; (br-s, 2H), 3.85; (br-s, 1H), 3.13; (dd, J=12.7, 6.5 Hz, 2H), 3.00; (br-s, 2H), 2.05-1.92; (m, 4H), 1.90-1.80; (m, 2H), 1.78-1.70; (m, 2H), 1.60; (d, J=12.2 Hz, 1H), 1.54-1.46; (m, 1H), 1.30-1.17; (m, 4H), 1.13-1.05; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+598.4236.

Example 17: Preparation of Compound 12

(41) The synthetic processes and conditions for the preparation of compound 12 were similar to that of compound 1.

(42) .sup.1H NMR (400 MHz, DMSO-d.sub.6) (δ 9.64; (br-s, 1H), 9.24; (br-s, 3H), 8.69; (br-s, 2H), 8.21; (br-s, 2H), 8.20; (s, 1H), 8.02; (s, 1H), 7.66-7.09; (m, 8H), 4.46-4.27; (m, 4H), 4.14; (br-s, 2H), 3.93; (br-s, 1H), 3.14; (dd, J=12.2, 6.1 Hz, 2H), 3.00; (br-s, 5H), 1.97-1.74; (m, 8H), 1.60; (d, J=12.1 Hz, 1H), 1.56-1.45; (m, 2H), 1.32-1.17; (m, 4H), 1.16-1.03; (m, 1H). HRMS (ESI, m/z): [M+H]+658.3674.

Example 18: Preparation of Compound 13

(43) The synthetic processes and conditions for the preparation of compound 13 were similar to that of compound 1.

(44) .sup.1H NMR (400 MHz, DMSO-d.sub.6) (δ 9.26; (br-s, 2H), 8.94; (s, 1H), 8.74; (br-s, 2H), 7.94; (d, J=5.3 Hz, 1H), 7.83-6.87; (m, 11H), 4.35; (d, J=3.7 Hz, 2H), 4.20; (br-s, 2H), 4.13; (br-s, 2H), 3.57; (br-s, 1H), 3.10; (d, J=6.1 Hz, 2H), 3.00; (br-s, 5H), 1.98; (br-s, 4H), 1.76-1.68; (m, 4H), 1.61-1.50; (m, 3H), 1.29-1.18; (m, 4H), 1.13-1.05; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+512.3829.

Example 19: Preparation of Compound 14

(45) The synthetic processes and conditions for the preparation of compound 14 were similar to that of compound 1.

(46) .sup.1H NMR (400 MHz, D.sub.2O) (δ 8.52; (d, J=5.3 Hz, 1H), 8.08; (d, J=7.8 Hz, 1H), 7.72-7.66; (m, 1H), 7.40; (d, J=8.0 Hz, 2H), 7.35; (d, J=8.0 Hz, 2H), 4.61; (t, J=4.9 Hz, 1H), 4.39; (s, 2H), 4.25; (dd, J=8.2, 5.1 Hz, 1H), 4.19; (s, 2H), 3.17-3.00; (m, 7H), 2.98-2.82; (m, 2H), 2.14-1.85; (m, 9H), 1.82-1.71; (m, 3H), 1.64-1.46; (m, 3H), 1.31-1.18; (m, 4H), 1.15-1.05; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+563.4183.

Example 20: Preparation of Compound 15

(47) The synthetic processes and conditions for the preparation of compound 15 were similar to that of compound 1.

(48) .sup.1H NMR (400 MHz, DMSO-d.sub.6) (δ 9.36; (t, J=5.8 Hz, 1H), 9.29; (br-s, 2H), 9.03-8.85; (m, 2H), 8.75; (br-s, 2H), 8.05; (t, J=5.6 Hz, 1H), 7.64-7.16; (m, 8H), 4.53; (dd, J=15.4, 6.4 Hz, 1H), 4.29; (dd, J=15.3, 5.0 Hz, 1H), 4.14; (br-s, 2H), 3.96; (br-s, 1H), 3.14; (dd, J=13.3, 7.0 Hz, 2H), 3.00; (br-s, 5H), 2.79; (br-s, 1H), 2.12-1.86; (m, 6H), 1.83-1.67; (m, 6H), 1.64-1.55; (m, 2H), 1.54-1.44; (m, 2H), 1.32-1.01; (m, 10H). HRMS (ESI, m/z): [M+H].sup.+514.4226.

