NOVEL NON-OPIOID ANTI-PAIN MEDICATION
20220185855 · 2022-06-16
Inventors
- Fernanda Laezza (Galveston, TX, US)
- Jia Zhou (Galveston, TX)
- Jin Mo Chung (Galveston, TX, US)
- Plngyuan Wang (Galveston, TX, US)
- Jun-Ho La (Galveston, TX, US)
- Oluwarotimi Folorunso (Galveston, TX, US)
- Aditya Singh (Galveston, TX, US)
Cpc classification
C07K5/1024
CHEMISTRY; METALLURGY
International classification
Abstract
The present invention relates to novel small molecules Formula I and pharmaceutically acceptable salts thereof as well as the preparation and the use thereof to inhibit FGF13-1b.
##STR00001##
Claims
1. A compound of Formula I or a pharmaceutically acceptable salt thereof, wherein: ##STR00018## R.sup.1 is H, alkyl, cycloalkyl, aryl, heteroaryl, R.sup.4CO—, R.sup.5NHCO—, R.sup.6OCO—, R.sup.7SO.sub.2—, or Fmoc; where R.sup.4, R.sup.5, R.sup.6 and R.sup.7 are independently selected from the group consisting of alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, adamantyl, and benzyl, wherein R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is optionally substituted with one or more chosen substituents chosen from —OH, —CN, —NH.sub.2, and halogen; R.sup.2 is H, alkyl, aryl, heteroaryl, cycloalkyl, or Boc; R.sup.3 is OH, alkoxy, allyloxy, —NR.sup.8R.sup.9; wherein R.sup.8 and R.sup.9 are independently selected from the group consisting of H, alkyl, aryl, and heteroaryl; or R.sup.8 and R.sup.9 are optionally joined to form a N-containing heterocycle with 1-3 heteroatoms; and wherein R.sup.1 and R.sup.2 are not both hydrogen.
2. The compound of claim 1, wherein R.sup.1 is H.
3. The compound of claim 1, wherein R.sup.1 is R.sup.4CO— and R.sup.4 is alkyl, cycloalkyl, alkenyl, aryl, or Fmoc.
4. The compound of claim 1, wherein R.sup.1 is R.sup.5NHCO— and R.sup.5 is adamantyl.
5. The compound of claim 1, wherein R.sup.1 is R.sup.6OCO— and R.sup.6 is benzyl or substituted benzyl.
6. The compound of claim 1, wherein R.sup.1 is R.sup.7SO.sub.2— and R.sup.7 is 4-fluorophenyl.
7. The compound of claim 1, wherein R.sup.2 is Boc.
8. The compound of claim 1, wherein R.sup.3 is OH, —OMe, or allyloxy.
9. The compound of claim 1, wherein R.sup.3 is —NR.sup.8R.sup.9 and R.sup.8 and R.sup.9 is chosen from H and any of the following substituents: ##STR00019##
10. The compound of claim 1, wherein the compound is chosen from: ##STR00020## ##STR00021## ##STR00022## ##STR00023##
11. The compound of claim 1, wherein R.sup.1 is R.sup.6OCO—, where R.sup.6 is selected from the group consisting of alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, and benzyl, wherein R.sup.6 is optionally substituted with one or more chosen substituents chosen from —OH, —CN, —NH.sub.2, and halogen; R.sup.2 is alkyl; R.sup.3 is OH, —OMe, allyloxy, or -NR.sup.8R.sup.9 and R.sup.8 and R.sup.9 is chosen from H or any of the following substituents: ##STR00024##
12. The compound of claim 11, wherein R.sup.2 is t-butyl.
13. The compound of claim 11, wherein R.sup.6 is benzyl or substituted benzyl.
14. The compound of claim 1, wherein R.sup.4CO— and R.sup.4 is 4-fluorophenyl.
15. The compound of claim 1, wherein R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently methyl or nC.sub.2-C.sub.9 alkyl.
16. The compound of claim 1, wherein R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently n-nonyl.
17. The compound of claim 1, wherein R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently 4-fluorophenyl.
18. A method of inhibiting FGF13-1b comprising contacting one or more cells with one or more compounds of claim 1 or pharmaceutically acceptable salt thereof.
19. A method of treating or alleviating pain by modulating FGF13-1b comprising administering one or more compounds of claim 1 or pharmaceutically acceptable salt thereof to a subject.
Description
BRIEF DESCRIPTION OF THE FIGURES
[0016]
[0017]
[0018]
[0019]
[0020]
[0021]
[0022]
[0023]
[0024]
[0025]
[0026]
[0027]
[0028]
[0029] (a) PW164 has moderate oral bioavailability and moderate oral half-life. PW164 has no hERG and CYP450s liabilities. PW164 has high human plasma metabolic stability and human whole blood stability; and
[0030] (b) PW164 cannot penetrate BBB. This might be favorable for peripheral therapeutics with the advantage of no brain-associated side toxicity effects.
DETAILED DESCRIPTION
1.0. Definitions
1.0. DEFINITIONS
[0031] For the purposes of promoting an understanding of the principles of the invention, reference will now be made to certain embodiments and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, and alterations and modifications in the illustrated invention, and further applications of the principles of the invention as illustrated therein are herein contemplated as would normally occur to one skilled in the art to which the invention relates.
[0032] 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 to which this invention pertains.
[0033] For the purpose of interpreting this specification, the following definitions will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. In the event that any definition set forth below conflicts with the usage of that word in any other document, including any document incorporated herein by reference, the definition set forth below shall always control for purposes of interpreting this specification and its associated claims unless a contrary meaning is clearly intended (for example in the document where the term is originally used).
[0034] The use of “or” means “and/or” unless stated otherwise.
[0035] The use of “a” herein means “one or more” unless stated otherwise or where the use of “one or more” is clearly inappropriate.
[0036] The use of “comprise,” “comprises,” “comprising,” “include,” “includes,” and “including” are interchangeable and not intended to be limiting. Furthermore, where the description of one or more embodiments uses the term “comprising,” those skilled in the art would understand that, in some specific instances, the embodiment or embodiments can be alternatively described using the language “consisting essentially of” and/or “consisting of.”
[0037] As used herein, the term “about” refers to a ±10% variation from the nominal value. It is to be understood that such a variation is always included in any given value provided herein, whether or not it is specifically referred to.
[0038] The term “pharmaceutically acceptable salt” refers to those salts of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, and the like. As used herein, the term “pharmaceutically acceptable salt” may include acetate, hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactobionate and laurylsulphonate salts, and the like. (See S. M. Barge et al., “Pharmaceutical Salts,” J. Pharm. Sci., 66:1-19 (1977), which is incorporated herein by reference in its entirety, for further examples of pharmaceutically acceptable salt).
[0039] The term “HBTU” refers to 3-[Bis(dimethylamino)methyliumyl]-3H -benzotriazol-1-oxide hexafluorophosphate (also known as 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate).
[0040] The term “HOBt” refers the following structure, known as 1-hydroxybenzotriazole, (including hydrates and polymorphs, thereof):
##STR00003##
[0041] The term “DIEA” refers to N,N-Diisopropylethylamine (also known as Hünig' s base, DIPEA, and ethyldiisopropylamine).
[0042] The term “DCM” refers to dichloromethane (also known as methylene chloride).
[0043] The term “EDCI” refers to 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC, EDAC or EDCI).
[0044] The term “TFA” refers to trifluoroacetic acid.
[0045] The term “rt” refers to room temperature.
[0046] The term “alkyl” as used herein by itself or as part of another group refers to both straight and branched chain radicals, and cyclic alkyl groups. In one embodiment, the alkyl group has 1-12 carbons. In another embodiment, the alkyl group has 1-7 carbons. In another embodiment, the alkyl group has 1-6 carbons. In another embodiment, the alkyl group has 1-4 carbons. The term “alkyl” may include methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, and dodecyl.
[0047] The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a linear or branched chain having at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, S, P, and Si. In certain embodiments, the heteroatoms are selected from the group consisting of O, and N. The heteroatom(s) may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Up to two heteroatoms may be consecutive.
[0048] The term “alkylene” as used herein refers to straight and branched chain alkyl linking groups, i.e., an alkyl group that links one group to another group in a molecule. In some embodiments, the term “alkylene” may include —(CH.sub.2).sub.n— where n is 2-8.
[0049] The term “aryl” means a polyunsaturated hydrocarbon substituent. Aryl groups can be monocyclic or polycyclic (e.g., 2 to 3 rings that are fused together or linked covalently). Non-limiting examples of aryl and heteroaryl rings are phenyl, naphthyl, pyranyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyridinyl, furanyl, thiophenyl, thiazolyl, imidazolyl, isoxazolyl, and the like.
