Imidazoquinoline amine derivatives, pharmaceutical compositions and therapeutic methods thereof

Abstract

The invention provides novel imidazoquinoline amine derivatives, having agonistic activities to Toll-like receptors (TLRs), in particular TLR7 and/or TLR8, pharmaceutical compositions thereof, and methods of treatment, reduction or prevention of certain diseases or conditions mediated by or associated with TLR7 and/or TLR8, e.g., graft rejection, autoimmunity, inflammation allergy, asthma, infection, sepsis, cancer and immunodeficiency.

Claims

1. A compound having the structural formula (I): ##STR00061## wherein, R.sup.1 is a C.sub.1-C.sub.8 alkyl group; R.sup.2 is (CH.sub.2).sub.m, wherein m is an integer selected from 1 to 8; L is a linking moiety comprising a group selected from: ##STR00062## and R.sup.3 is a C.sub.1-C.sub.32 alkyl group; or a pharmaceutically acceptable form or an isotope derivative thereof.

2. The compound of claim 1, wherein R.sup.1 is a C.sub.3-C.sub.6 alkyl group, m is an integer selected from 3 to 8, and R.sup.3 is a C.sub.12-C.sub.32 alkyl group.

3. The compound of claim 2, wherein R.sup.1 is a C.sub.4 alkyl group, having the structural formula: ##STR00063##

4. The compound of claim 3, wherein m is 4, having the structural formula: ##STR00064##

5. The compound of claim 1, wherein L comprises a group selected from: ##STR00065##

6. The compound of claim 1, having the structural formula of: ##STR00066##

7. The compound of claim 1, having the structural formula of: ##STR00067##

8. The compound of claim 1, having the structural formula of: ##STR00068##

9. The compound of claim 1, having the structural formula of: ##STR00069##

10. The compound of claim 1, having the structural formula of: ##STR00070##

11. The compound of claim 1, having the structural formula of: ##STR00071##

12. The compound of claim 1, having the structural formula of: ##STR00072##

13. The compound of claim 1, having the structural formula of: ##STR00073##

14. The compound of claim 1, having the structural formula of: ##STR00074##

15. The compound of claim 1, having the structural formula of: ##STR00075##

16. The compound of claim 1, having the structural formula of: ##STR00076##

17. The compound of claim 1, having the structural formula of: ##STR00077##

18. A pharmaceutical composition comprising a compound of claim 1.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) The invention provides novel imidazoquinoline amine derivatives, which are agonists of TLRs, in particular TLR7 and/or TLR8, pharmaceutical compositions thereof, and methods of treatment, reduction or prevention of certain diseases or conditions mediated by or associated with TLR7 and/or TLR8, e.g., graft rejection, autoimmunity, inflammation allergy, asthma, infection, sepsis, cancer and immunodeficiency, or related diseases and conditions.

(2) In one aspect, the invention generally relates to a compound having the structural formula (I):

(3) ##STR00004##
wherein,

(4) R.sup.1 is a C.sub.1-C.sub.8 (e.g., C.sub.1-C.sub.4, C.sub.4-C.sub.6, C.sub.6-C.sub.8) alkyl group;

(5) R.sup.2 is (CH.sub.2).sub.m, wherein m is an integer selected from 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8);

(6) L is a linking moiety; and

(7) R.sup.3 is a C.sub.1-C.sub.32 (e.g., C.sub.1-C.sub.16, C.sub.16-C.sub.32, C.sub.16-C.sub.24) alkyl group;

(8) or a pharmaceutically acceptable form or an isotope derivative thereof.

(9) In certain embodiments, R.sup.1 is a C.sub.3-C.sub.6 (e.g., C.sub.3, C.sub.4, C.sub.5, C.sub.6) alkyl group, m is an integer selected from 3 to 8 (e.g., 3, 4, 5, 6, 7, 8), and R.sup.3 is a C.sub.12-C.sub.32 (e.g., C.sub.1-C.sub.16, C.sub.16-C.sub.32, C.sub.16-C.sub.24) alkyl group.

(10) In certain embodiments, R.sup.1 is a C.sub.4 alkyl group, having the structural formula:

(11) ##STR00005##

(12) In certain embodiments, R.sup.1 is a C.sub.4 alkyl group and m is 4, having the structural formula:

(13) ##STR00006##

(14) In certain embodiments, R.sup.2 is a C.sub.14-C.sub.20 (e.g., C.sub.14-C.sub.16, C.sub.16-C.sub.18, C.sub.18-C.sub.20) alkyl group.

(15) In certain embodiments, L comprises an amino group.

(16) In certain embodiments, L is an acyclic group.

(17) In certain embodiments, L comprises a 3- to 7-(e.g., 3-, 4-, 5-, 6-, 7-) membered ring.

(18) In certain embodiments, L is selected from:

(19) ##STR00007##

(20) It is noted that the exemplary linkers (L) may be connected to the rest of the compound such that L is boned to R.sup.2 on the left and to R.sup.3 on the right, or may be connected to the rest of the compound such that L is bonded to R.sup.2 on the right and to R.sup.3 on the left.

(21) Exemplary compounds of the invention include, but are not limited to, those listed in Table 1, or a pharmaceutically acceptable form or an isotope derivative thereof.

(22) TABLE-US-00001 TABLE 1 Exemplary Compounds 1 2 0 3 4 5 6 7 8 9 10 11 12

(23) In certain embodiments, the compound is

(24) ##STR00020##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(25) In certain embodiments, the compound is

(26) ##STR00021##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(27) In certain embodiments, the compound is

(28) ##STR00022##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(29) In certain embodiments, the compound is

(30) ##STR00023##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(31) In certain embodiments, the compound is

(32) ##STR00024##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(33) In certain embodiments, the compound is

(34) ##STR00025##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(35) In certain embodiments, the compound is

(36) ##STR00026##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(37) In certain embodiments, the compound is

(38) ##STR00027##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(39) In certain embodiments, the compound is

(40) ##STR00028##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(41) In certain embodiments, the compound is

(42) ##STR00029##
or a pharmaceutically acceptable form or an isotope derivative thereof.

(43) In certain embodiments, the compound is

(44) ##STR00030##

(45) In certain embodiments, the compound is

(46) ##STR00031##

(47) In another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein.

(48) In yet another aspect, the invention generally relates to a pharmaceutical composition comprising an amount of a compound having the structural formula (I):

(49) ##STR00032##
wherein,

(50) R.sup.1 is a C.sub.1-C.sub.8 (e.g., C.sub.1-C.sub.4, C.sub.4-C.sub.6, C.sub.6-C.sub.8) alkyl group;

(51) R.sup.2 is (CH.sub.2).sub.m, wherein m is an integer selected from 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8);

(52) L is a linking moiety; and

(53) R.sup.3 is a C.sub.1-C.sub.32 (e.g., C.sub.1-C.sub.16, C.sub.16-C.sub.32, C.sub.16-C.sub.24) alkyl group;

(54) or a pharmaceutically acceptable form or an isotope derivative thereof, effective to treat, prevent, or reduce one or more diseases or disorders, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent.

(55) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent an autoimmune disease, or a related disease or disorder.

(56) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent graft rejection, or a related disease or disorder.

(57) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent an allergy, or a related disease or disorder.

(58) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent an immunodeficiency, or a related disease or disorder.

(59) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent infection, sepsis, or a related disease or disorder.

(60) In certain embodiments, the pharmaceutical composition disclosed herein is effective to treat, reduce or prevent cancer, or a related disease or disorder.

(61) In yet another aspect, the invention generally relates to a unit dosage form comprising a compound disclosed herein.

(62) In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition disclosed herein.

(63) In yet another aspect, the invention generally relates to a method for treating, reducing, or preventing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound having the structural formula (I):

(64) ##STR00033##
wherein,

(65) R.sup.1 is a C.sub.1-C.sub.8 alkyl group;

(66) R.sup.2 is (CH.sub.2).sub.m, wherein m is an integer selected from 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8);

(67) L is a linking moiety; and

(68) R.sup.3 is a C.sub.1-C.sub.32 (e.g., C.sub.1-C.sub.16, C.sub.16-C.sub.32, C.sub.16-C.sub.24) alkyl group;

(69) or a pharmaceutically acceptable form or an isotope derivative thereof, effective to treat, prevent, or reduce one or more of autoimmune diseases, graft rejection, allergies, immunodeficiency, infection, sepsis, cancer, or a related disease or disorder thereof, in a mammal, including a human, and a pharmaceutically acceptable excipient, carrier, or diluent.

(70) In certain embodiments, the method is to treat, reduce or prevent an autoimmune disease, or a related disease or disorder.

(71) In certain embodiments, the method is to treat, reduce or prevent graft rejection, or a related disease or disorder.

(72) In certain embodiments, the method is effective to treat, reduce or prevent allergy, or a related disease or disorder.