Example 21: Preparation of Compound 16

(49) The synthetic processes and conditions for the preparation of compound 16 were similar to that of compound 1.

(50) .sup.1H NMR (400 MHz, D.sub.2O) (δ 7.43; (d, J=7.8 Hz, 1H), 7.38; (d, J=7.8 Hz, 1H), 4.44; (q, J=15.1 Hz, 2H), 4.21; (br-s, 2H), 3.84; (br-s, 1H), 3.20-2.97; (m, 7H), 2.87-2.82; (m, 1H), 2.68-2.63; (m, 1H), 2.09-2.00; (m, 4H), 1.94-1.86; (m, 2H), 1.79; (br-s, 2H), 1.70-1.40; (m, 9H), 1.35-1.05; (m, 8H), 0.99-0.82; (m, 2H). HRMS (ESI, m/z): [M+H].sup.+528.4393.

Example 22: Preparation of Compound 17

(51) Referring to the synthesis scheme (3), except that Fmoc-Lys(Boc)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 17 were similar to that of compound 1.

(52) .sup.1H NMR (400 MHz, D.sub.2O) (δ 8.62; (br-s, 1H), 8.08; (t, J=6.8 Hz, 1H), 7.63 (br-s, 2H), 7.45; (d, J=6.5 Hz, 2H), 7.38; (d, J=6.5 Hz, 2H), 4.50-4.30; (m, 4H), 4.24; (br-s, 2H), 3.99; (br-s, 1H), 3.25-3.00; (m, 5H), 2.99-2.85; (m, 2H), 2.15-1.89; (m, 6H), 1.88-1.74; (m, 2H), 1.73-1.56; (m, 3H), 1.47-1.21; (m, 6H), 1.20-1.08; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+495.3810.

Example 23: Preparation of Compound 18

(53) Referring to the synthesis scheme (3), except that the synthesized intermediate 2-3a was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 18 were similar to that of compound 1.

(54) .sup.1H NMR (400 MHz, D.sub.2O) (δ 8.76; (d, J=5.3 Hz, 1H), 8.47; (td, J=7.9, 1.1 Hz, 1H), 8.03; (d, J=8.0 Hz, 1H), 8.01-7.92; (m, 1H), 7.43; (d, J=8.1 Hz, 2H), 7.35; (d, J=8.1 Hz, 2H), 4.64; (d, J=14.3 Hz, 1H), 4.51; (d, J=14.3 Hz, 1H), 4.46-4.32; (m, 2H), 4.21; (br-s, 2H), 4.12; (t, J=6.4 Hz, 1H), 3.34-3.28; (m, 1H), 3.15-3.03; (m, 5H), 2.98-2.84; (m, 2H), 2.09-1.93; (m, 6H), 1.77; (br-s, 1H), 1.69-1.54; (m, 3H), 1.42-1.31; (m, 2H), 1.30-1.18; (m, 10H), 1.17-1.07; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+537.4280.

Example 24: Preparation of Compound 19

(55) Referring to the synthesis scheme (3), except that the synthesized intermediate 2-3b was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 19 were similar to that of compound 1.

(56) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.70; (d, J=5.0 Hz, 1H), 8.34; (t, J=7.8 Hz, 1H), 7.92-7.83; (m, 2H), 7.40; (d, J=8.0 Hz, 2H), 7.33; (d, J=8.0 Hz, 2H), 4.57-4.42; (m, 2H), 4.37; (s, 2H), 4.19; (s, 2H), 4.05; (t, J=6.3 Hz, 1H), 3.12-2.94; (m, 6H), 2.92-2.88; (m, 2H), 2.05-1.90; (m, 8H), 1.82-1.68; (m, 4H), 1.65-1.53; (m, 4H), 1.35-1.15; (m, 10H), 1.12-1.05; (m, 2H). HRMS (ESI, m/z): [M+H].sup.+577.4584.

Example 25: Preparation of Compound 20

(57) Referring to the synthesis scheme (3), except that the synthesized intermediate 2-3c was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 20 were similar to that of compound 1.