[0050] The term “heteroaryl” as used herein refers to groups having 5 to 14 ring atoms; 6, 10 or 14 7n-electrons shared in a cyclic array; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur heteroatoms. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Especially preferred heteroaryl groups include 1,2,3-triazole, 1,2,4-triazole, 5-amino 1,2,4-triazole, imidazole, oxazole, isoxazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 3-amino-1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, pyridine, 2-aminopyridine, 4-aminopyridine, 2-aminoimidazoline, and 4-aminoimidazoline.
[0051] An “amino” group refers to an —NH.sub.2 group.
[0052] An “amido” group refers to an —CONH.sub.2 group. An alkylamido group refers to an —CONHR group wherein R is as defined above. A dialkylamido group refers to an —CONRR′ group wherein R and R′ are as defined above.
[0053] The term “halogen” or “halo” as used herein by itself or as part of another group refers to chlorine, bromine, fluorine or iodine.
[0054] The term “hydroxy” or “hydroxyl” as used herein by itself or as part of another group refers to an —OH group.
[0055] An “alkoxy” group refers to an -O-alkyl group wherein “alkyl” is as defined above. In one embodiment, the alkyl group has 1-12 carbons. In another embodiment, the alkyl group has 1-7 carbons. In a further embodiment, the alkyl group has 1-6 carbons. In another embodiment, the alkyl group has 1-4 carbons.
[0056] A “thio” group refers to an —SH group.
[0057] An “alkylthio” group refers to an —SR group wherein R is alkyl as defined above.
[0058] The term “heterocycle” or “heterocyclic ring”, as used herein except where noted, represents a stable 5- to 7-membered monocyclic-, or stable 7- to 11-membered bicyclic heterocyclic ring system, any ring of which may be saturated or unsaturated, and which consists of carbon atoms and from one to three heteroatoms selected from the group consisting of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. Rings may contain one oxygen or sulfur, one to three nitrogen atoms, or one oxygen or sulfur combined with one or two nitrogen atoms. The heterocyclic ring may be attached at any heteroatom or carbon atom that results in the creation of a stable structure.
[0059] The term “alkylamino” as used herein by itself or as part of another group refers to an amino group which is substituted with one alkyl group having from 1 to 6 carbon atoms. The term “dialkylamino” as used herein by itself or as part of another group refers to an amino group which is substituted with two alkyl groups, each having from 1 to 6 carbon atoms.
[0060] The term “arylamine” or “arylamino” as used herein by itself or as part of another group refers to an amino group which is substituted with an aryl group, as defined above.
[0061] As used herein, the term “arylalkyl” denotes an alkyl group substituted with an aryl group, for example, Ph-CH.sub.2— etc.
[0062] Various groups are described herein as substituted or unsubstituted (i.e., optionally substituted). Optionally substituted groups may include one or more substituents independently selected from: halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, oxo, carbamoyl, alkyl, heteroalkyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkyl sulfonyl, aryl sulfonyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. In certain aspects the optional substituents may be further substituted with one or more substituents independently selected from: halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl (—C(O)NR.sub.2), unsubstituted alkyl, unsubstituted heteroalkyl, alkoxy, alkylthio, alkylamino, (alkyl).sub.2amino, alkylsulfinyl, alkyl sulfonyl, aryl sulfonyl, unsubstituted cycloalkyl, unsubstituted heterocyclyl, unsubstituted aryl, or unsubstituted heteroaryl. Exemplary optional substituents include, but are not limited to: —OH, oxo (═O), —Cl, —F, Br, C.sub.1-4alkyl, phenyl, benzyl, —NH.sub.2, —NH(C.sub.1-4alkyl), —N(C1-4alkyl).sub.2, —NO2, —S(C.sub.1-4alkyl), —SO.sub.2(C.sub.1-4alkyl), —CO.sub.2(C.sub.1-4alkyl), and —O(C.sub.1-4alkyl).
[0063] PW0164 is used throughout this description interchangeably with PW164 to refer to:
##STR00004##
[0064] The term “subject” as used herein refers to an animal, preferably a mammal, most preferably a human, who has been the object of treatment, observation or experiment. Preferably, the subject has experienced and/or exhibited at least one symptom of the disease or disorder to be treated and/or prevented. Further, a subject may not have exhibited any symptoms of the disorder, disease or condition to be treated and/prevented, but has been deemed by a physician, clinician or other medical professional to be at risk for developing said disorder, disease or condition.
[0065] The terms “treating,” “treatment” and the like as used herein includes the management and care of a subject (preferably a mammal, more preferably a human) for the purpose of combating a disease, condition, or disorder and includes the administration of a compound of the present disclosure to prevent the onset of the symptoms or complications, alleviate the symptoms or complications, or eliminate the disease, condition, or disorder.
[0066] It is to be understood that both the foregoing descriptions are exemplary, and thus do not restrict the scope of the invention.
2.0. COMPOUNDS
[0067] The present invention provides a compound of the Formula I and pharmaceutically acceptable salts thereof, wherein:
##STR00005##
[0068] R.sup.1 is H, alkyl, cycloalkyl, aryl, heteroaryl, R.sup.4CO—, R.sup.5NHCO—, R.sup.6OCO—, R.sup.7SO.sub.2—, or Fmoc-;
[0069] where R.sup.4, R.sup.5, R.sup.6 and R.sup.7 are independently selected from the group consisting of alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, adamantyl, and benzyl, wherein R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is optionally substituted with one or more chosen substituents chosen from —OH, —CN, —NH.sub.2, and halogen;
[0070] R.sup.2 is H, alkyl, aryl, heteroaryl, cycloalkyl, or Boc;
[0071] R.sup.3 is OH, alkoxy, allyloxy, —NR.sup.8R.sup.9; wherein R.sup.8 and R.sup.9 are independently selected from the group consisting of H, alkyl, aryl, and heteroaryl; or R.sup.8 and R.sup.9 are optionally joined to form a N-containing heterocycle with 1-3 heteroatoms; and
[0072] wherein R.sup.1 and R.sup.2 are not both hydrogen.
[0073] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.1 is H.
[0074] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.1 is R.sup.4CO— and R.sup.4 is alkyl, cycloalkyl, alkenyl, aryl, or Fmoc.
[0075] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.1 is R.sup.5NHCO— and R.sup.5 is adamantyl.
[0076] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.1 is R.sup.6OCO— and R.sup.6 is benzyl or substituted benzyl.
[0077] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, wherein R.sup.1 is R.sup.7SO.sub.2— and R.sup.7 is 4-fluorophenyl.
[0078] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.2 is Boc.
[0079] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.3 is OH, —OMe, or allyloxy.
[0080] In some embodiments of Formula I and pharmaceutically acceptable salts thereof, R.sup.3 is —NR.sup.8R.sup.9 and R.sup.8 and R.sup.9 is chosen from H and any of the following substituents:
##STR00006##
[0081] In some embodiments, Formula I is chosen from:
##STR00007## ##STR00008## ##STR00009## ##STR00010##
[0082] In some embodiments of Formula I and pharmaceutically acceptable salts thereof:
R.sup.1 is R.sup.6OCO—;
R.sup.6 is selected from the group consisting of alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, and benzyl, wherein R.sup.6 is optionally substituted with one or more chosen substituents chosen from —OH, —CN, —NH.sub.2, and halogen;
R.sup.2 is alkyl; and
R.sup.3 is OH, —OMe, allyloxy, or —NR.sup.8R.sup.9 and R.sup.8 and R.sup.9 is chosen from H or any of the following substituents:
##STR00011##
[0083] In some embodiments, R.sup.2 is t-butyl.
[0084] In some embodiments, R.sup.1 is R.sup.4CO— wherein R.sup.4 is 4-fluorophenyl.
[0085] In some embodiments, R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently methyl or nC.sub.2-C.sub.9 alkyl.
[0086] In some embodiments, R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently n-nonyl.
[0087] In some embodiments, R.sup.4, R.sup.5, R.sup.6 and R.sup.7 is independently 4-fluorophenyl.
2.1. Synthesis of Compounds of the Invention
[0088] The description of preparation of certain compounds of the invention is meant to be exemplary of certain embodiments of the invention. The reagents and reactant used for synthetic conversions outlined herein and below is merely exemplary. The invention contemplates using the same or different reagents discussed herein to achieve preparation of the compounds of the invention.
