CN101519427B - Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof - Google Patents

Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof Download PDF

Info

Publication number
CN101519427B
CN101519427B CN200810007611.3A CN200810007611A CN101519427B CN 101519427 B CN101519427 B CN 101519427B CN 200810007611 A CN200810007611 A CN 200810007611A CN 101519427 B CN101519427 B CN 101519427B
Authority
CN
China
Prior art keywords
formula
trp
dba
leu
phe
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN200810007611.3A
Other languages
Chinese (zh)
Other versions
CN101519427A (en
Inventor
刘克良
杨菁
梁远军
许笑宇
宫泽辉
董华进
孔令雷
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Pharmacology and Toxicology of AMMS
Original Assignee
Institute of Pharmacology and Toxicology of AMMS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Pharmacology and Toxicology of AMMS filed Critical Institute of Pharmacology and Toxicology of AMMS
Priority to CN200810007611.3A priority Critical patent/CN101519427B/en
Publication of CN101519427A publication Critical patent/CN101519427A/en
Application granted granted Critical
Publication of CN101519427B publication Critical patent/CN101519427B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention relates to a novel-structure compound with endothelin receptor antagonizing activity, a preparation method thereof, and drug composition comprising the compound and the application of the compound on treating or preventing the illness related with endothelin effects.

Description

There is the peptide derivative of endothelin receptor antagonizing activity and pharmaceutical composition thereof and purposes
Technical field
The present invention relates to the peptide derivative with endothelin receptor antagonizing activity, its preparation method, containing their pharmaceutical composition and their purposes in prevention or the treatment disease relevant with endothelin and prostate gland.
Background technology
Endothelin (Endothelin, be called for short ET) is secreted by blood vessel endothelium, and they are one of the strongest biotic factors of vasoconstriction effect that a class finds so far.There is the gene of three kinds of endothelin of encoding respectively in the mankind or other mammalian genes group, the big ET of formation, under the effect of Endothelin-converting Enzyme, changes ET-1, ET-2, ET-3 into.They are 21 peptides, containing two disulfide bridge bonds.ET-1 is not only expressed in non-vascular cell, and is the endothelin being uniquely present in vascular endothelial cell.ET-2 and ET-3 is mainly in brain, kidney and suprarenal gland and small intestine.The specific receptors of ET in target cell is combined, the corresponding biological effect of final generation.Three class ET acceptor, i.e. ETA, ETB and ETC are found so far.ETA is mainly distributed in the unstriated muscle of aorta, atrium, placenta, lung, the cerebrovascular and renal blood vessels; ETB is present in neurogliocyte in central nervous system, arteries and veins falls clump epithelial cell, lung, tire blood, renal glomerulus endothelium, ventricle, brain are medium, also has distribution at vascular smooth muscle; ETC is then mainly distributed in endotheliocyte.Three kinds of acceptors are different to the avidity of various ET.Endothelin and acceptor have physiopathology reaction widely in human body.Take part in as hypertension, congestive heart failure, myocardial ischemia, brain suffocate, suffer a shock, the process such as acute renal failure, pulmonary hypertension, vasospasm disease.Therefore ET acceptor is considered to the practicable novel targets for the treatment of cardiovascular and cerebrovascular diseases, and its receptor antagonist contributes to preventing/treating cardiovascular and cerebrovascular diseases.
Summary of the invention
The object of the invention is to find new peptide-like endothelin receptor antagonist.
The present inventor after deliberation, has now found that formula (I) or formula (II) peptide derivative
R-(CH 2) i-CO-(CH 2) j-AA 1-AA 2-AA 3-OH formula (I)
R-(CH 2) i-CO-(CH 2) j-AA 1(M)-AA 2-AA 3-OH formula (II)
Or its steric isomer or its pharmaceutical salts have good endothelin receptor antagonizing activity, therefore formula (I) or formula (II) peptide derivative or its steric isomer or its pharmaceutical salts can be used as medicine for preventing or treating the disease relevant with endothelin and prostate gland.