(58) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.59; (d, J=5.6 Hz, 1H), 8.36; (td, J=8.0, 1.3 Hz, 1H), 7.94; (d, J=8.0 Hz, 1H), 7.87-7.81; (m, 1H), 7.22; (d, J=8.1 Hz, 2H), 7.14; (d, J=8.1 Hz, 2H), 4.51 (d, J=14.3 Hz, 1H), 4.35; (d, J=14.3 Hz, 1H), 4.20; (s, 2H), 4.00; (s, 3H), 2.96-2.75; (m, 8H), 1.89-1.75; (m, 8H), 1.56-1.48; (m, 6H), 1.38; (d, J=12.2 Hz, 2H), 1.11-0.99; (m, 8H), 0.95-0.85; (m, 2H). HRMS (ESI, m/z): [M+H].sup.+563.4428.

Example 26: Preparation of Compound 21

(59) Referring to the synthesis scheme (3), except that Fmoc-His(Trt)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 21 were similar to that of compound 1.

(60) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.66; (d, J=5.6 Hz, 1H), 8.43; (t, J=7.9 Hz, 1H), 8.33; (s, 1H), 7.99; (d, J=8.0 Hz, 1H), 7.95-7.86; (m, 1H), 7.25; (d, J=7.9 Hz, 2H), 7.13-6.96; (m, 3H), 4.57; (d, J=14.5 Hz, 1H), 4.44; (d, J=14.5 Hz, 1H), 4.25; (d, J=14.8 Hz, 1H), 4.19; (dd, J=9.3, 5.5 Hz, 1H), 4.08; (s, 2H), 4.04; (d, J=14.9 Hz, 1H), 3.32; (dd, J=15.0, 5.3 Hz, 1H), 3.18; (dd, J=15.0, 9.4 Hz, 1H), 3.01-2.90; (m, 5H), 1.99-1.79; (m, 4H), 1.63; (br-s, 2H), 1.45; (d, J=12.3 Hz, 1H), 1.23-1.02; (m, 4H), 1.00-0.92; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+504.3451.

Example 27: Preparation of Compound 22

(61) Referring to the synthesis scheme (3), except that Fmoc-Asp(tBu)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 22 were similar to that of compound 1.

(62) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.49; (d, J=5.0 Hz, 1H), 8.05; (t, J=7.4 Hz, 1H), 7.60-7.54; (m, 2H), 7.30; (d, J=7.9 Hz, 2H), 7.21; (d, J=7.9 Hz, 2H), 4.35-4.18; (m, 4H), 4.10; (br-s, 2H), 4.01; (t, J=6.2 Hz, 1H), 3.05-2.93; (m, 5H), 2.85; (d, J=6.1 Hz, 2H), 1.96-1.84; (m, 4H), 1.68; (br-s, 2H), 1.50; (d, J=12.2 Hz, 1H), 1.21-1.09; (m, 4H), 1.05-0.95; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+482.3127.

Example 28: Preparation of Compound 23

(63) Referring to the synthesis scheme (3), except that Fmoc-Ala-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 23 were similar to that of compound 1.

(64) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.72; (d, J=5.7 Hz, 1H), 8.50; (t, J=7.9 Hz, 1H), 8.07; (d, J=8.0 Hz, 1H), 8.01-7.93; (m, 1H), 7.32; (d, J=8.0 Hz, 2H), 7.24; (d, J=8.0 Hz, 2H), 4.63 (d, J=14.4 Hz, 1H), 4.51; (d, J=14.3 Hz, 1H), 4.30; (dd, J=33.3, 15.4 Hz, 2H), 4.18; (q, J=7.0 Hz, 1H), 4.11; (s, 2H), 3.06-2.90; (m, 5H), 2.03-1.85; (m, 4H), 1.67; (d, J=6.0 Hz, 2H), 1.54-1.45; (m, 4H), 1.23-1.11; (m, 4H), 1.08-0.95; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+438.3235.

Example 29: Preparation of Compound 24

(65) Referring to the synthesis scheme (3), except that Fmoc-Ser(tBu)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 24 were similar to that of compound 1.

(66) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.53; (d, J=5.0 Hz, 1H), 8.04; (t, J=7.6 Hz, 1H), 7.63; (d, J=7.8 Hz, 1H), 7.61-7.52; (m, 1H), 7.35; (d, J=7.7 Hz, 2H), 7.26; (d, J=7.7 Hz, 2H), 4.43-4.25; (m, 4H), 4.15; (s, 2H), 4.08-3.95; (m, 2H), 3.92-3.85; (m, 1H), 3.11-2.90; (m, 5H), 2.05-1.86; (m, 4H), 1.72; (br-s, 2H), 1.54; (d, J=12.2 Hz, 1H), 1.25-1.13; (m, 4H), 1.09-1.03; (s, 1H). HRMS (ESI, m/z): [M+H].sup.+454.3179.