[0089] Synthetic methods. The general methods used for the synthesis of various PW164 analogues (Series A-F) are depicted in Scheme 1 (below). Starting from commercially available Fmoc-Glu-OH 1, reaction with NH.sub.2-Val-OMe 2 in the presence of the Hunig's base DIPEA and catalyzed by HBTU, followed by deprotection the Fmoc group in the presence of the base DEA will give intermediate 3. Coupling of 3 with Cbz-Pro-Leu-OH 4 by HBTU and DIPEA will provide intermediate 5. Deprotection of 5 by Pd/C and H.sub.2 will lead to intermediate 6, which will react with various commercially available 7 to produce compounds of Series A. Series B and Series C will be generated in a similar fashion by reacting intermediate 6 with various commercially available 8 or 9. Intermediate 10 will be afforded by hydrolysis of the intermediate 5. 10 will react with various commercially available amines 11 to provide compounds of Series D. Hydrolyzing intermediate 5 using LiOH, followed by the treatment of 0.5% citric acid will afford intermediate 12 which will react with various commercially available alcohols 13 to provide compounds of Series E. Series F will be generated in a similar fashion by coupling of intermediate 12 with various commercially available amines 14.
##STR00012##
3.0. METHOD OF USE
[0090] Nav1.6 channels amplify response to painful stimuli (hyperalgesia) through sustained firing of sensory neurons. Studies have shown that two splice isoforms of fibroblast growth factor 13 (FGF13), an accessory protein that binds to the Nav1.6 intracellular C-terminal tail4-7 differentially regulate channel function with FGF13-1a conferring protection against radicular pain and FGF13-1b exacerbating it. The inventors surprisingly discovered certain compounds, as exemplified by PW164, inhibits FGF13-1b and acts as a FGF13-1a mimetic.
[0091] In some embodiments, the invention encompasses a method of inhibiting FGF13-1b comprising contacting one or more cells with one or more compounds of Formula I or pharmaceutically acceptable salts thereof.
[0092] In some embodiments, the invention encompasses treating or alleviating pain by inhibiting FGF13-1b comprising administering one or more compounds of Formula I or pharmaceutically acceptable salts thereof to a subject.
4.0 EXAMPLES
4.1 Evidence for Functional Activity of PW164.
[0093] FGF13 stands out among iFGFs as a potent, specific, and diverse modulator of Nav channels, especially Nav1.6. When expressed in recombinant cells, FGF13 can suppress or augment Nav1.6-encoded currents and channel availability, depending on its N-terminal spliced isoform with phenotypes that are reminiscent of other iFGFs in these cell lines. In animal models of pain, the FGF13-1b isoform is up-regulated, while FGF13-1a exerts a protective effect. Models in DRG attribute the beneficial effect of the FGF13-1a isoform in pain to a specific effect on Nav1.6-mediated persistent and resurgent currents that are thought to take part in DRG firing associated with pain.
[0094] Based on the success of other PPI regulators targeting ion channels we used the same rationale of selecting compounds that could act as a FGF13 isoform specific blocking FGF13-1b isoform effects mimicking FGF13-1a and select PW164 for functional assays. We used patch-clamp electrophysiology and begun functional validation of the lead compound using a panel of biophysical protocols targeting Nav channels (
[0095] These studies provide a strong premise to further optimize PW164 improving its potency and refine even further isoform selectivity. In addition, the strength of interaction and phenotypic regulation of Na+ currents are strong for the FGF13:Nav1.6 pair, but FGF13 binds and regulates other neuronal (Nav1.1 and Nav1.2) and cardiac16,30 Nav channels and other iFGFs, such as cardiac FGF14, modulate Nav1.6 currents. Thus, cross reactivity of optimized leads against FGF13:Nav channel isoform complexes other than Nav1.6, (e.g, Nav1.1, Nav1.2, and the predominant cardiac iFGF:Nav pair) should be ruled out. Our structure-function studies provide a strong premise for divergence at the iFGFs:Nav channel interface, providing us with further confidence in developing selective compounds that lack undesired effects on other targets. Because our strategy has proven successful in targeting PPI within Nav channels, PW164-derived analogues are expected to rapidly translate into leads with limited side effects on other Nav channel isoforms.
4.2 Evidence of Efficacy in Behavioral Models of Clinical Pain
[0096] Nav1.6 has been shown to play a crucial role in nociceptor excitation eliciting pain. For example, peripherally-injected scorpion toxin Cn2, a potent and selective Nav1.6 activator, produces spontaneous pain and mechanical hypersensitivity (i.e., enhanced pain from mechanical stimulation) whereas knockdown of Nav1.6 expression in DRG strongly inhibits mechanical hypersensitivity and hyper-excitability of DRG neurons in animal models of neuropathic and inflammatory pain. These studies suggest that PW164 and its analogues, as regulators of Nav1.6 function, will affect nociceptor excitability and the intensity of pain. In a preliminary in vivo study, we tested effects of PW164 on nociceptive behavior in normal and capsaicin-injected mice. Intradermal capsaicin injection is known to strongly excite nociceptors at the injection site and sensitizes peripheral nociceptors and the spinal nociceptive neural circuit, resulting in mechanical hypersensitivity at the capsaicin injection site.
[0097] In animals, hypersensitivity is manifested as increased withdrawals from von Frey filament stimulations of the injection site (
4.3 Synthetic Experimental Procedures of the Compounds
[0098] ##STR00013## ##STR00014## ##STR00015##
tert-Butyl (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-4(S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0160)
[0099] Fmoc-Glu(O.sup.tBu)-OH (1.3 g, 3 mmol) and H-Val-NH.sub.2⋅HCl (459 mg, 3 mmol) were dissolved in 20 mL DCM and the mixture solution was cooled to 0° C. with ice bath. HOBt (405 mg, 3 mmol), HBTU (2.3 g, 6 mmol) and DIPEA (2 mL, 12 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1) to obtain PW0160 (1.3 g, 77%) as a white solid. .sup.1NMR (300 MHz, Methanol-d.sub.4) δ 7.80 (d, J=7.5 Hz, 2H), 7.65 (d, J=6.1 Hz, 2H), 7.35 (dt, J=24.0, 7.7 Hz, 4H), 4.45-4.33 (m, 2H), 4.23 (dt, J=9.1, 4.5 Hz, 3H), 2.40-2.23 (m, 2H), 2.17-2.02 (m, 2H), 1.88 (s, 1H), 1.45 (d, J=2.3 Hz, 9H), 0.96 (td, J=7.0, 2.3 Hz, 6H). .sup.13C NMR (75 MHz, Methanol-d.sub.4) δ 172.74, 172.60, 157.05, 141.17, 127.37, 126.76, 124.78, 119.51, 80.44, 66.61, 58.15, 54.30, 47.52, 47.32, 47.03, 47.01, 46.75, 31.33, 30.63, 26.95, 18.34, 16.95.