(73) In certain embodiments, the method is effective to treat, reduce or prevent an immunodeficiency, or a related disease or disorder.

(74) In certain embodiments, the method is effective to treat, reduce or prevent infection and/or sepsis, or a related disease or disorder

(75) In certain embodiments, the method is effective to treat, reduce or prevent cancer, or a related disease or disorder.

(76) In yet another aspect, the invention generally relates to a method for modulating immune response, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

(77) In yet another aspect, the invention generally relates to a method for modulating TLR7- and/or TLR8-mediated signaling, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

(78) In yet another aspect, the invention generally relates to a method for of treating or reducing a condition or disorder treatable by modulation of TLR7- and/or TLR8-mediated cellular activities, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein.

(79) In yet another aspect, the invention generally relates to a method for treating, reducing, or preventing a disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein, wherein the disease or disorder is autoimmune diseases, graft rejection, allergies, immunodeficiency, infection, sepsis, cancer, or a related disease or disorder.

(80) In certain embodiments, the compound is an agonist of TLR7.

(81) In certain embodiments, the compound is an agonist of TLR8.

(82) In yet another aspect, the invention generally relates to use of a compound disclosed herein for treating, reducing or preventing a disease or disorder.

(83) In yet another aspect, the invention generally relates to use of a compound disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating, reducing or preventing a disease or disorder.

(84) In certain embodiments, the disease or disorder is mediated by or associated with TLR7- and/or TLR8-mediated signaling.

(85) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent a disease or disorder selected from autoimmune diseases, graft rejection, allergies, immunodeficiency, infection, sepsis, cancer, or a related disease or disorder thereof.

(86) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent an autoimmune disease, or a related disease or disorder.

(87) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent graft rejection, or a related disease or disorder.

(88) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent allergy, or a related disease or disorder.

(89) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent immunodeficiency, or a related disease or disorder.

(90) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent infection and/or sepsis, or a related disease or disorder

(91) In certain embodiments, the compound disclosed herein is used to treat, reduce or prevent cancer, or a related disease or disorder.

(92) In certain embodiments, the compound disclosed herein is used as vaccine adjuvants and to prevent, inhibit or treat a variety of disorders like inflammation, cancer and microbial infections.

(93) The compounds according to the invention may be used as agonists of TLR's, specifically for TLR7 and TLR8.

(94) The compounds according to the invention may provide methods for modulating TLR7- and/or TLR8-mediated signaling. The methods of the invention are useful, for example, when it is desirable to alter TLR7- and/or TLR8-mediated signaling in response to a suitable TLR7 and/or TLR8 ligand or a TLR7 and/or TLR8 signaling agonist.

(95) The compounds according to the invention may be used in the treatment or prevention of conditions and disorders include, but are not limited to, cancer, immune complex-associated diseases, inflammatory disorders, immunodeficiency, graft rejection, graft-versus-host disease, allergies, asthma, infection, and sepsis. More specifically, methods useful in the treatment of conditions involving autoimmunity, inflammation, allergy, asthma, graft rejection. Alternatively, methods useful in the treatment of conditions involving infection, cancer, and immunodeficiency generally will employ compounds disclosed herein that augment TLR7- and/or TLR8-mediated signaling in response to a suitable TLR7 and/or TLR8 ligand. In some instances, the compositions can be used to inhibit or promote TLR 7- and/or TLR8-mediated signaling in response to a TLR7 and/or TLR8 ligand or signaling agonist. In other instance, the compositions can be used to inhibit or promote TLR7- and/or TLR8-mediated immune-stimulation in a subject.

(96) The compounds according to the inventions may also be used in the treatment or prevention of hepatocarcinomas, cholangiocarcinoma and malignant mesothelioma, pancreatic cancer, head and neck cancer, and haemoangioma.

(97) The compounds according to the inventions may also be used in the treatment or prevention treating obesity in a patient. The invention provides a method of treating type II diabetes in a patient in need of treatment comprising administering to the patient an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof. Preferably the patient is a human. The present invention provides a method for treating nonalcoholic steatohepatitis in a patient in need of such treatment, comprising administering to the patient an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof.

(98) The present invention provides use of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of obesity. The present invention provides the use of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treatment to provide therapeutic weight loss.

(99) The amount of compound in compositions of this invention is such that is effective to measurably modulate TLR's, in particular TLR7 and/or TLR8, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, the amount of compound in compositions of this invention is such that is effective to measurably modulate TLR's, or a mutant thereof, in a biological sample or in a patient. In certain embodiments, a composition of this invention is formulated for administration to a patient in need of such composition.

(100) Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compounds described herein or derivatives thereof are admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (i) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid, (ii) binders, as for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (iii) humectants, as for example, glycerol, (iv) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (v) solution retarders, as for example, paraffin, (vi) absorption accelerators, as for example, quaternary ammonium compounds, (vii) wetting agents, as for example, cetyl alcohol, and glycerol monostearate, (viii) adsorbents, as for example, kaolin and bentonite, and (ix) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethyleneglycols, and the like. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others known in the art.

(101) Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifiers, such as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid esters of sorbitan, or mixtures of these substances, and the like. Besides such inert diluents, the composition can also include additional agents, such as wetting, emulsifying, suspending, sweetening, flavoring, or perfuming agents.

(102) Materials, compositions, and components disclosed herein can be used for, can be used in conjunction with, can be used in preparation for, or are products of the disclosed methods and compositions. It is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutations of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a method is disclosed and discussed and a number of modifications that can be made to a number of molecules including in the method are discussed, each and every combination and permutation of the method, and the modifications that are possible are specifically contemplated unless specifically indicated to the contrary. Likewise, any subset or combination of these is also specifically contemplated and disclosed. This concept applies to all aspects of this disclosure including, but not limited to, steps in methods using the disclosed compositions. Thus, if there are a variety of additional steps that can be performed, it is understood that each of these additional steps can be performed with any specific method steps or combination of method steps of the disclosed methods, and that each such combination or subset of combinations is specifically contemplated and should be considered disclosed.

(103) Compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, intratumoral and intracranial injection or infusion techniques.

(104) Pharmaceutically acceptable compositions of this invention are orally administered in any orally acceptable dosage form. Exemplary oral dosage forms are capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents are optionally also added.

(105) Alternatively, pharmaceutically acceptable compositions of this invention are administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

(106) Pharmaceutically acceptable compositions of this invention are also administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

(107) Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches are also used.

(108) For topical applications, provided pharmaceutically acceptable compositions are formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Exemplary carriers for topical administration of compounds of this aremineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

(109) Pharmaceutically acceptable compositions of this invention are optionally administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and are prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

(110) Most preferably, pharmaceutically acceptable compositions of this invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.

(111) The amount of compounds of the present invention that are optionally combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of the compound can be administered to a patient receiving these compositions.

(112) It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the present invention in the composition will also depend upon the particular compound in the composition.

(113) The following examples are meant to be illustrative of the practice of the invention and not limiting in any way.

EXAMPLES

(114) The below Examples describe certain exemplary embodiments of compounds prepared according to the disclosed invention. It will be appreciated that the following general methods, and other methods known to one of ordinary skill in the art, can be applied to compounds and subclasses and species thereof, as disclosed herein.

(115) .sup.1H NMR was recorded on a Bruker 400 MHz spectrometer, using residual signal of deuterated solvent as internal reference. Chemical shifts () are reported in ppm relative to the residual solvent signal (=2.50 ppm for 1H NMR in DMSO-d6; =3.31 ppm for 1H NMR in MeOD-d4).

(116) LCMS was taken on a quadrupole Mass Spectrometer on Shimadzu LC/MS2020 Series (SunFire C18 3.5 m 50*4.6 mm) operating in ES (+) or () ionization mode; T=40 C.; flow rate=2.0 mL/min; detected wavelength: 254 nm.

(117) HPLC was performed under conditions: (Flash: Welchrom C18 5 um 4.6150 mm); Wavelength 254 nm and 214 nm; Mobile phase: A water (0.03% TFA); B MeCN (0.03% TFA); Flow rate: 1 mL/min; Injection volume: 2 L; Run time: 16 min; Equilibration: 6 min.

(118) Prep-HPLC was performed under conditions: (Flash: Welchrom C18 25019 mm); Wavelength 254 nm and 214 nm; Mobile phase: A water (0.1% HCl); B MeCN; Flow rate: 20 mL/min; Injection volume: 0.5 mL; Run time: 18 min; Equilibration: 2 min.