Example 30: Preparation of Compound 25

(67) Referring to the synthesis scheme (3), except that Fmoc-2-Nal-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 25 were similar to that of compound 1.

(68) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.66; (d, J=5.1 Hz, 1H), 8.18; (td, J=7.8, 1.3 Hz, 1H), 7.86; (d, J=7.8 Hz, 1H), 7.77-7.69; (m, 4H), 7.56-7.49; (m, 3H), 7.27 (d, J=8.4 Hz, 1H), 6.84; (d, J=8.0 Hz, 2H), 6.63; (d, J=8.0 Hz, 2H), 4.49; (q, J=14.4 Hz, 2H), 4.31-4.26; (m, 2H), 4.01; (q, J=13.1 Hz, 2H), 3.86; (d, J=15.0 Hz, 1H), 3.52; (dd, J=13.1, 5.2 Hz, 1H), 3.24-3.13; (m, 1H), 3.06-3.01; (m, 5H), 2.09-1.89; (m, 4H), 1.79; (br-s, 2H), 1.62; (d, J=12.0 Hz, 1H), 1.36-1.19; (m, 5H), 1.13-1.08; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+564.3712.

Example 31: Preparation of Compound 26

(69) Referring to the synthesis scheme (3), except that Fmoc-Trp(Boc)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 26 were similar to that of compound 1.

(70) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.73; (d, J=5.5 Hz, 1H), 8.43; (t, J=7.9 Hz, 1H), 7.98; (d, J=8.0 Hz, 1H), 7.95-7.89; (m, 1H), 7.48; (dd, J=7.8, 4.4 Hz, 2H), 7.26-7.20; (m, 3H), 7.14; (s, 1H), 7.08; (t, J=7.5 Hz, 1H), 6.82; (d, J=7.8 Hz, 2H), 4.61; (dd, J=34.7, 14.4 Hz, 2H), 4.33; (dd, J=9.5, 5.7 Hz, 1H), 4.26; (d, J=15.2 Hz, 1H), 4.19; (s, 2H), 4.02; (d, J=15.1 Hz, 1H), 3.50-3.45; (m, 1H), 3.40-3.34; (m, 1H), 3.14-3.00; (m, 5H), 2.08-1.98; (m, 4H), 1.79 (br-s, 2H), 1.62; (d, J=12.0 Hz, 1H), 1.32-1.21; (m, 4H), 1.17-1.10; (s, 1H). HRMS (ESI, m/z): [M+H].sup.+553.3651.

Example 32: Preparation of Compound 27

(71) Referring to the synthesis scheme (3), except that Fmoc-Tyr(tBu)-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 27 were similar to that of compound 1.

(72) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.50; (d, J=4.1 Hz, 1H), 7.92; (t, J=7.5 Hz, 1H), 7.47; (t, J=7.0 Hz, 2H), 7.22; (d, J=7.6 Hz, 2H), 6.86; (t, J=9.2 Hz, 4H), 6.58; (d, J=8.0 Hz, 2H), 4.32-4.23; (m, 3H), 4.12; (s, 2H), 3.98-3.87; (m, 2H), 3.18-3.13; (m, 1H), 3.08-2.92; (m, 5H), 2.90-2.80; (m, 1H), 2.05-1.82; (m, 4H), 1.69; (br-s, 2H), 1.52; (d, J=12.1 Hz, 1H), 1.25-1.10; (m, 4H), 1.08-0.99; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+530.3500.

Example 33: Preparation of Compound 28

(73) Referring to the synthesis scheme (3), except that Fmoc-Leu-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 28 were similar to that of compound 1.