Benzyl (S)-2-(((S)-1-(((S)-1-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-5-(tert-butoxy)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0161)
[0100] To a solution of DEA (10 mL) and MeCN (10 mL) was added PW0160 (1.05 g, 2 mmol) and the solution was stirred at room temperature for 1 h. After the reaction completed, the solution was concentrated. The crude residue was dissolved in 20 mL dry DCM and added Z-Pro-Leu-OH (724, 2 mmol). The mixture solution was cooled to 0° C. with ice bath. HOBt (270 mg, 2 mmol), HBTU (1.5 g, 4 mmol) and DIPEA (1.3 mL, 8 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1 to 20/1) to obtain PW0161 (0.96 g, 74%) as a white solid. .sup.1NMR (300 MHz, DMSO-d.sub.6) δ 8.16-7.83 (m, 2H), 7.53 (dd, J=22.4, 8.9 Hz, 1H), 7.43-7.21 (m, 6H), 7.05 (s, 1H), 5.19-4.87 (m, 2H), 4.43-4.06 (m, 4H), 3.59-3.32 (m, 2H), 2.18 (ddd, J=17.3, 11.3, 7.1 Hz, 3H), 1.97-1.46 (m, 8H), 1.38 (d, J=3.0 Hz, 9H), 0.97-0.68 (m, 12H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) δ 173.02, 172.54, 172.50, 172.40, 172.19, 171.11, 137.44, 128.82, 128.62, 128.22, 127.90, 127.28, 80.09, 66.37, 66.16, 60.29, 59.36, 57.68, 57.60, 52.20, 52.07, 51.65, 51.42, 47.57, 47.00, 41.05, 40.72, 31.62, 31.06, 30.99, 30.45, 28.20, 27.75, 27.60, 24.65, 24.57, 24.33, 23.39, 23.33, 22.02, 19.68, 18.11.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-4-methyl-2-((S)-pyrrolidine-2-carboxamido)pentanamido)-5-oxopentanoate (PW0165)
[0101] To a solution of PW0161 (1.1 g) in 100 mL MeOH, 10% Pd/C (100 mg) was added. Under H.sub.2, the mixture was allowed to stir at room temperature for 2 hrs. The solution was filtered, and the filtrate was concentrated to get the crude product. The residue was purified by silica gel column (CH.sub.2Cl.sub.2/EtOAc=50/1 to 25/1) to obtain PW0165 (793 mg, 91%) as a white solid. .sup.1NMR (300 MHz, Methanol-d.sub.4) δ 4.54-4.39 (m, 2H), 4.23 (d, J=6.7 Hz, 1H), 3.69 (dd, J=8.7, 5.4 Hz, 1H), 3.33 (q, J=1.6 Hz, 1H), 2.96 (tdd, J=10.4, 6.3, 3.9 Hz, 2H), 2.34 (dt, J=8.6, 6.5 Hz, 2H), 2.21-2.01 (m, 3H), 1.98-1.59 (m, 7H), 1.46 (s, 9H), 1.06-0.88 (m, 12H). .sup.13C NMR (75 MHz, Methanol-d.sub.4) δ 175.81, 174.44, 173.34, 172.58, 171.94, 80.40, 60.11, 58.28, 52.46, 51.52, 47.33, 47.05, 46.76, 46.67, 40.82, 31.19, 30.70, 30.60, 26.98, 26.86, 25.67, 24.65, 22.01, 20.72, 18.38, 17.04.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-2-((S)-1-benzoylpyrrolidine-2-carboxamido)-4-methylpentanamido)-5-oxopentanoate (PW0169)
[0102] PW0165 (102 mg, 0.2 mmol) was dissolved in 5 mL DCM and the solution was cooled to 0° C. with ice bath. Then Et.sub.3N (65 mg, 0.5 mmol) and benzoyl chloride (42 mg, 0.3 mmol) were added. The mixture was stirred at room temperature for overnight. The solution was diluted with 20 mL of DCM and washed with 1N Na.sub.2SO.sub.4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na2SO4, the solution was concentrated and purified with silica gel column (CHCl.sub.3/CH.sub.3OH=50/1 to 20/1) to obtain PW0169 (104 mg, 85%) as a white solid. .sup.1H NMR (300 MHz, Methanol-d.sub.4) δ 7.62 (dd, J=7.6, 2.0 Hz, 2H), 7.56-7.38 (m, 5H), 4.60 (dd, J=8.1, 5.8 Hz, 1H), 4.44 (dt, J=9.8, 5.2 Hz, 3H), 4.22 (t, J=6.2 Hz, 2H), 3.79-3.65 (m, 2H), 3.58 (dd, J=6.8, 4.7 Hz, 1H), 2.44-2.27 (m, 4H), 2.15-1.80 (m, 9H), 1.67 (dt, J=8.5, 5.9 Hz, 2H), 1.45 (s, 9H), 1.42-1.37 (m, 4H), 1.01-0.95 (m, 12H), 0.85 (dd, J=12.7, 6.4 Hz, 2H). .sup.13C NMR (75 MHz, Methanol-d.sub.4) δ 173.53, 172.48, 172.07, 130.14, 128.05, 126.92, 126.40, 80.35, 60.72, 58.39, 52.66, 52.24, 50.31, 47.31, 47.03, 46.74, 40.18, 31.18, 30.53, 29.59, 26.96, 26.83, 25.01, 24.51, 22.05, 20.64, 18.37, 17.33, 17.02, 15.89.
tert-Butyl (S)-4-((S)-2-((S)-1-(((3R,5R,7R)-adamantan-1-yl)carbamoyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0173)
[0103] PW0165 (51 mg, 0.1 mmol) was dissolved in 5 mL DCM and the solution was cooled to 0° C. with ice bath. Then Et.sub.3N (33 mg, 0.25 mmol) and 1-adamantyl isocyanate (18 mg, 0.1 mmol) were added. The mixture was stirred at room temperature overnight. The solution was diluted with 20 mL of DCM and washed with 1N NaHSO.sub.4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CHCl.sub.3/CH.sub.3OH=50/1 to 20/1) to obtain PW0173 (57 mg, 84%) as a white solid. .sup.1H NMR (300 MHz, Methanol-d.sub.4) δ 4.44 (dd, J=9.2, 5.4 Hz, 1H), 4.37-4.25 (m, 2H), 4.22 (d, J=6.6 Hz, 1H), 3.51 (d, J=7.1 Hz, 1H), 2.34 (td, J=7.6, 7.1, 2.5 Hz, 2H), 2.25-2.12 (m, 3H), 2.11-1.93 (m, 13H), 1.78-1.60 (m, 9H), 1.46 (s, 9H), 1.08-0.85 (m, 12H). .sup.13C NMR (75 MHz, Methanol-d.sub.4) δ 174.94, 173.76, 172.27, 80.38, 60.55, 58.63, 52.96, 52.56, 51.18, 46.29, 41.79, 40.07, 36.16, 31.26, 30.41, 29.66, 29.54, 26.95, 26.73, 24.60, 24.39, 22.01, 20.57, 18.34, 17.09.
tert-Butyl (S)-4-((S)-2-((S)-1-((3S,5S,7S)-adamantane-1-carbonyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0183)
[0104] PW0165 (51 mg, 0.1 mmol) and 1-adamantanecarboxylic acid (18 mg, 0.1 mmol) were dissolved in 5 mL DCM and the mixture solution was cooled to 0° C. with ice bath. HOBt (14 mg, 0.1 mmol), HBTU (76 mg, 0.2 mmol) and DIPEA (32 μL, 0.2 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl2/MeOH=50/1) to obtain PW0183 (49 mg, 73%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.84 (d, J=6.2 Hz, 1H), 7.10 (d, J=9.0 Hz, 1H), 6.93-6.74 (m, 2H), 5.33 (s, 1H), 4.62 (s, 1H), 4.43 (dd, J=9.0, 5.3 Hz, 1H), 4.28 (q, J=6.3, 5.7 Hz, 1H), 4.16-4.04 (m, 1H), 3.97-3.65 (m, 3H), 3.18 (dd, J=7.5, 4.4 Hz, 1H), 2.45 (d, J=3.6 Hz, 3H), 2.24-2.00 (m, 11H), 1.83-1.66 (m, 7H), 1.52-1.37 (m, 15H), 1.11-0.80 (m, 12H)..sup.13C NMR (75 MHz, Chloroform-d) δ 173.95, 171.68, 77.19, 62.79, 58.59, 55.18, 55.02, 54.04, 48.78, 42.18, 40.21, 38.18, 36.41, 32.34, 29.08, 28.13, 28.09, 26.56, 26.31, 26.19, 25.19, 23.19, 21.42, 19.47, 18.65, 17.48, 17.30.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-2-((S)-1-decanoylpyrrolidine-2-carboxamido)-4-methylpentanamido)-5-oxopentanoate (PW0184)