First aspect present invention relates to formula (I) peptide derivative or its steric isomer or its pharmaceutical salts,
R-(CH 2) i-CO-(CH 2) j-AA 1-AA 2-AA 3-OH formula (I)
Wherein, R is hexamethylene imine base or following group:
(structural formula 1) (structural formula 2) (structural formula 3) (structural formula 4)
In structural formula 1, n 1, n 2and n 3be integer 0,1,2 or 3 independently, and wherein n 3≠ 0,0≤n 1+ n 2≤ 4; X 1for NH, O, S or CH 2; X 2for N or CH; Work as n 1=n 2=1, n 3=2, X 1for CH 2, X 2during for atom N, R is abbreviated as ABO;
In structural formula 2, m 1, m 2, m 3and m 4be integer 0,1 or 2 independently, and wherein 0≤m 1+ m 2≤ 4,0≤m 3+ m 4≤ 4; y 1and y 3be CH independently 2, O, S or NH, y 2for N or CH; Work as m 1=m 2=1, m 3=m 4=2, y 1, y 3for O atom, y 2during for atom N, R is abbreviated as DAD; Work as m 1=m 2=1, m 3=m 4=2, y 3for CH 2, y 1for O atom, y 2during for atom N, R is abbreviated as CSO;
In structural formula 3, z is N or CH, k is N or Ck 1(k 1for H, CH 3or CH 2cH 3), t is H, CH 3, CH 2cH 3or CH 1(CH 3) 2; Or z is O or S, k is N or Ck 1(k 1for H, CH 3or CH 2cH 3), t and connecting key thereof do not exist; Work as z=N, when k=N, t=H, R is piperazinyl; Work as z=O, when k=N, t and connecting key thereof do not exist, R is morpholinyl;
In structural formula 4, a and b is integer 0,1,2 or 3 independently; P is atom N; Work as a=3, during b=3, R is abbreviated as DBA;
AA 1for L or D type Ser, Thr, Phe, Cys, Gln, Asn, Ala, Ile, Leu, Nle, Val, other L or D type aliphatics alpha-non-natural amino acid, as β-Ala, γ-aminobutyric acid or aminoisobutyric acid;
AA 2for L or D type Trp, Pro, aromatic amino acid, cis or trans Hyp or following cis or trans Hyp derivative (structural formula 5):
(structural formula 5)
In structural formula 5, R 1for linking group, can be-O-,-NH-,-NH-CO-,-O-CO-,-CH 2-,-NH-CO-O-,-O-CO-NH-,-NH-CO-NH-or-NH-SO 2-.R 2for aryl, virtue is amino, aryl-lower alkylamino, aryl-sulfo-, aryl-lower alkyl-sulfo-, aryloxy, aryl-lower alkoxyl group, aryl-lower alkyl, aryl-sulfinyl, heteroaryl, heteroaryl-oxo, heteroaryl-amino, heteroaryl-sulfo-, heteroaryl-short-chain alkyl, heteroaryl-sulfinyl, heterocyclic radical, heterocyclic radical-lower alkyloxy, heterocyclic radical-oxo, heterocyclic radical-amino, heterocyclic radical-lower alkylamino, heterocyclic radical-sulfo-, heterocyclic radical-short-chain alkyl-sulfo-, heterocyclic radical-short-chain alkyl, heterocyclic radical-sulfinyl, cycloalkyl, cycloalkyl-oxo, cycloalkyl-lower alkyloxy, cycloalkyl-amino, cycloalkyl-lower alkylamino, cycloalkyl-sulfo-, cycloalkyl-short-chain alkyl sulfo-, cycloalkyl-short-chain alkyl or cycloalkyl-sulfinyl.
AA 3for the non-natural aromatic amino acid of D type, tryptophane, phenylglycocoll or non-amino acid, the phenyl wherein in aromatic nucleus can be selected from halogen, nitro, urea groups, methoxyl group on 2,3,4 or 5, and the substituting group of carboxyl and short-chain alkyl is monosubstituted or two replace.
0≤i≤4; 0≤j≤4; Wherein i=0 and j=0 time, AA 2be only L or D type cis or trans Hyp derivative;
Work as AA 2be only L or D type cis or trans Hyp derivative time, AA 3can lack.