Example 1. N1-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-N2-heptadecyloxalamide (1)

(119) ##STR00034##

(120) ##STR00035## ##STR00036##

(121) Step 1: To a stirred solution of 1-1 (22.0 g, 247 mmol) in DCM (300 mL) under argon atmosphere were added pyridine (29 mL, 371 mmol) and benzyl chloroformate (42.3 g, 247 mmol, in 250 mL DCM) at 0 C.; the mixture was warmed to RT and stirred for 16 h. The reaction mixture was diluted with 1N HCl (400 mL) at 0 C. The organic layer was washed with brine (400 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford 1-2 (29 g, yield: 52.7%) as a light-yellow solid. .sup.1H NMR (400 MHz, CDCl3) 7.38-7.31 (m, 5H), 5.09 (s, 2H), 3.75-3.55 (m, 2H), 3.29-3.16 (m, 2H), 1.58-1.55 (m, 4H).

(122) Step 2: A solution of N-hydroxyphthalimide (23.3 g, 143 mmol), benzyl (4-hydroxybutyl)carbamate 1-2 (29.0 g, 130 mmol) and triphenylphosphine (51.0 g, 195 mmol) in dichloromethane (600 mL) was chilled in an ice bath and a solution of diisopropylazodicarboxylate (DIAD, 39.4 g, 195 mmol) in dichloromethane (250 mL) was slowly added with stirring. The reaction mixture was stirred at 0 C. for 1 h and warm to RT, stirred for 4 h. The reaction was completed by TLC(PE/EA=2:1). The reaction was concentrated under reduced pressure, the residue was purified by column chromatography (PE/EA=2:1) to give 1-3 (32 g, yield: 68.2%) as a white solid. .sup.1H NMR (400 MHz, CDCl3) 7.84-7.81 (m, 2H), 7.77-7.72 (m, 2H), 7.38-7.28 (m, 5H), 5.09 (s, 2H), 4.23 (t, J=5.2 Hz, 2H), 3.34-3.29 (m, 2H), 1.84-1.75 (m, 4H).

(123) Step 3: To a solution of 1-3 (22.0 g, 59.8 mmol) in EtOH (350 mL) was added hydrazine (6.2 g, in 11.5 mL water) and the reaction was stirred at RT overnight. The reaction was cooled in the freezer and the resulting solid was removed by filtration. The solid was washed with ethanol (400 mL). The combined filtrate was concentrated under reduced pressure and diethyl ether (400 mL) was added. Insoluble impurities were removed by filtration and 3.0 M HCl in dioxane (100 mL) was added to the solution. A precipitate formed immediately. The mixture was stirred at RT for 16 h, the solid was recovered by filtration and dried under vacuum to give 1-4 (12 g, yield: 85.7%) as a white solid. .sup.1H NMR (400 MHz, DMSO) 7.36-7.27 (m, 5H), 5.35 (s, 2H), 5.09 (s, 2H), 4.91 (s, 1H), 3.66 (t, J=6.2 Hz, 2H), 3.24-3.19 (m, 2H), 1.62-1.55 (m, 4H). MS m/z (ESI): 239.1 [M+H].sup.+.

(124) Step 4: To a solution of 1-4 (10.0 g, 38.2 mmol) in isopropanol (20 mL) was added 1-4a (11.8 g, 49.6 mmol) and TEA (7.72 g, 76.4 mmol), the reaction mixture was stirred at 100 C. for 16 h under argon atmosphere. The mixture was concentrated under vacuum, the residue was purified by column chromatography (DCM/MeOH=10:1) to give 1-5 (14.5 g, yield: 85.3%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 9.25 (s, 1H), 8.76 (d, J=4.2 Hz, 1H), 8.49-8.37 (m, 1H), 7.87 (d, J=2.0 Hz, 2H), 7.38-7.31 (m, 5H), 5.12 (s, 2H), 4.58-4.35 (m, 2H), 3.44-3.35 (m, 2H), 3.11-3.00 (m, 2H), 2.18-2.06 (m, 2H), 2.02-1.94 (m, 2H), 1.92-1.82 (m, 2H), 1.03 (t, J=6.4 Hz, 3H).

(125) Step 5: To a solution of 1-5 (13.5 g, 30.3 mmol) in DCM (300 mL) was added 30% H.sub.2O.sub.2 (67.5 mL) and m-CPBA (12.3 g, 60.6 mmol) at 0 C., the reaction mixture was allowed to stirred at RT for 16 h under argon atmosphere. LCMS (MC18-197-066-1) showed 55% starting material remained. m-CPBA (6.2 g, 15.2 mmol) was added to the mixture, then stirred at r.t. for 2 h; LCMS showed 38% starting material remained. m-CPBA (12.3 g, 60.6 mmol) was added to the mixture, then stirred at RT for 16 h; LCMS showed 17% starting material remained. Water (200 mL) added to the mixture, and extracted with DCM (200 mL*2), the combined organic layer was washed with sodium carbonate solution (400 mL) and brine (400 mL), dried over sodium sulfate, filtered and concentrated in vacuum to give 1-6 (13 g, crude) which was used in next step directly. MS m/z (ESI): 463.2 [M+H].sup.+.

(126) Step 6: To a solution of 1-6 (13 g, 28.1 mmol) in DCM (300 mL) was added ammonium hydroxide (13 mL of a28-30 percent aqueous solution) at 0 C., a solution of TsCl (13.4 g, 70.3 mmol) dissolved in 50 mL dichloromethane was slowly added with vigorous stirring. The cooling bath was removed and the reaction was stirred for an additional 16 h. The desired product was detected by LCMS. The mixture was added water (300 mL) and extracted with DCM (200 mL*2), the combined organic layer was washed with sodium carbonate solution (300 mL*2) and brine (400 mL*2), dried over sodium sulfate, filtered and concentrated in vacuo, the residue was purified by column chromatography (DCM/MeOH=10:1) to afford 1-7 (7.0 g, yield: 54.3%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl3) 8.11 (d, J=8.0 Hz, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.65-7.61 (m, 1H), 7.49-7.45 (m, 1H), 7.40-7.36 (m, 5H), 5.14 (s, 2H), 4.96 (s, 1H), 4.36 (t, J=6.0 Hz, 2H), 3.41-3.36 (m, 2H), 2.96 (t, J=7.6 Hz, 2H), 2.05-2.00 (m, 2H), 1.93-1.83 (m, 4H), 1.53-1.47 (m, 2H), 1.02 (t, J=7.2 Hz, 3H). MS m/z (ESI): 462.6 [M+H]+.

(127) Step 7: A solution of 1-7 (7.0 g, 15.2 mmol) in con.HCl (20 mL) was stirred at 80 C. overnight. LCMS showed the reaction was completed. The mixture was cooled to RT, added water (30 mL), adjusted pH to 9-10 with sodium carbonate solid, extracted with DCM/MeOH=10:1 (50 mL*6). The combined organic layer was dried by sodium sulfate, filtered, and concentrated in vacuo to give 1-8 (5.0 g, crude) as a yellow oil which was used in next step directly. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.10 (d, J=8.0 Hz, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.34 (t, J=7.6 Hz, 1H), 5.36 (s, 2H), 4.32 (t, J=6.8 Hz, 2H), 2.98-2.92 (m, 2H), 2.86 (t, J=6.8 Hz, 2H), 2.02 (dd, J=15.2, 6.8 Hz, 2H), 1.91 (dd, J=15.2, 7.4 Hz, 2H), 1.78-1.73 (m, 2H), 1.71-1.62 (m, 2H), 1.52-1.46 (m, 2H), 1.00 (t, J=7.2 Hz, 3H). MS m/z (ESI): 328.2 [M+H].sup.+.

(128) Step 8: To a solution of 1-9a (0.51 mL, 5.88 mmol) in dry DCM (20 mL) was added 1-9 (1.0 g, 3.92 mmol) in dry DCM (20 mL) dropwise at 0 C., then the mixture was stirred at 0 C. for 1 h, DIEA (507 mg, 3.92 mmol) was added dropwise, the mixture was stirred at room temperature for 4 h. The mixture was concentrated in vacuo to give the desired product 1-10 (1.0 g, crude), which was used in next step directly.

(129) Step 9: To a solution of 1-10 (1.0 g, crude) in dry DCM (10 mL) was added 1-8 (100 mg, 0.306 mmol) in dry DCM (10 mL) dropwise at 0 C., DIEA (79 mg, 0.612 mmol) was added dropwise, the mixture was stirred at RT for 16 h. he mixture was filtered, the filtrate was concentrated in vacuo, the residue was purified by prep-TLC (DCM/MeOH=10:1) to give compound 1 (39.18 mg, yield: 20.1%) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d6) 8.87 (t, J=6.0 Hz, 1H), 8.70 (t, J=6.0 Hz, 1H), 8.13 (d, J=8.0 Hz, 1H), 7.97 (s, 2H), 7.74 (d, J=8.4 Hz, 1H), 7.63 (t, J=7.6 Hz, 1H), 7.48 (t, J=7.6 Hz, 1H), 4.38 (t, J=6.4 Hz, 2H), 3.28-3.23 (m, 2H), 3.12 (dd, J=13.4, 6.8 Hz, 2H), 2.98 (t, J=7.6 Hz, 2H), 1.95-1.87 (m, 2H), 1.84 (dd, J=15.0, 7.6 Hz, 2H), 1.76 (dd, J=15.0, 7.6 Hz, 2H), 1.43 (dd, J=14.9, 7.4 Hz, 4H), 1.22 (br.s, 28H), 0.94 (t, J=7.6 Hz, 3H), 0.84 (d, J=13.6 Hz, 3H). MS m/z (ESI) 637.4 [M+H].sup.+.