[0105] PW0165 (51 mg, 0.1 mmol) and capric acid (17 mg, 0.1 mmol) were dissolved in 5 mL of DCM and the mixture solution was cooled to 0° C. with ice bath. HOBt (14 mg, 0.1 mmol), HBTU (76 mg, 0.2 mmol) and DIPEA (32 μL, 0.2 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1) to obtain PW0184 (48 mg, 72%) as a white solid. .sup.1H NMR (300 MHz, DMSO-d6) δ 8.33-7.77 (m, 2H), 7.65-7.24 (m, 2H), 7.06 (s, 1H), 4.44-4.04 (m, 4H), 3.69-3.40 (m, 3H), 2.32-1.60 (m, 12H), 1.39 (s, 12H), 1.26 (dd, J=11.1, 4.5 Hz, 14H), 0.84 (m, 14H)..sup.13C NMR (75 MHz, DMSO-d.sub.6) δ 173.06, 172.57, 172.40, 172.14, 171.21, 80.07, 60.13, 59.70, 57.82, 53.94 , 52.36, 51.80, 42.19, 40.71, 40.40, 39.57, 34.25, 33.88, 32.15, 31.74, 31.67, 30.87, 29.66, 29.46, 29.39, 29.28, 29.20, 29.12, 28.20, 27.67, 27.38, 24.84, 24.78, 24.71, 24.63, 23.50, 23.44, 22.75, 22.54, 21.89, 19.68, 18.51, 18.11, 17.19, 14.39, 12.81.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-2-((S)-1-(cyclohexanecarbonyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-oxopentanoate (PW0185)
[0106] PW0165 (51 mg, 0.1 mmol) and cyclohexanecarboxylic acid (13 mg, 0.1 mmol) were dissolved in 5 mL of DCM and the mixture solution was cooled to 0° C. with ice bath. HOBt (14 mg, 0.1 mmol), HBTU (76 mg, 0.2 mmol) and DIPEA (32 μL, 0.2 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1) to obtain PW0185 (51 mg, 82%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 8.00 (s, 1H), 7.39 (s, 1H), 7.18 (s, 1H), 6.89 (s, 1H), 5.36 (s, 1H), 4.59 (s, 1H), 4.40 (s, 1H), 4.26 (d, J=8.6 Hz, 1H), 4.07 (s, 1H), 3.64 (d, J=33.5 Hz, 4H), 3.14 (d, J=7.9 Hz, 2H), 2.58-2.36 (m, 4H), 2.15 (t, J=31.3 Hz, 8H), 1.74 (s, 7H), 1.50 (d, J=8.4 Hz, 14H), 1.06-0.87 (m, 12H). .sup.13C NMR (75 MHz, Chloroform-d) δ 174.08, 173.65, 173.09, 77.19, 60.34, 58.72, 55.10, 54.36, 54.17, 47.41, 42.78, 42.50, 40.04, 32.26, 29.27, 29.09, 28.43, 28.07, 27.53, 25.97, 25.67, 25.60, 25.49, 25.14, 24.95, 23.13, 21.17, 19.43, 18.62, 17.47, 17.33, 12.18.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-2-((S)-1-(4-fluorobenzoyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-oxopentanoate (PW0192)
[0107] PW0192 (52 mg, 83%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 8.20-7.81 (m, 2H), 7.69-7.47 (m, 2H), 7.45-7.22 (m, 3H), 7.20-6.90 (m, 2H), 4.52-3.99 (m, 4H), 3.69-3.46 (m, 2H), 2.32-2.07 (m, 3H), 2.00-1.65 (m, 6H), 1.56-1.21 (m, 11H), 1.04-0.51 (m, 12H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) δ 173.04, 172.17, 171.97, 171.13, 130.36, 130.24, 129.70, 115.69, 115.42, 80.09, 61.68, 60.59, 57.71, 51.68, 50.36, 40.69, 40.40, 39.84, 39.28, 31.61, 31.06, 30.97, 29.89, 28.19, 27.59, 25.34, 24.66, 24.30, 23.46, 22.74, 22.09, 21.68, 19.68, 18.12.
tert-Butyl (S)-5-(((S)-1-amino-3-methyl-1-oxobutan-2-yl)amino)-4-((S)-2-((S)-1-((4-fluorophenyl)sulfonyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-oxopentanoate (PW0197)
[0108] PW0197 (47 mg, 70%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 8.17 (d, J=8.2 Hz, 1H), 8.01 (d, J=7.9 Hz, 1H), 7.93 (dd, J=8.7, 5.2 Hz, 2H), 7.49 (dt,J=17.1, 8.7 Hz, 3H), 7.38 (s, 1H), 7.05 (s, 1H), 4.30 (t, J=8.4 Hz, 2H), 4.12 (q, J=7.6 Hz, 2H), 3.49-3.36 (m, 3H), 3.17 (d, J=9.0 Hz, 2H), 2.21 (q, J=8.4, 7.7 Hz, 2H), 1.92 (td, J=13.6, 6.5 Hz, 4H), 1.79-1.68 (m, 4H), 1.58-1.45 (m, 4H), 1.37 (s, 9H), 0.85 (dt, J=18.9, 7.0 Hz, 12H). .sup.13C NMR (75 MHz, DMSO-d.sub.6) δ 173.04, 172.34, 172.16, 171.36, 171.09, 130.92, 130.80, 117.06, 116.76, 80.08, 61.35, 57.61, 52.23, 51.32, 49.49, 41.14, 31.68, 31.22, 31.05, 28.19, 27.54, 24.62, 24.55, 23.53, 22.08, 19.68, 18.12.
##STR00016##
tert-Butyl (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0157)
[0109] Fmoc-Glu(O.sup.tBu)-OH (1.3 g, 3 mmol) and H-Val-OMe⋅HCl (504 mg, 3 mmol) were dissolved in 20 mL of DCM and the mixture solution was cooled to 0° C. with ice bath. HOBt (405 mg, 3 mmol), HBTU (2.3 g, 6 mmol) and DIPEA (2 mL, 12 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1) to obtain PW0157 (1.53 g, 89%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.78 (d, J=7.4 Hz, 2H), 7.61 (d, J=7.4 Hz, 2H), 7.46-7.38 (m, 2H), 7.33 (td, J=7.4, 1.2 Hz, 2H), 6.98 (d, J=8.5 Hz, 1H), 5.77 (d, J=7.7 Hz, 1H), 4.53 (dd, J=8.6, 4.8 Hz, 1H), 4.45-4.37 (m, 2H), 4.25 (d, J=7.1 Hz, 2H), 3.75 (s, 3H), 2.46 (t, J=6.6 Hz, 2H), 2.29-1.94 (m, 3H), 1.49 (s, 9H), 0.96 (dd, J=8.9, 6.9 Hz, 6H).
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0164)
[0110] To a solution of DEA (10 mL) and MeCN (10 mL) was added PW0157 (1.08 g, 2 mmol) and the solution was stirred at room temperature for 1 h. After the reaction completed, the solution was concentrated. The crude residue was dissolved in 20 mL of dry DCM and added Z-Pro-Leu-OH (724, 2 mmol). The mixture solution was cooled to 0° C. with ice bath. HOBt (270 mg, 2 mmol), HBTU (1.5 g, 4 mmol) and DIPEA (1.3 mL, 8 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1 to 20/1) to obtain PW0164 (1.24 g, 94%) as a light yellow solid. .sup.1NMR (300 MHz, Chloroform-d) δ 7.36 (s, 6H), 7.10 (d, J=8.6 Hz, 1H), 6.90-6.78 (m, 1H), 5.17 (s, 2H), 4.54-4.31 (m, 4H), 3.73 (s, 3H), 3.54 (d, J=8.3 Hz, 2H), 2.39 (dt, J=11.0, 7.3 Hz, 2H), 2.20 (pd, J=6.9, 5.3 Hz, 3H), 2.04-1.89 (m, 3H), 1.44 (s, 9H), 0.92 (ddd, J=13.8, 8.3, 5.4 Hz, 12H). .sup.13C NMR (75 MHz, Chloroform-d) δ 173.07, 171.94, 171.02, 136.23, 128.53, 128.17, 127.86, 80.86, 67.54, 60.93, 57.51, 52.38, 52.03, 47.15, 40.45, 31.97, 30.79, 28.86, 28.07, 27.26, 25.02, 24.69, 22.94, 21.58, 18.99, 17.81. HRMS (ESI) calcd for C34H52N4O9 661.3813 (M+H).sup.+, found 661.3804.
tert-Butyl (S)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino) -4-((S)-4-methyl-2-((S)-pyrrolidine-2-carboxamido)pentanamido)-5-oxopentanoate (PW0539)
[0111] To a solution of PW0164 (1.0 g) in 100 mL MeOH, 10% Pd/C (100 mg) was added. Under H.sub.2, the mixture was allowed to stir at room temperature for 12 hrs. The solution was filtered, and the filtrate was concentrated to get the crude product. The residue was purified by silica gel column (CH.sub.2Cl.sub.2/MeOH=20/1 to 15/1) to obtain PW0539 (733 mg, 92%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 8.00 (d, J=8.3 Hz, 1H), 7.32-7.24 (m, 1H), 7.19 (d, J=8.5 Hz, 1H), 4.55-4.36 (m, 3H), 3.82 (dd, J=9.1, 5.3 Hz, 1H), 3.72 (s, 3H), 3.55-3.27 (m, 1H), 3.06-2.91 (m, 2H), 2.82 (s, 2H), 2.39 (q, J=6.9 Hz, 2H), 2.21-2.03 (m, 3H), 1.90 (tt, J=12.7, 6.4 Hz, 2H), 1.77-1.54 (m, 6H), 1.44 (s, 9H), 1.25 (t, J=7.1 Hz, 1H), 1.10 (t, J=7.2 Hz, 1H), 0.95-0.88 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 175.26, 173.22, 172.33, 171.98, 171.12, 80.98, 60.38, 57.53, 52.48, 52.04, 51.47, 47.22, 40.84, 31.73, 30.73, 30.67, 28.05, 27.46, 26.05, 24.93, 22.95, 21.78, 18.97, 17.75.