In the definition of formula (I), except as otherwise noted, expression lower refers to the straight or branched group with 1 to 7 carbon atom, preferentially has 1 to 4 carbon atom.The example of short-chain alkyl and lower alkyloxy is methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, amyl group, hexyl, heptyl, methoxyl group, oxyethyl group, propoxy-, n-butoxy, isobutoxy, sec-butoxy and tert.-butoxy.The example of lower alkenyl and lower alkinyl is vinyl, propenyl, butenyl, 2-methylpropenyl, ethynyl, proyl, butynyl, pentynyl, 2-methyl-pentinyl.Expressions cycloalkyl refers to the saturated cyclic hydrocarbons containing 3 to 7 carbon atoms, as cyclopropyl, and cyclobutyl, cyclopentyl, cyclohexyl, suberyl, and with short-chain alkyl, hydroxy-lower alkyl, amino-lower alkyl, the substituting group of lower alkoxy-lower alkyl.Word heterocyclic radical refers to saturated or unsaturated (but not being aromatic) four, five, six or seven-membered ring, wherein identical or different nitrogen, oxygen or sulphur atom containing one or two, and this four, five, six or seven-membered ring can suitably with the substituting group of short-chain alkyl, lower alkyloxy, such as piperidyl, morpholinyl, thio-morpholinyl, piperazinyl, THP trtrahydropyranyl, dihydro pyranyl, Isosorbide-5-Nitrae-two alkyl, pyrrolidyl, Pyrrolidine base, pyrrolin base, glyoxalidine base, pyrazolidyl and the substitutive derivative with above-mentioned these heterocycles substituent.Word heteroaryl refers to the six-ring containing one to four nitrogen-atoms, benzo six-ring containing one to three nitrogen-atoms, five yuan of aromatic rings containing a Sauerstoffatom or a nitrogen-atoms or a sulphur atom, five-membered ring fused benzene containing a Sauerstoffatom or a nitrogen-atoms or a sulphur atom, containing a Sauerstoffatom and a nitrogen-atoms five yuan of aromatic rings and benzo derivative thereof, five yuan of aromatic rings containing a sulphur atom and a nitrogen-atoms and benzo derivative thereof, five yuan of aromatic rings containing two nitrogen-atoms and benzo derivative thereof, five yuan of aromatic rings containing three nitrogen-atoms and benzo derivative thereof, or tetrazolium basic ring, such as furyl, thienyl, pyrryl, pyridyl, pyrimidyl, indyl, quinolyl, isoquinolyl, imidazolyl, triazinyl, thiazinyl, thiazolyl, isothiazolyl, pyridazinyl, azoles base, different azoles base, 5-oxygen-1,2,4- di azoly, 5-oxygen-1,2,4-thiadiazolyl group, 5-sulphur-1,2,4- di azoly, 2-oxygen-1,2,3,5- thiadiazolyl group, wherein above-mentioned ring can with substituting group short-chain alkyl, lower alkenyl, amino, amino-lower alkyl, halogen, hydroxyl, lower alkyloxy, trifluoromethoxy, carboxyl, amido, thio acylamino, amidino groups, lower alkyloxy-carbonyl, cyano group, hydroxy-lower alkyl, lower alkoxy-lower alkyl or another aromatic nucleus or heterocyclic ring.Word aryl refer to do not have substituted radical or with the aromatic ring of one, two or three substituent 6 to 10 carbon atom as phenyl ring or naphthalene nucleus, described substituted radical is aryl, halogen, hydroxyl, short-chain alkyl, lower alkenyl, lower alkinyl, lower alkyloxy, lower alkynyl-lower alkoxy, hydroxy-lower alkyl, hydroxy-lower alkenyl, hydroxy-lower alkyl-lower alkinyl, lower alkoxy-lower alkyl, lower alkoxy-lower alkoxyl group, trifluoromethyl, trifluoromethoxy, cycloalkyl, hydroxy-cycloalkyl, heterocyclic radical or heteroaryl.
Second aspect present invention relates to formula (II) peptide derivative or its steric isomer or its pharmaceutical salts,
R-(CH 2) i-CO-(CH 2) j-AA 1(M)-AA 2-AA 3-OH formula (II)
Wherein R, AA 1, AA 2, AA 3, i, j are with the definition in above-mentioned formula (I), and M is as AA 1modification group in structure in atom N, can be short-chain alkyl, lower alkenyl, lower alkinyl, short chain alkyloyl, lower alkyloxy ,-CO 2short-chain alkyl ,-short-chain alkyl-CO 2short-chain alkyl ,-CONH 2,-CONH short-chain alkyl ,-CON [short-chain alkyl] 2.Wherein expression lower alkyloyl refers to ethanoyl, propionyl, butyryl radicals, isobutyryl.