Example 2. 3-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)amino)-4-(heptadecylamino)cyclobut-3-ene-1,2-dione (2)

(130) ##STR00037##

(131) ##STR00038##

(132) Step 1: To a solution of 2-1 (200 mg, 1.177 mmol) in MeOH (6 mL) was added 2-2 (300 mg, 1177 mmol) in MeOH (10 mL) at 0 C. dropwise, then the mixture was stirred at RT for 4 h. TLC (PE/EA=2:1) showed the reaction was completed. The mixture was concentrated in vacuo to give the desired product 2-3 (520 mg, yield: 91.4%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.82-4.77 (m, 2H), 3.43-3.41 (m, 2H), 1.64-1.59 (m, 3H), 1.46 (t, J=6.8 Hz, 2H), 1.32-1.26 (m, 28H), 0.88 (t, J=6.8 Hz, 3H).

(133) Step 2: To a solution of 2-3 (198 mg, 0.413 mmol) and 1-8 (90 mg, 0.275 mmol) in MeOH (10 mL) was added DIEA (71 mg, 0.55 mmol), the mixture was stirred at RT for 16 h. The mixture was concentrated in vacuo, the residue was purified by prep-HPLC (FA) to give the desired product 2 (FA salt), which was neutralized with sodium carbonate solution (10 mL) to give the product 2 as free base (20.8 mg, yield: 10.3%) as a light yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3, DMSO-d6) 7.92 (d, J=6.4 Hz, 1H), 7.60 (d, J=6.8 Hz, 1H), 7.36 (s, 1H), 7.19 (d, J=6.4 Hz, 1H), 6.91 (s, 1H), 6.79 (s, 1H), 5.48 (s, 2H), 4.21 (d, J=16.0 Hz, 2H), 3.62 (d, J=2.0 Hz, 2H), 3.48 (d, J=1.9 Hz, 2H), 2.82-2.76 (m, 2H), 1.94-1.86 (m, 2H), 1.80-1.69 (m, 4H), 1.47-1.40 (m, 2H), 1.34 (d, J=6.4 Hz, 2H), 1.10 (br.s, 26H), 0.87-0.83 (m, 2H), 0.76-0.67 (m, 6H). MS m/z (ESI): 661.4 [M+H].sup.+.

Example 3. N-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-1-hexadecylpiperidine-4-carboxamide (3)

(134) ##STR00039##

(135) ##STR00040##

(136) Step 1: To a solution of NaH (510 mg, 12.74 mmol) in dry DMF (10 mL) was added 3-1 (1.0 g, 6.37 mmol) at 25 C. The reaction mixture was stirred for 30 min at 25 C. followed by addition of 3-1a (2.91 g, 9.55 mmol). The mixture was allowed to warm to RT and stirred overnight. After diluting with H.sub.2O (100 mL), the mixture was extracted with EA (ethyl acetate, 100 mL3). The combined organics were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (PE/EA=100/1-10/1) to give 3-2 (1.5 g, yield: 45.3%) as light yellow oil, .sup.1H NMR (400 MHz, CDCl3) 4.15-4.10 (m, 2H), 2.89 (d, J=5.5 Hz, 2H), 2.32-2.27 (m, 3H), 1.99-1.89 (m, 4H), 1.79-1.78 (m, 2H), 1.48 (s, 2H), 1.30-1.23 (m, 29H), 0.88 (t, J=5.5 Hz, 3H).

(137) Step 2: To a solution of 3-2 (500 mg, 6.37 mmol) in EtOH (25 mL) was added NaOH/H.sub.2O (4N,15 mL). The reaction solution was stirred at RT overnight. The mixture was concentrated to remove most of ethanol. The aqueous phase was neutralized with HCl (4N, 15 mL). The crude product stirred with EtOH (25 mL) for 1 h, the solid was collected by filtration and dried to give 3-3 (350 mg, yield: 75.6%) as white solid. .sup.1H NMR (400 MHz, MeOD) 3.45-3.43 (m, 2H), 3.04-2.97 (m, 4H), 2.38 (s, 1H), 2.10-2.07 (m, 2H), 1.92 (s, 2H), 1.87 (s, 2H), 1.31-1.26 (m, 26H), 0.90 (t, J=5.5 Hz, 3H).

(138) Step 3: To a solution of 3-3 (233 mg, 0.612 mmol) in dry DCM (25 mL) was add 1-8 (100 mg, 0.306 mmol), 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC, 116 mg, 0.612 mmol) and DMAP (38 mg, 0.306 mmol). The mixture was stirred at RT for 2 h. After diluting with H.sub.2O (100 mL), the mixture was extracted with DCM (100 mL*3). The organic phase was washed with brine, dried with Na.sub.2SO.sub.4 and concentrated. The residue was purified by Pre-HPLC to give Compound 3 (16.12 mg, yield: 8%) as off-white solid. .sup.1H NMR (400 MHz, DMSO-d6) 8.03 (d, J=5.5 Hz, 1H), 7.80 (s, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.47-7.43 (m, 1H), 7.29-7.25 (m, 1H), 6.60 (s, 2H), 4.32 (s, 2H), 3.15 (d, J=8.2 Hz, 2H), 2.96-2.92 (m, 2H), 2.83 (d, J=7.2, 7.2 Hz, 2H), 2.22-2.18 (m, 2H), 2.05 (s, 1H), 1.88-1.78 (m, 6H), 1.66-1.54 (m, 6H), 1.46-1.23 (m, 30H), 0.95 (t, J=5.5 Hz, 3H), 0.85 (t, J=5.5 Hz, 3H). MS m/z (ESI): 663.5 [M+H].sup.+.

(139) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.02 (s, 1H), 7.80 (s, 1H), 7.75 (q, J=17.0 Hz, 7.5 Hz, 2H), 7.35 (d, J=7.5 Hz, 2H), 7.09 (s, 1H), 7.06 (m, 2H), 4.17 (m, 2H), 4.02 (t, J=1.0 Hz, 1H), 3.73 (q, 11.0 Hz, 3 Hz, 1H), 3.22 (m, 2H), 2.82 (m, 2H), 2.75 (t, J=3.0 Hz, 1H), 2.57 (m, 2H), 2.39 (m, 1H), 2.27 (s, 3H). 2.09 (m, 2H), m/z: 478.2 [M+H].sup.+.

Example 4. N-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-1-hexadecylpiperidine-4-carboxamide (4)

(140) ##STR00041##

(141) ##STR00042## ##STR00043## ##STR00044##

(142) Step 1: To a solution of 4-1 (30.0 g, 184 mmol) and 4-1a (70.1 g, 552 mmol) in CH.sub.3CN (500 mL) were added TEA (37.2 g, 368 mmol) and KI (3.05 g, 18.4 mmol), the mixture was stirred at 70 C. for 24 h. After concentration, the residue was purified by column chromatography (PE/EA=8:1) to afford 4-2 (32 g, yield: 70%) as a white solid. .sup.1H NMR (400 MHz, CDCl3) 7.85-7.82 (M, 2H), 7.79-7.73 (m, 2H), 4.25 (t, J=6.0 Hz, 2H), 3.68 (t, J=6.4 Hz, 2H), 2.11-2.03 (m, 2H), 1.99-1.92 (m, 2H).

(143) Step 2: To a solution of 4-2 (60 g, 238 mmol) in EtOH (1500 mL) was added hydrazine (16.8 g, in 30 mL water) and the reaction was stirred at RT overnight. The reaction was cooled in the freezer and the resulting solid was removed by filtration. The solid was washed with ethanol (400 mL). The combined filtrate was concentrated under reduced pressure. The residue was mixed with diethyl ether (500 mL). Insoluble impurities were removed by filtration and 4.0 M HCl in 1,4-dioxane (100 mL) was added to the filtrate. A precipitate formed immediately. The mixture was stirred at RT for 2 h, the solid was recovered by filtration and dried under vacuum to give 4-3 (18.2 g, yield: 62.3%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) 11.06 (s, 2H), 4.03 (t, J=6.0 Hz, 2H), 3.67 (t, J=6.4 Hz, 2H), 1.80-1.69 (m, 4H).