(74) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.48; (d, J=4.7 Hz, 1H), 7.86; (t, J=7.5 Hz, 1H), 7.45-7.38; (m, 2H), 7.31; (d, J=7.9 Hz, 2H), 7.24; (d, J=7.9 Hz, 2H), 4.34; (d, J=15.1 Hz, 1H), 4.25-4.15; (m, 3H), 4.11; (s, 2H), 3.81; (dd, J=8.7, 5.8 Hz, 1H), 3.05-2.90; (m, 5H), 1.98-1.85; (m, 4H), 1.73-1.55; (m, 4H), 1.51; (d, J=12.1 Hz, 1H), 1.42-1.35; (m, 1H), 1.23-1.10; (m, 4H), 1.06-0.95; (m, 1H), 0.76; (d, J=6.4 Hz, 3H), 0.72; (d, J=6.5 Hz, 3H). HRMS (ESI, m/z): [M+H].sup.+480.3702.

Example 34: Preparation of Compound 29

(75) Referring to the synthesis scheme (3), except that Fmoc-Pro-OH was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 29 were similar to that of compound 1.

(76) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.45; (d, J=4.7 Hz, 1H), 7.95; (t, J=7.6 Hz, 1H), 7.64; (d, J=7.8 Hz, 1H), 7.56-7.46; (m, 1H), 7.39; (d, J=7.8 Hz, 2H), 7.19; (d, J=7.8 Hz, 2H), 4.67; (d, J=13.6 Hz, 1H), 4.46; (d, J=13.6 Hz, 1H), 4.40-4.29; (m, 1H), 4.21; (s, 2H), 4.19; (s, 2H), 3.85-3.76; (m, 1H), 3.43-3.36; (m, 1H), 3.17-3.00; (m, 5H), 2.62-2.49; (m, 1H), 2.27-2.13; (m, 1H), 2.09-1.92; (m, 6H), 1.77; (br-s, 2H), 1.59 (d, J=12.2 Hz, 1H), 1.32-1.17; (m, 4H), 1.15-1.05; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+464.3395.

Example 35: Preparation of Compound 30

(77) Referring to the synthesis scheme (3), except that Fmoc-L-1,2,3,4-tetrahydroxyisoquinoline-3-carboxylic acid was used as the amino acid raw material, the remaining synthetic processes and conditions for the preparation of compound 30 were similar to that of compound 1.

(78) .sup.1H NMR (400 MHz, D.sub.2O) δ 8.58; (d, J=5.2 Hz, 1H), 8.19; (td, J=7.9, 1.4 Hz, 1H), 7.75; (d, J=7.9 Hz, 1H), 7.73-7.65; (m, 1H), 7.41; (d, J=8.1 Hz, 1H), 7.35-7.20; (m, 5H), 7.18-7.10; (m, 2H), 4.64-4.34; (m, 3H), 4.33-4.12; (m, 5H), 3.37-3.30; (m, 1H), 3.26-3.16; (m, 1H), 3.15-3.05; (m, 5H), 2.12-1.91; (m, 4H), 1.78; (br-s, 2H), 1.61; (d, J=12.1 Hz, 1H), 1.39-1.17; (m, 4H), 1.19-1.01; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+526.3555.

Example 36: Preparation of Compound 31

(79) Referring to the synthesis scheme (3), the intermediate 1-3a (3.20 g) was condensed with a first amino acid raw material Fmoc-Arg(Pbf)-OH to obtain an intermediate, which was deprotected to remove the Fmoc protective group and then condensed with a second amino acid raw material Fmoc-Tyr(tBu)-OH, followed by removal of Fmoc, Pbf, tBu and Boc protective groups. The synthetic conditions were similar to that of compound 2.

(80) .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.46; (d, J=8.0 Hz, 2H), 7.38; (d, J=7.9 Hz, 2H), 7.24; (d, J=2.0 Hz, 1H), 7.10-7.01; (m, 1H), 6.85; (d, J=8.3 Hz, 1H), 6.72; (d, J=8.5 Hz, 1H), 4.53-4.30; (m, 3H), 4.20; (s, 2H), 4.15-4.55; (m, 1H), 3.26-3.00; (m, 8H), 2.98-2.90; (m, 1H), 2.18-2.03; (m, 4H), 1.95-1.80; (m, 3H), 1.80-1.56; (m, 4H), 1.41-1.28; (m, 4H), 1.25-1.17; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+595.4086.

Example 37: Preparation of Compound 32

(81) Referring to the synthesis scheme (3), the intermediate 1-3a (3.20 g) was condensed with a first amino acid raw material Fmoc-2-Nal-OH to obtain an intermediate, which was deprotected to remove the Fmoc protective group and then condensed with a second amino acid raw material Fmoc-Arg(Pbf)-OH, followed by removal of Fmoc, Pbf, tBu and Boc protective groups. The synthetic conditions were similar to that of compound 2.