The invention still further relates to formula (I) or formula (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts as the application for the treatment of with the medicine of endothelin effect diseases related.
Term used herein " peptide derivative steric isomer " refers to its corresponding D-or L-steric configuration.
Formula (I) compound can by scheme 1, scheme 2 or the synthesis of scheme 3 method:
i：DMF-DCM，NMM，DCC-HOBt，ii：1M NaOH/MeOH，
Iii:10% citric acid
Scheme 1
In scheme 1, R-CO-AA 1-OH compound (wherein R, AA 1as defined above) and AA 2-OP (wherein AA 2for cis or trans Hyp derivative, P is C 1-4alkyl, it can be selected from methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, preferable methyl and ethyl) react in DMF-DCM, NMM and DCC-HOBt, generate R-CO-AA 1-AA 2-OP compound (wherein R, AA 1, P is as defined above, AA 2for cis or trans Hyp derivative).By R-CO-AA 1-AA 2the saponification in 1M NaOH/MeOH solution of-OP compound, then uses 10% acidified with citric acid, generates R-CO-AA 1-AA 2-OH compound (wherein R, AA 1, AA 2as defined above).By formula R-CO-AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, production R-CO-AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P is as defined above) and compound.Then by gained R-CO-AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, production R-CO-AA 1-AA 2-AA 3-OH.
R-CH 2-CO-AA 1-AA 2-AA 3-OH
I:Et 3n, ii:DCM, iii:1M NaOH/MeOH, iv:10% citric acid,
v：DCM，DMF，DCC/HOBt，NMM
Scheme 2
In scheme 2, AA 1-OP compound (wherein AA 1as defined above, P is C 1-4alkyl, it can be selected from methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, preferable methyl and ethyl) with bromoacetyl bromide at Et 3compd B r-CH is generated in N and DCM 2-CO-AA 1-OP (wherein AA 1, P is as defined above).By compd B r-CH 2-CO-AA 1-OP and R-H (R is as defined above) are at Et 3compound R-CH is generated in N and DCM 2-CO-AA 1-OP (wherein AA 1, P, R are as defined above).By compound R-CH 2-CO-AA 1-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-AA 1-OH (wherein AA 1, R is as defined above).By compound R-CH 2-CO-AA 1-OH and AA 2-OP (wherein AA 2as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-AA 1-AA 2-OP (wherein R, AA 1, AA 2, P is as defined above).By compound R-CH 2-CO-AA 1-AA 2-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-AA 1-AA 2-OH (wherein R, AA 1, AA 2as defined above).By compound R-CH 2-CO-AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P is as defined above).Then by gained R-CH 2-CO-AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates R-CH 2-CO-AA 1-AA 2-AA 3-OH.
I:Et 3n, ii:DCM, iii:NaOH/MeOH, iv:10% citric acid, v: acid anhydrides,
vi：DCM，DMF，DCC-HOBt，NMM
Scheme 3
In scheme 3, compound R-H (wherein R is as defined above) and bromoacetyl bromide are at Et 3compd B r-CH is generated in N and DCM 2-CO-R (wherein AA 1, R is as defined above).By compd B r-CH 2-CO-R and AA 1-OP (wherein AA 1, P is as defined above) and at Et 3compound R-CO-CH is generated in N and DCM 2-AA 1-OP (wherein AA 1, P, R are as defined above).By compound R-CO-CH 2-AA 1-OP saponification in 1M NaOH/MeOH solution, then uses acidified with citric acid, generates compound R-CO-CH 2-AA 1-OH (wherein AA 1, R is as defined above).By compound R-CO-CH 2-AA 1-OH and acid anhydrides (as two-tertiary butyl carbonic anhydride) generate R-CO-CH in the basic conditions 2-(K) AA 1-OH (wherein AA 1, R is as defined above, and K is amino protecting group, as Boc etc.).By R-CO-CH 2-(K) AA 1-OH and AA 2-OP (wherein AA 2, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CO-CH 2-(K) AA 1-AA 2-OP (wherein R, AA 1, AA 2, K, P are as defined above).By compound R-CO-CH 2-(K) AA 1-AA 2-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(K) AA 1-AA 2-OH (wherein R, AA 1, AA 2, K is as defined above).By compound R-CH 2-CO-(K) AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(K) AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P, M are as defined above).By gained R-CH 2-CO-(K) AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(K) AA 1-AA 2-AA 3-OH.Then by compound R-CH 2-CO-(K) AA 1-AA 2-AA 3compound R-CH is obtained after-OH deprotection 2-CO-AA 1-AA 2-AA 3-OH.