(144) Step 3: To a solution of 4-4 (20 g, 112 mmol) in pyridine (50 mL) was added Pyr.HCl (1.34 g) and 4-4a (41.6 g, 224 mmol) in 60 mL pyridine. The mixture was stirred at RT overnight. After concentration, the residue was diluted with 10% sodium carbonate solution (100 mL) and stirred for 30 min. The solid was collected and triturated with PE/EA=5:1 (100 mL) to give 4-5 (25 g, yield: 86.2%) as a yellow solid. .sup.1H NMR (400 MHz, DMSO-d6) 10.07 (s, 1H), 9.04 (s, 1H), 8.20 (dd, J=8.4, 1.2 Hz, 1H), 8.09 (dd, J=8.4, 0.8 Hz, 1H), 7.86-7.75 (m, 2H), 2.54-2.42 (m, 2H), 1.71-1.49 (m, 2H), 1.48-1.35 (m, 2H), 0.94 (t, J=7.2 Hz, 3H).

(145) Step 4: A solution of 4-5 (2.5 g, 9.54 mmol) and 4-3 (2.5 g, 15.5 mmol) in isopropanal (12 mL) was stirred at 85 C. for 5 h under microwave. After concentration, the residue was purified by column chromatography (EA) to afford 4-6 (1.75 g, yield: 56%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 9.29 (s, 1H), 9.05 (d, J=7.0 Hz, 1H), 8.50 (d, J=6.8 Hz, 1H), 8.15-7.79 (m, 2H), 4.60-4.43 (m, 2H), 3.72 (t, J=5.0 Hz, 2H), 3.10 (t, J=7.2 Hz, 2H), 2.32-2.26 (m, 2H), 2.20-2.12 (m, 2H), 2.02-1.97 (m, 2H), 1.58-1.52 (m, 2H), 1.04 (t, J=7.2 Hz, 3H).

(146) Step 5: To a solution of 4-6 (11 g, 33.2 mmol) in DCM (400 mL) was added 30% H.sub.2O.sub.2 (55 mL) and m-CPBA (20.2 g, 60.6 mmol) at 0 C., the reaction mixture was allowed to stirred at RT for 16 h under argon atmosphere. Water (200 mL) added to the mixture, and extracted with DCM (200 mL*2). The combined organic layer was washed with sodium carbonate solution (400 mL) and brine (400 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give 4-7 (10 g, crude), which was used in next step directly. MS m/z (ESI): 348.1 [M+H].sup.+.

(147) Step 6: To a solution of 4-7 (5.0 g, 14.4 mmol) in DCM (100 mL) was added aqueous ammonium hydroxide (5 mL) at 0 C., a solution of TsCl (6.84 g, 36.0 mmol) in 40 mL dichloromethane was slowly added with vigorous stirring. The cooling bath was removed and the reaction was stirred for an additional 16 h. The mixture was diluted with sodium carbonate solution (100 mL) and extracted with DCM (200 mL*2). The combined organic layer was washed with brine (400 mL*2), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (EA/MeOH=10:1) to afford 4-8 (3.2 g, yield: 64%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.09 (d, J=8.0 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.45 (t, J=7.6 Hz, 1H), 4.37 (t, J=6.4 Hz, 2H), 3.69 (t, J=6.0 Hz, 2H), 2.98-2.92 (m, 2H), 2.14-2.11 (m, 2H), 1.93-1.88 (m, 4H), 1.52-1.47 (m, 2H), 1.01 (t, J=7.2 Hz, 3H). MS m/z (ESI): 347.2 [M+H].sup.+.

(148) Step 7: A solution of 4-8 (1.0 g, 2.89 mmol) in 4-8a (2 mL) was stirred at 70 C. for 6 h. LCMS showed 60% product. The mixture was diluted with water (10 mL) and extracted with EA (50 mL*10). The combined organic layer was washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford 4-9 (500 mg) as a yellow oil which was used in next step directly. MS m/z (ESI): 413.2 [M+H].sup.+.

(149) Step 8: To a solution of 4-9 (230 mg, 0.558 mmol) in DCM (10 mL) were added stearic acid (332 mg, 1.116 mmol), EDC (212 mg, 1.116 mmol) and DMAP (68 mg, 0.558 mmol), the mixture was stirred at RT overnight. The mixture was diluted with DCM (20 mL), washed with saturated ammonium chloride (40 mL) and brine (30 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (FA) to give the desired product (FA salt), which was neutralized with sodium carbonate solution (10 mL) and further purified by prep-TLC (DCM/MeOH=10:1) to give the free base 4 (26 mg, yield: 5.6%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.12 (t, J=8.4 Hz, 1H), 7.93 (d, J=8.4 Hz, 1H), 7.63 (dd, J=8.4, 7.2 Hz, 1H), 7.50 (t, J=7.6 Hz, 1H), 4.44-4.33 (m, 2H), 3.53-3.44 (m, 3H), 2.99-2.89 (m, 3H), 2.73 (s, 3H), 2.51-2.48 (m, 1H), 2.36-2.31 (m, 3H), 2.34 (dd, J=14.2, 6.6 Hz, 2H), 2.17-2.11 (m, 2H), 2.04-1.97 (m, 4H), 1.92-1.80 (m, 4H), 1.66-1.61 (m, 2H), 1.53-1.47 (m, 2H), 1.38-1.16 (m, 26H), 1.03-0.93 (m, 3H), 0.88 (t, J=6.8 Hz, 3H) MS m/z (ESI): 679.5 [M+H].sup.+.

Example 5. N1-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-N3,N3-dimethyl-N1-octadecylpropane-1,3-diamine (5)

(150) ##STR00045##

(151) ##STR00046##

(152) A solution of 4-9 (90 mg, 0.218 mmol) and 5-1 (67 mg, 0.262 mmol) in DCM (12 mL) was stirred at RT for 1 h, NaBH(OAc).sub.3 (138 mg, 0.654 mmol) was added to the mixture and stirred for another 3 h. The mixture was concentrated in vacuo and the residue was purified by prep-TLC (DCM/MeOH=10:1) to give the title product 5 (54.4 mg, yield: 29.3%) as a light-yellow oil. .sup.1H NMR (400 MHz, MeOD) 8.27 (dd, J=8.0, 0.8 Hz, 1H), 7.76 (dd, J=8.4, 0.8 Hz, 1H), 7.72-7.66 (m, 1H), 7.58-7.53 (m, 1H), 4.49 (t, J=6.5 Hz, 2H), 3.06 (dd, J=10.3, 4.8 Hz, 2H), 3.03 (d, J=7.4 Hz, 2H), 3.00-2.97 (m, 2H), 2.89 (dd, J=9.9, 6.0 Hz, 2H), 2.75 (s, 6H), 2.10-2.05 (m, 2H), 1.99-1.91 (m, 6H), 1.71-1.62 (m, 2H), 1.53 (dd, J=15.1, 7.5 Hz, 2H), 1.39-1.37 (m, 4H), 1.33-1.23 (m, 28H), 1.03 (t, J=7.2 Hz, 3H), 0.89 (t, J=6.8 Hz, 3H). MS m/z (ESI): 665.5 [M+H].sup.+.

Example 6. N1-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-N1-(3-(dimethylamino)propyl)-N2-heptadecyloxalamide (6)

(153) ##STR00047##

(154) ##STR00048##

(155) Step 1: To a solution of 4-9 (150 mg, 0.364 mmol) in dry DCM (15 mL) was added 6-1 (53.3 mg, 0.437 mmol) in dry DCM (2 mL) dropwise at 0 C., DIEA (141 mg, 1.092 mmol) was added dropwise. The mixture was stirred at RT for 16 h. The mixture was diluted with H.sub.2O (10 mL) and extract with DCM (20 mL*3). The combined organic phase was washed with brine, dried with Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep-TLC (DCM/MeOH=6:1) to give the desired product 6-2 (80 mg, yield: 44.2%) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.08 (d, J=7.6 Hz, 1H), 7.92-7.87 (m, 1H), 7.60 (t, J=7.6 Hz, 1H), 7.47 (t, J=7.6 Hz, 1H), 4.36 (t, J=6.0 Hz, 2H), 3.89 (d, J=5.2 Hz, 2H), 3.55 (t, J=6.8 Hz, 2H), 3.44 (dd, J=13.8, 6.5 Hz, 2H), 3.00-2.93 (m, 2H), 2.78-2.69 (m, 2H), 2.57 (s, 3H), 2.52-2.48 (m, 2H), 2.39 (s, 6H), 2.01 (d, J=8.8 Hz, 2H), 1.91 (dd, J=8.0, 2.8 Hz, 2H), 1.57-1.51 (m, 2H), 1.01 (t, J=7.2 Hz, 3H). MS m/z (ESI): 499.2 [M+H].sup.+.

(156) Step 2; To a solution of 6-2 (60 mg, 0.12 mmol) in MeOH/H.sub.2O=4:1 (5 mL) was added NaOH (24 mg, 0.60 mmol). The reaction solution was stirred at RT for 30 min. The mixture was concentrated in vacuo, the residue was diluted with water and adjusted to pH=5-6 with HCl. The aqueous was extracted with DCM/MeOH=10:1(30 mL2), dried and concentrated in vacuo to give 6-3 (50 mg, crude) as white solid, which was used in next step directly. MS m/z (ESI): 485.6 [M+H].sup.+.