(82) .sup.1H NMR (400 MHz, D.sub.2O) δ 7.87; (d, J=7.2 Hz, 1H), 7.79; (d, J=8.5 Hz, 1H), 7.77-7.71; (m, 1H), 7.59; (s, 1H), 7.56-7.47; (m, 2H), 7.36; (d, J=8.5 Hz, 1H), 6.86; (d, J=8.1 Hz, 2H), 6.69; (d, J=8.0 Hz, 2H), 4.68; (dd, J=10.3, 6.3 Hz, 1H), 4.33; (d, J=15.3 Hz, 1H), 4.08-3.97; (m, 3H), 3.94; (d, J=15.3 Hz, 1H), 3.31; (dd, J=13.3, 6.2 Hz, 1H), 3.15-2.98; (m, 8H), 2.06-1.93; (m, 4H), 1.92-1.84; (m, 2H), 1.82-1.72; (m, 2H), 1.66-1.49; (m, 3H), 1.33-1.18; (m, 4H), 1.17-1.04; (m, 1H). HRMS (ESI, m/z): [M+H].sup.+629.4282.

Example 38 Cytotoxicity Assay of Compounds

(83) 1. Experimental method: the experiment was carried out using HEK293 cell line. The HEK293 cells cultured in DMEM medium (containing 10% fetal bovine serum, 100 IU penicillin and 0.1 mg/ml streptomycin) were trypsinized, counted, and plated in a 96-well plate with each well containing 100 μl suspension and including 3×10.sup.3 cells in total. After culturation in a constant-temperature incubator containing 5% CO.sub.2 at 37° C. overnight, different concentrations of the compound were added. After 72 hours of incubation in total, 20 μl CellTiter 96 was added to further incubate for 1 to 4 hours, and then an absorption value at 490 nm was measured on a microplate reader. The changes in cell viability prior to and subsequent to the addition of the compound were observed by comparing the values of the blank control group and the experimental group.
2. Experimental results: as shown in Table 1, the compounds have no cytotoxic effect and do not have inhibitory activity on cytogenesis at a concentration of 50 μM.

Example 39 Binding Affinity Assay of Compounds on CXCR4 Receptor Protein

(84) 3. Experimental method: the cell line used for the affinity activity assay of the CXCR4 was a CHO cell line constructed in vitro and stably transfected with CXCR4. The CHO cell line cultured in DMEM medium (containing 10% fetal bovine serum, 100 IU penicillin, 0.1 mg/ml streptomycin and 0.2 mg/ml G418) was trypsinized, washed, counted, and then suspended in an FACS buffer (PBS containing 0.5% BSA and 0.05% NaN.sub.3), followed by being plated in a 96-well plate. Each cell contains 100 μl of the reaction system, which includes 5×10.sup.5 cells, 250 ng/ml 12G5 monoclonal antibody and different concentrations of compound for detection. After incubated on ice for 40 min and washed with the FACS buffer, the reaction system was added with the secondary antibody (anti-mouse FITC-labeled IgG) for further incubation on ice for 30 min and then washed twice with the FACS buffer. Absorption values at 485 nmEX/535 nmEM were measured on a microplate reader (PerkinElmer), and fluorescence intensities were recorded. For each compound, a percentage of the compound that inhibits the binding of the 12G5 monoclonal antibody to the receptor CXCR4, i.e. a binding inhibition ratio at 0.1 μM, was calculated.
4. Experimental results: as shown in Table 1, most of the compounds show a strong inhibitory activity at the concentration of 0.1 μM. The inhibition ratio of the compounds is divided into the following intervals: 75% to 100% for A, 50% to 74% for B, 25% to 49% for C, and 0 to 24% for D.

(85) TABLE-US-00001 TABLE 1 Inhibition ratio interval of Cytotoxicity Compound compounds assay No. Chemical structural formula at 0.1 μM (IC.sub.50, μM) 1 A >50 2 B >50 3 D NT 4 D >50 5 C >50 6 B >50 7 0 B >50 8 B >50 9 A >50 10 B >50 11 B >50 12 D >50 13 B >50 14 D NT 15 B >20 16 A >50 17 0 A >50 18 A >50 19 A >50 20 B >50 21 A >50 22 B >50 23 A >50 24 A >50 25 B >10 26 A >50 27 0 B >50 28 C >50 29 A >50 30 A >20 31 A >50 32 A >50 Note: “−” indicates no inhibitory activity, and “NT” indicates without detection.