And the preparation method of Hyp derivative is as follows:
Wherein R afor amino protecting group, P is carboxyl-protecting group, and itself and formula (6) compound obtain structural formula (5).
G-W-R 2formula (6)
Wherein G is reactive residue, preferred halogen atom; W is-CO-,-CH 2-,-CO-NH-; R 2as structural formula (5) definition.
The preparation method of another kind of Hyp derivative:
formula (7)
Hydroxyl is converted to the method for amino reaction reference literature [1] (Ko, S.Y.J.Org.Chem.2002,67:2689), wherein R afor amino protecting group, P is carboxyl-protecting group, and formula (7) and formula (8) compound obtain structural formula (5).
G-Z-R 2formula (8)
Wherein G is reactive residue, preferred carboxyl, sulfonic group, halogen atom; Z is-O-,-NH-,-CH 2-, a key; R 2as structural formula (5) definition.
Formula (II) compound can by scheme 4, scheme 5 or the synthesis of scheme 6 method:
I:Et 3n, ii:DCM, iii:NaOH/MeOH, iv:10% citric acid,
v：DCM，DMF，DCC-HOBt，NMM
Scheme 4
In scheme 4, (M) AA 1-OP compound (wherein AA 1as defined above, P is C 1-4alkyl, it can be selected from methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, preferable methyl and ethyl, and M is AA 1amido modified group, can be C 1-4alkyl, is selected from methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, preferable methyl, ethyl) with bromoacetyl bromide at Et 3compd B r-CH is generated in N and DCM 2-CO-(M) AA 1-OP (wherein AA 1, P, M are as defined above).By compd B r-CH 2-CO-(M) AA 1-OP and R-H (R is as defined above) are at Et 3compound R-CH is generated in N and DCM 2-CO-(M) AA 1-OP (wherein AA 1, P, R, M are as defined above).By chemical combination R-CH 2-CO-(M) AA 1-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-OH (wherein AA 1, P, R, M are as defined above).By compound R-CH 2-CO-(M) AA 1-OH and AA 2-OP (wherein AA 2as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(M) AA 1-AA 2-OP (wherein AA 1, AA 2, P, R, M are as defined above).By compound R-CH 2-CO-(M) AA 1-AA 2-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-AA 2-OH (wherein R, AA 1, AA 2, M is as defined above).By compound R-CH 2-CO-(M) AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P, M are as defined above).Then by gained R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates R-CH 2-CO-(M) AA 1-AA 2-AA 3-OH.
I:Et 3n, ii:DCM, iii:NaOH/MeOH, iv:10% citric acid,
V: acid anhydrides or carboxylic acid halides, vi:DCM, DMF, DCC-HOBt, NMM
Scheme 5
In scheme 5, compound R-H (wherein R is as defined above) and bromoacetyl bromide are at Et 3compd B r-CH is generated in N and DCM 2-CO-R (wherein AA 1, R is as defined above).By compd B r-CH 2-CO-R and AA 1-OP (wherein AA 1, P is as defined above) and at Et 3compound R-CO-CH is generated in N and DCM 2-AA 1-OP (wherein AA 1, P, R are as defined above).By compound R-CO-CH 2-AA 1-OP saponification in 1M NaOH/MeOH solution, then uses acidified with citric acid, generates compound R-CO-CH 2-AA 1-OH (wherein AA 1, R is as defined above).By compound R-CO-CH 2-AA 1-OH generates R-CO-CH in the basic conditions with acid anhydrides or carboxylic acid halides 2-(M) AA 1-OH (wherein AA 1, R is as defined above, and now M is selected from ethanoyl, propionyl, butyryl radicals, isobutyryl, preferred ethanoyl, propionyl).By R-CO-CH 2-(M) AA 1-OH and AA 2-OP (wherein AA 2, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CO-CH 2-(M) AA 1-AA 2-OP (wherein R, AA 1, AA 2, M, P are as defined above).By compound R-CO-CH 2-(M) AA 1-AA 2-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-AA 2-OH (wherein R, AA 1, AA 2, M is as defined above).By compound R-CH 2-CO-(M) AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P, M are as defined above).By gained R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-AA 2-AA 3-OH.