(157) Step 3; To a solution of 6-3 (50 mg, 0.103 mmol) in DCM/DMF=4:1 (5 mL) were added 6-3a (52.9 mg, 0.206 mmol), EDCI (39.1 mg, 0.206 mmol) and HOBt (27.8 mg, 0.206 mmol). The mixture was stirred at room temperature overnight. The mixture was concentrated in vacuo, the residue was purified by prep-HPLC (HCl) to give the compound 6 (10.03 mg, yield: 13.5%, HCl salt) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 14.84 (s, 1H), 12.42 (s, 1H), 9.01 (s, 1H), 8.13 (d, J=1.6 Hz, 1H), 7.93 (s, 1H), 7.63 (s, 1H), 7.52 (d, J=8.0 Hz, 1H), 6.89 (s, 1H), 4.50-4.39 (m, 2H), 4.02-3.84 (m, 2H), 3.67-3.51 (m, 2H), 3.30-3.10 (m, 4H), 3.04-2.94 (m, 2H), 2.88-2.86 (m, 4H), 2.39-2.29 (m, 2H), 2.09-1.98 (m, 4H), 1.91-1.89 (m, 2H), 1.56-1.51 (m, 4H), 1.30-1.25 (m, 30H), 1.01 (t, J=6.0 Hz, 3H), 0.88 (t, J=6.8 Hz, 3H). MS m/z (ESI): 722.6 [M+H].sup.+.

Example 7. 3-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)(3-(dimethylamino)propyl)amino)-4-(heptadecylamino)cyclobut-3-ene-1,2-dione (7)

(158) ##STR00049##

(159) ##STR00050##

(160) To a solution of 4-9 (230 mg, 0.558 mmol) and 7-1 (400 mg, 0.837 mmol) in MeOH (10 mL) was added DIEA (144 mg, 1.116 mmol). The mixture was stirred at RT for 16 h. The mixture was concentrated in vacuo, the residue was purified by prep-TLC (DCM/MeOH=10:1) to give the desired product 7 (21.03 mg, yield: 4.9%) as a light-yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.61 (s, 1H), 8.03 (d, J=8.0 Hz, 1H), 7.80 (d, J=8.0 Hz, 1H), 7.50 (t, J=7.6 Hz, 1H), 7.41 (t, J=7.6 Hz, 1H), 4.37 (t, J=6.4 Hz, 2H), 3.82 (d, J=6.4 Hz, 2H), 3.72-3.65 (m, 2H), 3.60-3.47 (m, 2H), 2.91 (t, J=7.6 Hz, 2H), 2.75-2.65 (m, 2H), 2.44 (s, 6H), 2.05-2.01 (m, 2H), 1.93-1.91 (m, 2H), 1.87-1.81 (m, 2H), 1.65-1.58 (m, 2H), 1.45 (dd, J=15.0, 7.5 Hz, 2H), 1.27-1.22 (m, 30H), 0.96 (t, J=7.2 Hz, 3H), 0.84 (d, J=7.0 Hz, 3H). MS m/z (ESI): 746.5 [M+H].sup.+.

Example 8. 2-butyl-1-(4-(4-heptadecyl-1H-1,2,3-triazol-1-yl)butoxy)-1H-imidazo[4,5-c]quinolin-4-amine (8)

(161) ##STR00051##

(162) ##STR00052##

(163) Step 1: To a solution of 8-1 (450 mg, 1.411 mmol) in dry DMF (1 mL) was added 8-1a (1 M). The reaction mixture reacted at 75 degrees for 3 hours. The reaction solution was diluted with H.sub.2O (50 mL) under ice water bath, then extracted with PE/EA=1/1(50 mL3). The combined organics were washed with saturated salt water, dried with Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (petroleum ether) to give 8-2 (300 mg, yield: 80%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 2.20-2.16 (m, 2H), 1.93 (t, J=4.0 Hz, 1H), 1.54-1.49 (m, 2H), 1.40-1.26 (m, 28H), 0.88 (t, J=8.0 Hz, 3H).

(164) Step 2: To a solution of 4-8 (450 mg, 1.3 mmol) in DMF (10 mL) was added NaN.sub.3 (169 mg, 2.6 mmol), the mixture was stirred at 80 C. for 16 h. The mixture was diluted with water (100 mL) and extracted with EA (50 mL*3). The combined organic layer was washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (PE/EA=1:1) to afford 8-3 (450 mg) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.00 (d, J=8.0 Hz, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.51 (t, J=7.6 Hz, 1H), 7.34 (t, J=7.6 Hz, 1H), 4.90 (s, 2H), 4.27 (t, J=6.4 Hz, 2H), 3.41 (t, J=6.4 Hz, 2H), 2.93-2.79 (m, 2H), 2.08-1.93 (m, 2H), 1.93-1.63 (m, 4H), 1.47-1.38 (m, 2H), 0.94 (t, J=7.2 Hz, 3H). MS m/z (ESI): 354.2 [M+H].sup.+.

(165) Step 3: To a solution of 8-3 (200 mg, 0.567 mmol) in THF/H.sub.2O=1:1 (20 mL) were added CuSO.sub.4 (18 mg, 0.113 mmol) and Sodium L-Ascorbate (45 mg, 0.227 mmol), the mixture was stirred at RT for 1 h. The mixture was diluted with water (20 mL) and extracted with EA (30 mL*2). The combined organic layer was washed with brine (80 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by prep-HPLC (HCl in water/MeCN) to give the desired product (HCl salt), which was stirred in sat. sodium carbonate overnight to give the free product 3 (103.44 mg, yield: 29.4%) as a white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.04 (d, J=7.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (t, J=7.2 Hz, 1H), 7.37 (t, J=7.2 Hz, 1H), 7.30 (s, 1H), 5.88 (s, 2H), 4.48 (t, J=6.4 Hz, 2H), 4.31 (t, J=6.0 Hz, 2H), 2.92 (t, J=7.6 Hz, 2H), 2.73 (t, J=7.6 Hz, 2H), 2.27-2.21 (m, 2H), 2.02-1.98 (m, 2H), 1.92-1.85 (m, 2H), 1.70-1.65 (m, 2H), 1.52-1.46 (m, 2H), 1.33-1.25 (m, 28H), 0.99 (t, J=7.2 Hz, 3H), 0.88 (t, J=6.4 Hz, 3H). MS m/z (ESI): 618.5 [M+H].sup.+.

Example 9. 2-butyl-1-(4-(5-heptadecyl-1,3,4-oxadiazol-2-yl)butoxy)-1H-imidazo[4,5-c]quinolin-4-amine (9)

(166) ##STR00053##

(167) ##STR00054##

(168) Step 1: To a solution of 4-8 (2.7 g, 7.78 mmol) in DMSO (60 mL) was added KCN (1.01 g, 15.6 mmol), the mixture was stirred at 80 C. for 16 h. The mixture was diluted with water (100 mL) and extracted with EA (100 mL*3). The combined organic layer was washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (PE/EA=0:1) to afford 9-1 (2.6 g, yield: 87%) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.04 (dd, J=8.0, 1.2 Hz, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.55 (dd, J=11.2, 4.4 Hz, 1H), 7.37 (s, 1H), 6.04 (s, 2H), 4.35 (d, J=8.0 Hz, 2H), 2.93 (d, J=8.0 Hz, 2H), 2.56 (d, J=7.2 Hz, 2H), 2.20-2.14 (m, 2H), 2.03 (dd, J=10.0, 5.8 Hz, 2H), 1.89 (d, J=7.6 Hz, 2H), 1.50 (dd, J=15.0, 7.5 Hz, 2H), 1.01 (t, J=7.2 Hz, 3H).

(169) Step 2: A solution of 9-1 (800 mg, 2.37 mmol) in con.HCl (2 mL) was stirred at 100 C. for 4 h. After cooling to rt, the mixture was diluted with water (5 mL) and extracted with EA (20 mL*8). The combined organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give 9-2 (650 mg, yield: 77%) as a yellow solid. MS m/z (ESI): 357.2 [M+H].sup.+

(170) Step 3: To a solution of 9-2 (260 mg, 0.73 mmol) in DCM/DMF=4:1 (25 mL) were added stearohydrazide (435 mg, 1.46 mmol), EDCI (277 mg, 1.46 mmol) and HOBt (197 mg, 1.46 mmol), the mixture was stirred at RT overnight. The mixture was concentrated in vacuo, the residue was purified by prep-HPLC (HCl) to give compound 9-3 (120 mg, yield: 25.8%) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.97 (d, J=8.0 Hz, 1H), 7.80 (d, J=8.2 Hz, 1H), 7.53 (d, J=7.0 Hz, 1H), 7.31 (s, 1H), 4.23 (t, J=4.4 Hz, 2H), 2.94-2.83 (m, 2H), 2.42 (t, J=6.2 Hz, 2H), 2.30-2.22 (m, 2H), 2.07-1.97 (m, 4H), 1.87 (dd, J=15.4, 7.5 Hz, 3H), 1.69 (d, J=6.8 Hz, 4H), 1.53-1.47 (m, 3H), 1.37-1.21 (m, 36H), 1.00 (t, J=7.2 Hz, 3H), 0.88 (t, J=6.8 Hz, 3H). MS m/z (ESI): 637.5 [M+H].sup.+.