tert-Butyl (S)-4-((S)-2-((S)-1-((3S,5S,7S)-adamantane-1-carbonyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0564)
[0112] PW0564 (46 mg, 67%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.28-7.22 (m, 1H), 7.13 (d, J=8.6 Hz, 1H), 6.74 (d, J=7.0 Hz, 1H), 4.59 (q, J=3.8 Hz, 1H), 4.43 (td, J=8.3, 4.1 Hz, 2H), 4.29 (td, J=6.8, 3.7 Hz, 1H), 3.79 (ddd, J=18.8, 8.1, 3.4 Hz, 2H), 3.69 (s, 3H), 3.44-3.20 (m, 1H), 2.45-2.31 (m, 2H), 2.15 (dt, J=13.2, 6.7 Hz, 3H), 2.03-1.91 (m, 15H), 1.74-1.64 (m, 9H), 1.41 (s, 9H), 1.23 (s, 1H), 1.07 (t, J=7.1 Hz, 1H), 0.88 (dd, J=14.8, 6.5 Hz, 14H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 177.64, 172.75, 172.30, 172.02, 171.86, 171.10, 80.64, 62.28, 57.56, 52.60, 52.41, 51.91, 48.55, 42.01, 40.46, 38.14, 38.12, 36.58, 36.46, 32.01, 30.70, 28.30, 28.18, 28.03, 27.32, 24.94, 23.15, 21.50, 18.99, 17.94.
tert-Butyl (S)-4-((S)-2-((S)-1-acetylpyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0565)
[0113] PW0565 (45 mg, 78%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.31-7.23 (m, 2H), 7.17 (d, J=8.5 Hz, 1H), 4.59-4.41 (m, 3H), 4.32 (ddd, J=9.3, 7.0, 4.7 Hz, 1H), 3.71 (s, 3H), 3.57 (q, J=5.5, 3.5 Hz, 1H), 3.46 (q, J=9.4, 8.6 Hz, 1H), 2.47-2.16 (m, 6H), 2.10 (s, 3H), 2.05-1.92 (m, 4H), 1.71 (d, J=12.9 Hz, 1H), 1.59 (dt, J=14.4, 7.6 Hz, 2H), 1.44 (s, 9H), 0.95-0.85 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.18, 172.04, 171.92, 171.70, 171.10, 80.85, 59.86, 57.49, 52.60, 52.48, 51.99, 48.29, 40.42, 31.87, 30.76, 28.07, 27.83, 27.38, 25.07, 24.97, 22.94, 22.43, 21.60, 18.98, 17.79.
(9H-Fluoren-9-yl)methyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S) -1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0576)
[0114] PW0576 (55 mg, 73%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.78 (d, J=7.6 Hz, 2H), 7.60 (d, J=7.4 Hz, 2H), 7.42 (t, J=7.3 Hz, 2H), 7.33 (t, J=7.4 Hz, 2H), 7.28 (s, 1H), 7.08 (d, J=8.6 Hz, 1H), 6.85 (d, J=7.4 Hz, 1H), 4.51-4.22 (m, 7H), 3.73 (s, 3H), 3.60-3.41 (m, 2H), 3.34-3.19 (m, 1H), 2.40 (dq, J=16.0, 7.0, 5.8 Hz, 1H), 2.27-2.14 (m, 3H), 1.96 (s, 3H), 1.78-1.57 (m, 2H), 1.43 (s, 9H), 0.91 (ddd, J=13.9, 6.5, 4.0 Hz, 13H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.15, 171.90, 170.98, 156.35, 143.73, 141.33, 141.26, 127.78, 127.63, 127.08, 125.17, 124.95, 120.00, 119.86, 80.88, 67.81, 60.82, 57.48, 52.62, 52.31, 52.02, 47.20, 47.05, 41.75, 41.28, 40.49, 31.96, 30.78, 28.05, 27.29, 25.02, 24.74, 23.45, 22.94, 21.55, 18.99, 17.78, 14.19, 13.81.
##STR00017##
Allyl (((9H-fluoren-9-yl)methoxy)carbonyl)-L-valinate (PW0529)
[0115] To a solution of Fmoc-Val-OH (1.7 g, 5 mmol) in 5 mL dry DMF, K.sub.2CO.sub.3 (1.0 g, 7.5 mmol), allyl bromide (1.2 g, 10 mmol) and TBAB (161 mg, 0.5 mmol) were added. The mixture was stirred at room temperature for 16 h. After the reaction completed (detected by TLC), the mixture was diluted with 20 mL of water. Then the mixture was extracted with EtOAc (3×20 mL). The combined organic layer was washed with 1N NaHSO.sub.4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1 to 20/1) to obtain PW0529 (1.5 g, 76%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.79 (dd, J=7.4, 1.1 Hz, 2H), 7.63 (d, J=7.5 Hz, 2H), 7.43 (tt, J=7.7, 1.4 Hz, 2H), 7.34 (tt, J=7.4, 1.1 Hz, 2H), 6.00-5.88 (m, 1H), 5.45-5.21 (m, 4H), 4.75-4.57 (m, 3H), 4.43 (d, J=7.1 Hz, 1H), 4.37 (dd, J=9.1, 4.8 Hz, 1H), 4.26 (t, J=7.0 Hz, 1H), 2.23 (td, J=6.9, 4.6 Hz, 1H), 1.03-0.93 (m, 6H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 171.76, 156.21, 143.78, 141.31, 131.57, 127.69, 127.05, 125.08, 119.97, 119.02, 67.05, 65.86, 59.04, 47.23, 31.36, 18.99, 17.56.
tert-Butyl (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((S)-1-(allyloxy)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0530)
[0116] To a solution of DEA (10 mL) and MeCN (10 mL) was added PW0529 (1.5 g, 4 mmol) and the solution was stirred at room temperature for 1 h. After the reaction completed, the solution was concentrated. The crude residue was dissolved in 20 mL of dry DCM and added Fmoc- Glu(O.sup.tBu)-OH (1.7, 4 mmol). The mixture solution was cooled to 0° C. with ice bath. HOBt (540 mg, 4 mmol), HBTU (2.3 g, 6 mmol) and DIPEA (2.6 mL, 16 mmol) were added to the solution at 0° C. Then removed the ice bath, and the mixture solution was stirred at room temperature overnight. After the reaction completed (detected by TLC), the mixture was washed with 1N NaHSO4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1 to 20/1) to obtain PW0530 (1.5 g, 68%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.78 (d, J=7.5 Hz, 2H), 7.61 (d, J=7.4 Hz, 2H), 7.42 (t, J=7.4 Hz, 2H), 7.33 (td, J=7.4, 1.2 Hz, 2H), 7.00 (d, J=8.6 Hz, 1H), 5.92 (tt, J=10.9, 5.4 Hz, 1H), 5.77 (d, J=7.7 Hz, 1H), 5.31 (dd, J=25.1, 13.9 Hz, 2H), 4.65 (s, 2H), 4.55 (dd, J=8.6, 4.7 Hz, 1H), 4.40 (dd, J=7.2, 2.1 Hz, 2H), 4.28 (dt, J=28.2, 6.7 Hz, 2H), 2.64-2.37 (m, 2H), 2.25 (dd, J=12.4, 6.4 Hz, 1H), 2.16-2.07 (m, 1H), 1.98 (dd, J=14.3, 7.2 Hz, 1H), 1.49 (s, 9H), 0.97 (dd, J=9.8, 6.9 Hz, 6H).