I:Et 3n, ii:DCM, iii:NaOH/MeOH, iv:10% citric acid,
v：DCM，DMF，DCC-HOBt，NMM
Scheme 6
In scheme 6, compound R-H (wherein R is as defined above) and bromoacetyl bromide are at Et 3compd B r-CH is generated in N and DCM 2-CO-R.By compd B r-CH 2-CO-R and (M) AA 1-OP (wherein AA 1, P is as defined above, and now M is C 1-4alkyl, it can be selected from methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl, preferable methyl, ethyl) at Et 3compound R-CH is generated in N and DCM 2-CO-(M) AA 1-OP.By compound R-CH 2-CO-(M) AA 1-OP saponification in 1M NaOH/MeOH solution, then uses acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-OH.By R-CH 2-CO-(M) AA 1-OH and AA 2-OP (wherein AA 2, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(M) AA 1-AA 2-OP (wherein R, AA 1, AA 2, M, P are as defined above).By compound R-CH 2-CO-(M) AA 1-AA 2-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-AA 2-OH.By compound R-CH 2-CO-(M) AA 1-AA 2-OH and AA 3-OP (wherein AA 3, P is as defined above) react in DMF-DCM, NMM and DCC-HOBt, generate compound R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP (wherein R, AA 1, AA 2, AA 3, P, M are as defined above).By gained R-CH 2-CO-(M) AA 1-AA 2-AA 3-OP saponification in 1M NaOH/MeOH solution, then uses 10% acidified with citric acid, generates compound R-CH 2-CO-(M) AA 1-AA 2-AA 3-OH.
According to the present invention, formula (I) or formula (II) peptide derivative or its steric isomer or its pharmaceutical salts comprise one sequence:
Sequence 1:R-CH 2-CO-AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 2:R-CH 2-CO-AA 1-AA 2-D-Trp-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 3:R-CO-CH 2-AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 4:R-CO-CH 2-AA 1-AA 2-D-Trp-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 5:R-CO-AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
Sequence 6:R-CO-AA 1-AA 2-D-Trp-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
Sequence 7:R-CH 2-CO-(M) AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH 2-；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 8:R-CH 2-CO-(M) AA 1-AA 2-D-Trp-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH 2-；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 9:R-CO-CH 2-(M) AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH-、CH 3CO-，CH 3CH 2CO-；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
Sequence 10:R-CO-CH 2-(M) AA 1-AA 2-D-Trp-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH-、CH 3CO-，CH 3CH 2CO 2-；
AA 1＝Ser、Thr、Phe、Cys、Gln、Asn、Ala、Ile、Leu、Nle、Val；
AA 2＝D-Trp、D-Pro、D-cis-Hyp、D-tran-Hyp、L-cis-Hyp、L-tran-Hyp、
According to the present invention, the steric isomer of formula (I) or formula (II) peptide derivative refers to its corresponding D-or L-steric configuration.
According to the present invention, formula (I) or formula (II) peptide derivative or its steric isomer or its pharmaceutical salts demonstrate excellent results in the anti-ET1 receptor model of animal, therefore can be used as cardiovascular and cerebrovascular medicine for animal, be preferred for Mammals, be more preferably for people.
The present invention also relates on the other hand and containing as at least one formula (I) of the effective dose of activeconstituents or the pharmaceutical composition of (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts and customary pharmaceutical excipients or assistant agent.Pharmaceutical composition can be prepared according to methods known in the art.During for this object, if needed, formula (I) or (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts and one or more solids or liquid pharmaceutical excipients and/or assistant agent can be combined, make the suitable administration form or dosage form that can be used as people.