(171) Step 4: A solution of 9-3 (120 mg, 0.189 mmol) in POCl.sub.3 (2 mL) was stirred at 50 C. for 3 h. After cooling to RT, the mixture was poured into sodium carbonate solution (30 mL) and extracted with EA (30 mL*2). The combined organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by prep-TLC (DCM/MeOH=10:1) to give compound 9 (34.32 mg, yield: 29.3%) as a light-yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.07 (d, J=8.0 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (t, J=7.6 Hz, 1H), 7.38 (t, J=7.6 Hz, 1H), 6.07 (s, 2H), 4.35 (t, J=5.6 Hz, 2H), 2.97 (dt, J=15.4, 7.3 Hz, 4H), 2.83 (t, J=7.6 Hz, 2H), 2.14 (dd, J=16.1, 2.9 Hz, 4H), 1.89 (dt, J=15.3, 7.6 Hz, 2H), 1.79 (dd, J=14.8, 7.5 Hz, 2H), 1.49 (dd, J=15.0, 7.4 Hz, 2H), 1.31-1.25 (m, 28H), 1.00 (t, J=7.2 Hz, 3H), 0.88 (t, J=6.8 Hz, 3H). MS m/z (ESI): 619.5 [M+H].sup.+.

Example 10. 2-butyl-1-(4-(5-heptadecyl-1,2,4-oxadiazol-3-yl)butoxy)-1H-imidazo[4,5-c]quinolin-4-amine (10)

(172) ##STR00055##

(173) ##STR00056##

(174) Step 1: To a stirred solution of 9-1 (500 mg, 1.48 mmol) in EtOH (20 mL) under argon atmosphere were added hydroxylamine hydrochloride (512 mg, 7.42 mmol) and TEA (749 mg, 7.42 mmol), the mixture was stirred at RT for 16 h. The reaction was concentrated in vacuo and the residue was purified by column chromatography (DCM/MeOH=10:1 to 4:1) to give 10-2 (70 mg, yield: 12.8%) as a light green solid. .sup.1H NMR (400 MHz, DMSO-d6) 8.08 (d, J=8.0 Hz, 1H), 7.66 (d, J=8.0 Hz, 1H), 7.57-7.52 (m, 1H), 7.41-7.37 (m, 1H), 4.38-4.33 (m, 2H), 2.98-2.94 (m, 2H), 2.70-2.64 (m, 2H), 2.35-2.29 (m, 2H), 1.87-1.77 (m, 4H), 1.46-1.41 (m, 2H), 0.95 (t, J=7.2 Hz, 3H). MS m/z (ESI): 371.3 [M+H].sup.+.

(175) Step 2: To a solution of 10-2 (70 mg, 0.189 mmol) and stearic acid (107 mg, 0.378 mmol) in DMF/DCM=1:4 (10 mL) was added N,N-dicyclohexylcarbodiimide (DCC, 77.9 mg, 0.378 mmol), the mixture was stirred at RT for 20 h. The mixture was concentrated in vacuo to remove DCM and the residue was purified by prep-HPLC (FA) to give 10-3 (17 mg, yield: 14.2%) as a yellow solid. MS m/z (ESI): 637.5 [M+H].sup.+.

(176) Step 3: A solution of 10-3 (17 mg, 0.026 mmol) in DMF (2 mL) was stirred at 120 C. for 4 h. The mixture was purified by prep-HPLC (HCl) to give compound 10 (2.60 mg, yield: 15.7%) as a light solid. .sup.1H NMR (400 MHz, CD3OD) 8.30 (d, J=8.0 Hz, 1H), 7.77 (d, J=4.0 Hz, 2H), 7.66 (dd, J=8.0, 4.0 Hz, 1H), 4.51-4.47 (m, 2H), 3.08-3.03 (m, 2H), 2.95-2.86 (m, 4H), 2.09-2.07 (m, 2H), 2.06-1.98 (m, 2H), 1.97-1.91 (m, 2H), 1.83-1.77 (m, 2H), 1.64-1.57 (m, 2H), 1.54-1.50 (m, 2H), 1.30-1.23 (m, 26H), 1.02 (t, J=7.2 Hz, 3H), 0.88 (d, J=7.2 Hz, 3H). MS m/z (ESI): 619.9 [M+H].sup.+.

Example 11. 2-butyl-1-(4-(3-heptadecyl-1,2,4-oxadiazol-5-yl)butoxy)-1H-imidazo[4,5-c]quinolin-4-amine (11)

(177) ##STR00057##

(178) ##STR00058##

(179) Step 1: To a solution of NH.sub.2OHHCl (417 mg, 6.04 mmol) in isopropanol (15 mL) was added NaHCO.sub.3(761 mg, 9.06 mmol), the mixture was stirred at RT for 30 min, then 11-1 (800 mg, 3.02 mmol) was added to the mixture and stirred at 80 C. overnight. The mixture was filtered to give 11-2 (600 mg, yield: 66.7%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) 8.65 (s, 1H), 5.27 (s, 2H), 1.94-1.90 (m, 2H), 1.44 (d, J=6.4 Hz, 2H), 1.23 (s, 28H), 0.85 (t, J=6.8 Hz, 3H).

(180) Step 2: To a solution of 11-2 (146 mg, 0.504 mmol) and 9-10 (150 mg, 0.42 mmol) in dioxane/DCM=1:1 (16 mL) was added DCC (173 mg, 0.84 mmol), the mixture was stirred at RT for 2 h. The mixture was concentrated in vacuo, the residue was purified by prep-TLC (DCM/MeOH=10:1) to give 13-3 (180 mg, yield: 67.4%) as a yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.09 (d, J=8.0 Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.61-7.57 (m, 1H), 7.44-7.40 (m, 1H), 4.34 (t, J=6.0 Hz, 2H), 2.98-2.92 (m, 2H), 2.58 (t, J=6.8 Hz, 2H), 2.28-2.22 (m, 2H), 2.09-2.01 (m, 4H), 1.90 (d, J=7.7 Hz, 4H), 1.63-1.59 (m, 2H), 1.53-1.48 (m, 2H), 1.37-1.24 (m, 26H), 1.00 (t, J=7.2 Hz, 3H), 0.88 (t, J=6.8 Hz, 3H).

(181) Step 3: A solution of 13-3 (180 mg, 0.283 mmol) in DMF (5 mL) was stirred at 120 C. for 4 h. The mixture was purified by prep-HPLC (HCl) to give compound 11 (63.61 mg, yield: 36%) as a light-yellow solid (HCl salt). .sup.1H NMR (400 MHz, CDCl.sub.3) 15.09 (s, 1H), 9.09 (s, 1H), 8.12 (d, J=8.0 Hz, 1H), 8.00 (d, J=8.0 Hz, 1H), 7.71 (d, J=7.2 Hz, 1H), 7.57 (d, J=7.2 Hz, 1H), 7.00 (s, 1H), 4.43 (d, J=2.4 Hz, 2H), 3.10-2.93 (m, 4H), 2.76-2.68 (m, 2H), 2.20-2.08 (m, 4H), 2.04-1.91 (m, 4H), 1.76 (dd, J=14.6, 7.6 Hz, 2H), 1.52 (dt, J=14.3, 7.2 Hz, 2H), 1.39-1.22 (m, 26H), 1.02 (t, J=7.3 Hz, 3H), 0.88 (t, J=6.8 Hz, 3H). MS m/z (ESI): 619.5 [M+H].sup.+.

Example 12. 3-(4-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yloxy)butylamino)-4-((3-(dimethylamino)propyl)(heptadecyl)amino)cyclobut-3-ene-1,2-dione (12)

(182) ##STR00059##

(183) ##STR00060##

(184) Step 1: A solution of 5-1 (0.3 g, 1.18 mmol) and 4-8a (120.47 mg, 1.18 mmol) in MeOH (20 mL) was stirred at 30 C. for 3 hours, NaBH.sub.4 (44.61 mg, 1.18 mmol) was added to the mixture and stirred at 30 C. for another 16 hours. LCMS showed product was formed. The mixture was diluted with water (20 mL), extracted with EtOAc (40 mL*2). The combined organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give 12-1 (0.3 g, 75% yield) as a light yellow oil, which was confirmed by HNMR. .sup.1H NMR (400 MHz, CDCl3) 2.65 (t, J=6.8 Hz, 2H), 2.59 (t, J=7.2 Hz, 2H), 2.33-2.29 (m, 2H), 2.22 (s, 6H), 1.70-1.65 (m, 2H), 1.50-1.44 (m, 1H), 1.32-1.20 (m, 28H), 0.87 (d, J=6.8 Hz, 3H).