Benzyl (S)-2-(((S)-1-(((S)-1-(((S)-1-(allyloxy)-3-methyl-1-oxobutan -2-yl)amino)-5-(tert-butoxy)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0531)
[0117] PW0531 (1.46 g, 88%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.36 (s, 5H), 7.12 (d, J=8.6 Hz, 1H), 6.83 (d, J=7.4 Hz, 1H), 5.92 (ddt, J=16.5, 11.0, 5.7 Hz, 1H), 5.34 (dq, J=17.2, 1.5 Hz, 1H), 5.28-5.21 (m, 1H), 5.17 (s, 2H), 4.63 (td, J=5.1, 4.4, 1.5 Hz, 2H), 4.53-4.31 (m, 4H), 3.53 (dd, J=12.6, 6.7 Hz, 2H), 2.50-2.34 (m, 2H), 2.30-2.12 (m, 3H), 2.05-1.83 (m, 4H), 1.44 (s, 10H), 0.98-0.82 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.09, 171.94, 171.11, 171.00, 166.59, 156.32, 136.25, 131.76, 128.53, 128.17, 127.86, 118.67, 80.84, 67.52, 65.66, 60.88, 57.49, 52.63, 52.32, 47.13, 40.50, 32.00, 30.82, 28.79, 28.07, 27.32, 25.00, 24.69, 22.95, 21.59, 19.03, 17.73.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-3-methyl-1-morpholino-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl -1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0536)
[0118] To a solution of PW0531 (69 mg, 0.1 mmol) in CH.sub.2Cl.sub.2 (5 mL) at 0° C. was added Pd(PPh3)4 (12 mg, 0.01 mmol) and DEA (0.1 mL, 1 mmol). The reaction was stirred at room temperature for 10 h. After evaporation in vacuo, the residue was dissolved into CH.sub.2Cl.sub.2 (5 mL). The mixture solution was cooled to 0° C. with ice bath. Then morpholine (0.015 mL, 0.2 mmol) HOBt (14 mg, 40.1 mmol), HBTU (57 mg, 0.15 mmol) and DIPEA (0.07 mL, 0.4 mmol) were added to the solution at 0° C. After the removal of the ice bath, the mixture solution was stirred at room temperature overnight. The mixture was washed with 1N NaHSO.sub.4, saturated NaHCO.sub.3 and Brine. After drying over anhydrous Na.sub.2SO.sub.4, the solution was concentrated and purified with silica gel column (CH.sub.2Cl.sub.2/MeOH=50/1 to 20/1) to obtain PW0536 (57 g, 80%) as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.34 (s, 5H), 7.27-7.17 (m, 1H), 7.12-6.64 (m, 1H), 5.16 (s, 2H), 4.67 (t, J=8.0 Hz, 1H), 4.49 (td, J=8.2, 5.0 Hz, 1H), 4.35 (q, J=7.6, 5.5 Hz, 2H), 3.87-3.48 (m, 10H), 3.35 (d, J=4.6 Hz, 1H), 2.98 (d, J=8.5 Hz, 2H), 2.49-1.82 (m, 10H), 1.77-1.49 (m, 3H), 1.42 (s, 10H), 1.01-0.77 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 172.61, 172.28, 172.01, 170.84, 169.76, 166.61, 156.14, 136.29, 128.50, 128.15, 127.81, 80.72, 67.48, 66.83, 66.72, 65.98, 60.90, 53.50, 52.76, 52.42, 50.61, 48.79, 47.12, 46.30, 42.42, 40.37, 38.57, 31.92, 31.00, 28.99, 28.06, 27.32, 24.98, 24.66, 22.90, 21.67, 19.64, 17.64.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-1-(diethylamino)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0540)
[0119] PW0540 (46 mg, 65%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.31 (m, 5H), 7.05 (m, 2H), 5.16 (s, 2H), 4.81-4.19 (m, 4H), 3.48 (dq, J=32.6, 6.2, 5.5 Hz, 4H), 3.19 (dd, J=13.6, 7.0 Hz, 1H), 2.40-1.85 (m, 9H), 1.57 (d, J=31.5 Hz, 2H), 1.42 (d, J=2.0 Hz, 9H), 1.21 (t, J=7.1 Hz, 3H), 1.10 (t, J=7.0 Hz, 3H), 1.02-0.76 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 172.44, 171.90, 170.67, 170.51, 156.00, 136.42, 131.99, 128.54, 128.48, 128.45, 128.06, 127.80, 80.60, 67.36, 60.63, 53.87, 52.61, 47.08, 42.07, 40.68, 40.40, 31.71, 29.66, 28.92, 28.05, 24.88, 22.89, 21.80, 19.58, 19.49, 17.80, 14.62, 14.57, 12.87, 11.74.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-3-methyl-1-(methylamino)-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0541)
[0120] PW0541 (56 mg, 86%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.87 (d, J=6.2 Hz, 1H), 7.38 (s, 5H), 7.02 (d, J=5.2 Hz, 1H), 6.91 (d, J=9.3 Hz, 1H), 6.74 (s, 1H), 5.20 (s, 2H), 4.49-4.12 (m, 4H), 3.57 (d, J=7.2 Hz, 2H), 2.80 (d, J=4.7 Hz, 3H), 2.53-2.31 (m, 3H), 2.25-2.05 (m, 4H), 2.02-1.90 (m, 2H), 1.67 (d, J=10.8 Hz, 4H), 1.45 (s, 9H), 0.92 (dd, J=17.1, 6.0 Hz, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.44, 173.18, 172.69, 171.55, 171.37, 156.59, 136.07, 128.61, 128.35, 127.82, 81.15, 67.73, 60.93, 58.68, 54.77, 53.71, 47.18, 40.22, 32.32, 29.15, 28.61, 28.08, 26.23, 25.10, 24.72, 22.96, 21.41, 19.44, 17.28.
Benzyl (S)-2-(((S)-1-(((S)-1-(((S)-1-(benzylamino)-3-methyl-1-oxobutan-2-yl)amino)-5-(tert-butoxy)-1,5-dioxopentan-2-yl)amino)-4-methyl -1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0542)
[0121] PW0542 (57 mg, 75%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.87 (d, J=6.4 Hz, 1H), 7.37 (s, 5H), 7.28 (s, 5H), 7.22-7.16 (m, 2H), 7.09 (d, J=8.7 Hz, 1H), 6.99 (d, J=5.7 Hz, 1H), 5.18 (s, 2H), 4.59-4.27 (m, 6H), 3.52 (t, J=7.2 Hz, 2H), 2.40 (s, 4H), 2.20-1.93 (m, 6H), 1.60 (s, 2H), 1.44 (s, 9H), 0.92 (dd, J=17.3, 6.1 Hz, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.21, 172.90, 172.53, 172.09, 171.49, 171.02, 156.41, 138.66, 136.14, 128.57, 128.26, 127.83, 127.64, 126.84, 81.01, 67.65, 60.92, 58.65, 54.48, 53.40, 47.12, 43.26, 40.35, 39.84, 32.24, 29.59, 28.75, 28.08, 26.56, 25.01, 24.69, 22.89, 21.56, 19.53, 19.46, 17.52.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-3-methyl-1-oxo-1-(pyrrolidin-1-yl)butan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0543)
[0122] PW0543 (53 mg, 76%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.34 (s, 5H), 7.21 (d, J=8.6 Hz, 1H), 7.15 (t, J=4.3 Hz, 1H), 6.98 (s, 1H), 5.16 (s, 2H), 4.44 (dt, J=41.0, 6.4 Hz, 4H), 3.73-3.38 (m, 7H), 2.29 (d, J=7.9 Hz, 2H), 2.08-1.83 (m, 10H), 1.41 (s, 9H), 0.89 (dt, J=11.9, 5.0 Hz, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 172.54, 172.28, 171.99, 170.83, 169.72, 156.04, 136.38, 128.46, 128.06, 127.80, 80.63, 67.37, 60.73, 56.03, 52.67, 49.44, 47.08, 46.72, 45.84, 40.12, 31.09, 29.65, 29.01, 28.05, 27.60, 26.01, 25.07, 24.91, 24.16, 22.89, 19.45, 17.87.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-1-(3-hydroxyazetidin-1-yl)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0544)
[0123] PW0544 (46 mg, 66%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.51 (s, 1H), 7.35 (s, 5H), 7.15 (d, J=8.7Hz, 1H), 6.87 (s, 1H), 5.17 (s, 2H), 4.64-3.99 (m, 9H), 3.55 (q, J=8.1, 6.8 Hz, 2H), 2.45-1.81 (m, 10H), 1.75-1.50 (m, 3H), 1.43 (s, 9H), 0.91 (d, J=6.5 Hz, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 172.75, 172.46, 171.26, 170.74, 156.43, 136.44, 128.53, 128.19, 127.80, 80.92, 67.56, 61.25, 60.37, 58.26, 57.61, 54.34, 53.68, 52.92, 47.20, 40.29, 31.94, 28.07, 27.26, 25.04, 24.67, 22.91, 21.63, 19.19, 18.14.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-1-(cyclohexylamino)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0545)
[0124] PW0545 (54 mg, 74%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.85 (d, J=6.4 Hz, 1H), 7.34 (d, J=14.5 Hz, 5H), 7.04 (d, J=5.4 Hz, 1H), 7.00 (s, 1H), 6.64 (s, 1H), 5.18 (s, 2H), 4.49-4.24 (m, 4H), 3.72 (s, 1H), 3.55 (q, J=7.4, 6.5 Hz, 2H), 2.38 (d, J=7.1 Hz, 2H), 2.13 (dq, J=16.0, 8.6, 7.1 Hz, 6H), 1.90 (dt, J=24.2, 9.1 Hz, 6H), 1.77-1.50 (m, 8H), 1.43 (s, 9H), 1.34 (d, J=3.0 Hz, 2H), 1.24-1.13 (m, 3H), 0.90 (dd, J=15.6, 7.7 Hz, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.12, 172.54, 171.32, 169.82, 156.41, 136.16, 128.55, 128.28, 127.84, 80.93, 67.65, 61.01, 58.64, 54.85, 54.35, 53.38, 48.34, 47.16, 40.41, 39.93, 33.39, 32.90, 32.70, 32.25, 29.57, 28.88, 28.07, 26.64, 25.57, 25.04, 25.00, 24.72, 22.89, 21.58, 19.40, 17.45.