The present invention relates to prevention on the other hand or treats the method for the disease relevant with endothelin and prostate gland, and the method comprises the formula (I) or formula (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts that give bacterium.
The present invention also relates on the other hand formula (I) or formula (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts purposes in the medicine for the preparation of prevention or the treatment disease relevant with endothelin and prostate gland.
Formula (I) or formula (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts or the pharmaceutical composition containing them can administrations in a unit, route of administration can be enteron aisle or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.Form of administration is tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc. such as.Can be ordinary preparation, sustained release preparation, controlled release preparation and various particulate delivery system.In order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.About the example of carrier, as thinner and absorption agent, as starch, dextrin, lactose, N.F,USP MANNITOL, sucrose, glucose, calcium sulfate, sodium-chlor, urea, calcium carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent, as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, as dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, as talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet can also be made coating tablet further, as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.In order to administration unit is made pill, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, rice paste or batter etc.; Disintegrating agent, as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.In order to administration unit is made suppository, various carrier well known in the art can be widely used.Example about carrier is, the ester, gelatin, semi-synthetic glyceryl ester etc. of such as polyoxyethylene glycol, Yelkin TTS, theobroma oil, higher alcohols, higher alcohols.In order to administration unit is made capsule, effective constituent formula (I) or (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts are mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard capsule or soft capsule.Also effective constituent formula (I) or (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts can be made microcapsule, be suspended in aqueous medium and make suspensoid, also can load in hard capsule or make injection application.In order to administration unit is made injection preparation, as solution, emulsion, lyophilized injectable powder and suspensoid, all thinners that this area is conventional can be used, as the isooctadecanol of water, ethanol, polyoxyethylene glycol, 1,3-PD, ethoxylation, the isooctadecanol, Polyoxyethylene Sorbitol Fatty Acid Esters etc. of polyoxy base.In addition, in order to prepare isotonic injection liquid, appropriate sodium-chlor, glucose or glycerine can be added in injection preparation, in addition, conventional solubility promoter, buffer reagent, pH adjusting agent etc. can also be added.
In addition, as needs, also tinting material, sanitas, spices, correctives, sweeting agent or other material can be added in pharmaceutical preparation.
The dosage of formula (I) or formula (II) peptide derivative and/or its steric isomer and/or its pharmaceutical salts depends on many factors, such as to prevent or the character of disease therapy and severity, the sex of patient or animal, age, body weight and individual reaction, particular compound used, route of administration and administration number of times etc.Above-mentioned dosage can single dose form or be divided into several, such as two, three, four dosage forms for administration.
The abbreviation used in the present invention has implication below:
Ala L-Ala,
Asn l-asparagine,
DAD Isosorbide-5-Nitrae-dioxy-8-azaspiro [4,5] decane,
DBA Di-n-Butyl Amine,
DIC N, N '-diisopropylcarbodiimide
DCM methylene dichloride,
DMF dimethyl formamide,
Diox Isosorbide-5-Nitrae-dioxane
Et 3n triethylamine,
GABA γ-aminobutyric acid,
Gly glycine,
Gln glutamine,
Hyp oxyproline
HIM hexamethylene imine,
HMPA hexylmethylphosphoramide,
HOBt I-hydroxybenzotriazole,
Ile Isoleucine,
Leu leucine,
MeVal methylvaline,
Nle nor-leucine
Mob β-piperonyl L-Ala,
MOP morpholine,
NMM N-methylmorpholine,
Phe phenylalanine,
Phg phenylglycocoll,
Pro proline(Pro),
Pya β-pyriylalanine,
Ser Serine,
TFA trifluoroacetic acid
Thr Threonine,
Trp tryptophane,
Val α-amino-isovaleric acid,
In the present invention, all amino acid configuration, except being labeled as D-type, are L-type.
Embodiment
Embodiment
The following examples and biological activity test are used for further illustrating the present invention, but this and do not mean that any limitation of the invention.