(185) Step 2: A solution of 12-1 (0.3 g, 0.880 mmol) and 12-2 (125.2 mg, 0.880 mmol) in MeOH (20 mL) was stirred at 30 C. for 4 hours. TLC (DCM/MeOH=10:1) showed the reaction was completed. The mixture was concentrated in vacuo, the residue was purified by column chromatography (DCM/MeOH=10:1) to give compound 12-3 (0.28 g, 71% yield) as a light yellow oil. MS m/z (ESI) 451.2 [M+H].sup.+.

(186) Step 3: To a solution of 12-3(42.0 mg, 0.093 mmol) and compound A (30.5 mg, 0.093 mmol) in MeOH (4 mL) was added DIEA (24.1 mg, 0.186 mmol, 32.46 uL), the mixture was stirred at room temperature for 16 hours. LCMS showed the reaction was completed. The mixture was concentrated in vacuo, the residue was purified by prep-HPLC (0.05% HCl) to give Compound 12 as HCl salt (37 mg, 53% yield) as a white solid. .sup.1H NMR (400 MHz, MeOD) 8.19 (d, J=7.6 Hz, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.68 (t, J=7.2 Hz, 1H), 7.58 (t, J=7.2 Hz, 1H), 4.50 (br.s, 2H), 3.86-3.55 (m, 6H), 3.29-3.22 (m, 2H), 3.04 (t, J=7.2 Hz, 2H), 2.94 (s, 6H), 2.14-2.11 (m, 4H), 2.01-1.88 (m, 4H), 1.67 (br.s, 2H), 1.57-1.48 (m, 2H), 1.34-1.19 (m, 28H), 1.02 (t, J=7.2 Hz, 3H), 0.87 (t, J=6.8 Hz, 3H). MS m/z (ESI) 746.5 [M+H].sup.+.

Example 13. TLR7/8 agonists induce IFN-gamma and TNF-alpha release in human PBMC

(187) The system is used to assess the cytokines release. Activity is based on the measurement of interferon-gamma (IFN-) and tumor necrosis factor-alpha (TNF-) secreted into culture media.

(188) Isolation of PBMCs

(189) Fresh human blood was diluted with the same volume of PBS, 15 mL Lymphoprep was added into a Sepmate tube, then 30 mL diluted blood was added on the top gently without disturbing the interface.

(190) The Sepmate tube was centrifuged for 25 min at 1000g at RT with brake off.

(191) The buffy coat containing peripheral blood mononuclear cells (PBMCs) was collected from Sepmate tube and transferred into a new tube, and the cells were washed with 40 mL PBS twice and centrifuged at 350g for 5 min.

(192) PBMCs were resuspended in complete culture medium at a density of 2E6/mL.

(193) Compound Preparation

(194) The compounds are solubilized in dimethyl sulfoxide (DMSO) and diluted into indicated concentration with complete culture medium.

(195) The compounds are tested at final concentrations 100 M, 33.3 M, 11.1 M, 3.7 M, 1.23 M, 0.41 M, 0.137 M, 0.0457 M and 0. 0152 M.

(196) Incubation

(197) 2*10^5 PBMCs (in 100 L) were added to each well of 96-well flat bottom plate.

(198) 2 final concentration of 3-fold serial diluted compounds (in 100 L) were added to indicated wells and final volume was 200 L.

(199) The plate was covered with sterile lids, mixed gently and then incubated for 24 h at 37 C./5% CO.sub.2 incubator.

(200) Separation Supernatant

(201) Following incubation, the plates was centrifuged for 5 min. at 400g. The cell-free culture Supernatant was removed into a non-sterile polypropylene plate. Samples are maintained at 80 C. until analysis. The samples were analyzed for TNF- and IFN- by ELISA according to the direction.

(202) TNF- and IFN- were analyzed by ELISA. IFN- concentration was determined by ELISA using a Human IFN- ELISA Kit from R&D Systems (Catalog #41100-2) and read on VICTOR Nivo from PerkinElmer. Results were expressed in pg/mL. TNF- concentration was determined by ELISA using a Human TNF-alpha ELISA MAX Deluxe from BioLegend (Catalog #430205) and read on VICTOR Nivo from PerkinElmer. Results were expressed in pg/mL.

(203) The data was analyzed to determine the minimum effective concentration (MEC) for each compound at which induction of a particular cytokine was observed in the assay. Specifically, the MEC of each compound (micromolar) was determined as the lowest concentration of the compound that induced a measured cytokine response at a level (pictograms/mL) that was at least 2 greater than that observed with the negative control wells. The results are presented in Table 2.

(204) TABLE-US-00002 TABLE 2 MEC to induce cytokine (micromolar) Compound IFN-alpha TNF-alpha Example 1 >33.3 11.1 Example 2 0.0152 0.0457 Example 3 0.137 0.137 Example 4 33.3 33.3 Example 5 1.23 11.1 Example 6 >100 >100 Example 7 0.0152 0.41 Example 8 0.137 0.41 Example 8 3.7 33.3 Example 10 11.1 3.7 Example 11 11.1 33.3 Example 12 0.0152 0.41
Exemplary Pharmaceutical Preparations

(205) (A) Injection vials: A solution of 100 g of an active ingredient according to the invention and 5 g of disodium hydrogen phosphate in 3 L of distilled water is adjusted to pH 6.5 using 2 N hydrochloric acid, sterile filtered, transferred into injection vials, is lyophilized under sterile conditions and is sealed under sterile conditions. Each injection vial contains 5 mg of active ingredient.

(206) (B) Suppositories: A mixture of 20 g of an active ingredient according to the invention is melted with 100 g of soy lecithin and 1400 g of cocoa butter, is poured into moulds and is allowed to cool. Each suppository contains 20 mg of active ingredient.

(207) (C) Solution: A solution is prepared from 1 g of an active ingredient according to the invention, 9.38 g of NaH.sub.2PO.sub.4.2 H.sub.2O, 28.48 g of Na.sub.2HPO.sub.4.12 H.sub.2O and 0.1 g of benzalkonium chloride in 940 ml of bidistilled water. The pH is adjusted to 6.8, and the solution is made up to 11 and sterilized by irradiation. This solution could be used in the form of eye drops.

(208) (D) Ointment: 500 mg of an active ingredient according to the invention is mixed with 99.5 g of Vaseline under aseptic conditions.

(209) (E) Tablets: A mixture of 1 kg of an active ingredient according to the invention, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is pressed to give tablets in a conventional manner in such a way that each tablet contains 10 mg of active ingredient.

(210) (F) Coated tablets: Tablets are pressed analogously to Example E and subsequently are coated in a conventional manner with a coating of sucrose, potato starch, talc, tragacanth and dye.

(211) (G) Capsules: 2 kg of an active ingredient according to the invention are introduced into hard gelatin capsules in a conventional manner in such a way that each capsule contains 20 mg of the active ingredient.

(212) (H) Ampoules: A solution of 1 kg of an active ingredient according to the invention in 60 l of bidistilled water is sterile filtered, transferred into ampoules, is lyophilized under sterile conditions and is sealed under sterile conditions. Each ampoule contains 10 mg of active ingredient.

(213) (I) Inhalation spray: 14 g of an active ingredient according to the invention are dissolved in 101 of isotonic NaCl solution, and the solution is transferred into commercially available spray containers with a pump mechanism. The solution could be sprayed into the mouth or nose. One spray shot (about 0.1 ml) corresponds to a dose of about 0.14 mg.

(214) Applicant's disclosure is described herein in preferred embodiments with reference to the Figures, in which like numbers represent the same or similar elements. Reference throughout this specification to one embodiment, an embodiment, or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases in one embodiment, in an embodiment, and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

(215) The described features, structures, or characteristics of Applicant's disclosure may be combined in any suitable manner in one or more embodiments. In the description, herein, numerous specific details are recited to provide a thorough understanding of embodiments of the invention. One skilled in the relevant art will recognize, however, that Applicant's composition and/or method may be practiced without one or more of the specific details, or with other methods, components, materials, and so forth. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosure.

(216) In this specification and the appended claims, the singular forms a, an, and the include plural reference, unless the context clearly dictates otherwise.

(217) Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Methods recited herein may be carried out in any order that is logically possible, in addition to a particular order disclosed.

INCORPORATION BY REFERENCE

(218) References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made in this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material. In the event of a conflict, the conflict is to be resolved in favor of the present disclosure as the preferred disclosure.

EQUIVALENTS

(219) The representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples and the references to the scientific and patent literature included herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.