Benzyl (S)-2-(((S)-1-(((S)-1-(((S)-1-(((3R,5R,7R)-adamantan-1-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-(tert-butoxy)-1,5-dioxopentan -2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0547)
[0125] PW0547 (55 mg, 71%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.69 (d, J=5.3 Hz, 1H), 7.37 (s, 5H), 6.95 (d, J=13.6 Hz, 2H), 6.08 (s, 1H), 5.18 (s, 2H), 4.60-4.19 (m, 4H), 3.54 (s, 2H), 2.38 (s, 3H), 2.13 (d, J=7.6 Hz, 3H), 2.01 (d, J=13.1 Hz, 13H), 1.88 (s, 2H), 1.66 (s, 9H), 1.44 (s, 9H), 0.93 (d, J=6.6 Hz, 8H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 173.09, 172.35, 171.93, 171.14, 169.84, 169.78, 156.38, 136.19, 132.14, 132.01, 128.55, 128.51, 128.27, 127.84, 80.88, 80.67, 67.62, 60.93, 59.13, 53.97, 53.16, 51.99, 47.15, 41.47, 41.26, 40.51, 36.40, 32.22, 29.70, 29.47, 29.42, 28.79, 28.08, 26.77, 25.03, 24.72, 23.09, 22.91, 21.59, 19.46, 19.37, 18.03, 17.58.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-3-methyl-1-(4-methylpiperazin-1-yl)-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino) -4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0559)
[0126] PW0559 (44 mg, 60%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.34 (s, 5H), 7.18 (d, J=8.3 Hz, 1H), 6.91 (s, 1H), 5.16 (s, 2H), 4.74 (t, J=7.7 Hz, 1H), 4.50 (q, J=8.1, 5.5 Hz, 1H), 4.36 (q, J=8.2, 5.5 Hz, 2H), 3.74-3.45 (m, 6H), 2.39 (t, J=9.6 Hz, 5H), 2.30 (s, 4H), 2.18-1.87 (m, 6H), 1.57 (d, J=37.3 Hz, 3H), 1.42 (s, 9H), 1.06-0.81 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 172.57, 172.14, 171.91, 170.75, 169.51, 156.12, 136.34, 128.48, 128.10, 127.81, 80.68, 67.42, 60.78, 55.14, 54.68, 53.45, 52.71, 52.31, 47.09, 45.91, 45.68, 41.99, 40.56, 31.87, 31.22, 29.66, 28.91, 28.07, 27.49, 24.94, 24.66, 22.89, 21.75, 19.76, 19.69, 17.45.
Benzyl (S)-2-(((S)-1-(((S)-5-(tert-butoxy)-1-(((S)-1-((2-hydroxyethyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)pyrrolidine-1-carboxylate (PW0561)
[0127] PW0561 (22 mg, 32%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ .07 (d, J=5.7 Hz, 1H), 7.38 (s, 5H), 7.17 (d, J=4.4 Hz, 1H), 7.02-6.88 (m, 2H), 5.20 (s, 2H), 4.44 (ddd, J=32.9, 8.5, 4.6 Hz, 2H), 4.17 (dd, J=35.1, 5.7 Hz, 2H), 3.68 (d, J=11.4 Hz, 2H), 3.56 (tt, J=7.2, 3.9 Hz, 2H), 3.35 (d, J=5.8 Hz, 1H), 2.50-2.34 (m, 2H), 2.14 (dq, J=8.0, 5.0 Hz, 3H), 2.05-1.87 (m, 3H), 1.65 (dd, J=9.4, 4.4 Hz, 2H), 1.45 (s, 9H), 0.97-0.81 (m, 12H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 174.04, 173.55, 173.47, 172.76, 171.43, 171.36, 156.71, 135.98, 128.63, 128.61, 128.41, 127.83, 81.32, 67.83, 61.73, 60.97, 58.40, 55.51, 54.31, 47.21, 42.84, 40.25, 32.42, 29.68, 28.94, 28.56, 28.09, 25.93, 25.07, 24.73, 22.82, 21.55, 19.54, 19.49, 17.34.
tert-Butyl (4S)-4-((2S)-2-((2S)-1-((1R,3R)-adamantane-1-carbonyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0564)
[0128] PW0564 (46 mg, 67%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.28-7.22 (m, 1H), 7.13 (d, J=8.6 Hz, 1H), 6.74 (d, J=7.0 Hz, 1H), 4.59 (q, J=3.8 Hz, 1H), 4.43 (td, J=8.3, 4.1 Hz, 2H), 4.29 (td, J=6.8, 3.7 Hz, 1H), 3.79 (ddd, J=18.8, 8.1, 3.4 Hz, 2H), 3.69 (s, 3H), 3.44-3.20 (m, 1H), 2.45-2.31 (m, 2H), 2.15 (dt, J=13.2, 6.7 Hz, 3H), 2.03-1.91 (m, 15H), 1.74-1.64 (m, 9H), 1.41 (s, 9H), 1.23 (s, 1H), 1.07 (t, J=7.1 Hz, 1H), 0.88 (dd, J=14.8, 6.5 Hz, 14H).
tert-Butyl (S)-4-((S)-2-((S)-1-acetylpyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0565)
[0129] PW0565 (45 mg, 83%) was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.31-7.23 (m, 2H), 7.17 (d, J=8.5 Hz, 1H), 4.59-4.41 (m, 3H), 4.32 (ddd, J=9.3, 7.0, 4.7 Hz, 1H), 3.71 (s, 3H), 3.57 (q, J=5.5, 3.5 Hz, 1H), 3.46 (q, J=9.4, 8.6 Hz, 1H), 2.47-2.16 (m, 6H), 2.10 (s, 3H), 2.05-1.92 (m, 4H), 1.71 (d, J=12.9 Hz, 1H), 1.59 (dt, J=14.4, 7.6 Hz, 2H), 1.44 (s, 9H), 0.95-0.85 (m, 12H).
tert-Butyl (4S)-4-((2S)-2-(1-(2-((9H-fluoren-9-yl)methoxy)-2-oxoacetyl)pyrrolidine-2-carboxamido)-4-methylpentanamido)-5-(((S)-1-methoxy-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (PW0576)
[0130] PW0576 was synthesized in a similar fashion as a white solid. .sup.1H NMR (300 MHz, Chloroform-d) δ 7.78 (d, J=7.6 Hz, 2H), 7.60 (d, J=7.4 Hz, 2H), 7.42 (t, J=7.3 Hz, 2H), 7.33 (t, J=7.4 Hz, 2H), 7.28 (s, 1H), 7.08 (d, J=8.6 Hz, 1H), 6.85 (d, J=7.4 Hz, 1H), 4.51-4.22 (m, 7H), 3.73 (s, 3H), 3.60-3.41 (m, 2H), 3.34-3.19 (m, 1H), 2.40 (dq, J=16.0, 7.0, 5.8 Hz, 1H), 2.27-2.14 (m, 3H), 1.96 (s, 3H), 1.78-1.57 (m, 2H), 1.43 (s, 9H), 0.91 (ddd, J=13.9, 6.5, 4.0 Hz, 13H).
[0131] It is to be understood that both the foregoing general description of the invention and the following detailed description are exemplary, and thus do not restrict the scope of the invention.
[0132] All publications mentioned herein are incorporated by reference to the extent they support the present invention.