Example 40 Assay on the Inhibition of Compounds on CXCR4 Induced Cell Migration

(86) 1. Experimental method: the assay on the CXCR4 induced cell migration was detected using SupT1 cell line which naturally expresses the CXCR4. SupT1 cells cultured and suspended in RPMI 1640 (containing 10% fetal bovine serum, 100 IU penicillin and 0.1 mg/ml streptomycin) were collected and then washed with RPMI 1640 containing 0.5% BSA, after which the cells are counted and suspended to a concentration of 3.0×10.sup.7 cells/ml, and then uniformly mixed with different concentrations of compound. Then, 75 μl of the obtained mixture was taken and added to a tranwell insert, and incubated at 37° C. for 30 min. Subsequently, 200 μl of RPMI 1640 containing 0.5% BSA of 2 nM SDF-1α was added to each well of a lower culture plate, and a group to be used as a background value did not contain SDF-1α. After incubation at 37° C. for 3 hours, the tranwell insert was removed, and 40 μl of CellTiter 96 (Promega) was added to each cell of the lower culture plate. After 1 to 4 hours of incubation, absorption values at 490 nm were detected. The experimental results of each group were obtained by at least three independent experiments, and the inhibition curve was plotted by the GraphPad software.
2. Experimental results: as shown in FIG. 1, in the CXCR4 induced cell migration inhibiting experiments, the exemplified compound 1 has a significant inhibiting ability to the cell migration, which exhibits a good concentration-dependent relationship, indicating that the compounds according to embodiments of the present disclosure have a significant inhibiting ability to the cell migration, and such inhibiting ability is concentration-dependent.

Example 41 Assay on the Inhibition of Compounds on Intracellular Calcium Mobilization

(87) 3. Experimental method: the cell line used in this assay was SupT1. Prior to the experiment, the cells were collected and washed with an experimental buffer (an HBSS buffer containing 20 mM HEPES), and then incubated with a buffer containing 4 μM Fluo-4 and 1 mM probenecid sodium for 30 min at 37° C., after which the residual dye was washed away, and the cells were suspended to a concentration of 2×10.sup.6 cells/ml. Afterwards, 200 μl of the cell suspension was taken and added to a black 96-well plate, and absorption values of the cells at 494 nmEX/516 nmEM were recorded. Subsequently, different concentrations of compound were added and mixed uniformly, and the baseline was recorded. Thereafter, 50 nM of the SDF-1α was added and mixed uniformly. Signals of calcium ions were recorded, and the differences in fluorescence intensity between a group without addition of the compound and groups added with different concentrations of the compound were observed.
4. Experimental results: as shown in FIG. 2, the exemplified compound 1 can completely inhibit the generation of signal of the calcium mobilization at a concentration of 0.2 μM, while the positive agent AMD3100 fails to inhibit the signal of the calcium mobilization at a concentration of 1.0 μM (as shown in FIG. 3). As can be seen from the results of the exemplified compound 1, the CXCR4 antagonistic activity exhibited by the compound according to embodiments of the present disclosure is superior to that of AMD3100.

(88) Reference throughout this specification to “an embodiment,” “some embodiments,” “one embodiment”, “an example,” “a specific example,” or “some examples,” means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present disclosure. Thus, the appearances of the phrases such as “in an embodiment,” “in some embodiment”, “in one embodiment”, “in an example,” “in a specific example,” or “in some examples,” in various places throughout this specification are not necessarily referring to the same embodiment or example of the present disclosure. Furthermore, the particular features, structures, materials, or characteristics may be combined in any suitable manner in one or more embodiments or examples. In addition, in the absence of contradiction, different embodiments or examples of the present disclosure or features described in different embodiments or examples may be combined by those skilled in the art.

(89) Although embodiments of the present disclosure have been shown and described above, it would be appreciated by those skilled in the art that the above embodiments are exemplary, cannot be construed to limit the present disclosure, and changes, modifications, alternatives and variants can be made in the embodiments without departing from the scope of the present disclosure.