Embodiment melting point compound is measured by RY-1 type melting point apparatus, and temperature is not calibrated; 1h NMR collection of illustrative plates is measured by Bruker ARX 400 type or US Varian Unity Inova 600 type nuclear magnetic resonance spectrometer; FAB mass spectrum is measured by Zabspect high resolution magnetic mass spectrometer; Ultimate analysis is measured by CarloErba 1106 type elemental analyser; UV spectrum is measured by UV-260 ultraviolet-visible pectrophotometer; Infrared spectra is measured by Magna IRTM 550 radar stealthy materials.Embodiment is prepared reaction reagent used and is commercial prod.
Embodiment 1:DBA-CH 2-CO-Leu-D-Trp-D-Phe (4-Cl)-OH
1.1 Br-CH 2-CO-LeuOCH 3(aa 1)
Bromoacetyl bromide 1.00ml (11.56mmol) is dissolved in as in 50ml there-necked flask in 5ml DCM, nitrogen protection.Under-78 DEG C of stirrings, slowly drip DCM (20ml) solution of 2.00g (11.01mmol) leucine methyl ester hydrochloride and 3.13ml (22.57mmol) triethylamine.-78 DEG C of stirring reaction 45min, stirring at room temperature 30min.Be poured into by solution in frozen water, use separating funnel layering, separate the DCM layer of lower floor, then wash 2 times with 1M HCl, saturated NaCl washs 3 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, obtains pale yellow oil 2.93g.
1.2 DBA-CH 2-CO-LeuOH(aa 2)
By 2.93g (11.56mmol) AA 1be dissolved in 50ml acetonitrile, under ice bath, add 1.95ml (11.56mmol) Di-n-Butyl Amine and 1.60ml (11.56mmol) triethylamine, be heated to 40 DEG C of reactions, TLC detects, and after question response is complete, is spin-dried for solvent, add DCM, wash 1 time with water, saturated NaHCO 3wash 3 times, saturated common salt water washing 2 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, and product is separated (DCM: MeOH=100: 1) through silica gel chromatographic column and obtains pale tan oil 2.08g.By this oily matter 25ml dissolve with methanol, 14.55ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 1.89g.
1.3 DBA-CH 2-CO-Leu-D-Trp-D-Phe(4-Cl)-OH
By 1.89g (6.29mmol) AA 2be dissolved in 10ml DCM, add 1.60g (6.29mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.89g HOBt (6.60mmol).N-methylmorpholine 0.69ml (6.28mmol) and 0.83g DIC (6.60mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 2.83g.By this white solid 25ml dissolve with methanol, 11.59ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 2.61g.This solid is dissolved in 10ml DCM, adds 1.34g (5.36mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.76g HOBt (5.63mmol).N-methylmorpholine 0.59ml (5.36mmol) and 0.71g DIC (5.63mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains white solid 3.15g.By this white solid 25ml dissolve with methanol, 9.46ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains white solid 2.72g.
MS：[M+H]＝668.3。
Embodiment 2:DBA-CO-CH 2-Leu-D-Trp-D-Phe (4-Cl)-OH
2.1 DBA-CO-CH 2-Br(bb1)
Bromoacetyl bromide 2.56ml (29.69mmol) is dissolved in as in 100ml there-necked flask in 10ml DCM, nitrogen protection.Under-78 DEG C of stirrings, slowly drip DCM (50ml) solution of 5ml (29.69mmol) Di-n-Butyl Amine and 8.44ml (60.83mmol) triethylamine.-78 DEG C of stirring reaction 45min, stirring at room temperature 30min.Be poured into by solution in frozen water, use separating funnel layering, separate the DCM layer of lower floor, then wash 2 times with 1M HCl, saturated NaCl washs 3 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, obtains pale yellow oil 6.45g.
2.2 DBA-CO-CH 2-(Boc)Leu-OH(bb2)
6.45g (25.78mmol) bb2 is dissolved in 100ml acetonitrile, 4.68g (25.78mmol) leucine methyl ester hydrochloride and 7.34ml (52.85mmol) triethylamine is added under ice bath, be heated to 40 DEG C of reactions, TLC detects, after question response is complete, is spin-dried for solvent, adds DCM, wash 1 time with water, saturated NaHCO 3wash 3 times, saturated common salt water washing 2 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, and product is separated (DCM: MeOH=100: 1) through silica gel chromatographic column and obtains pale tan oil 2.43g.By this oily matter 25ml dissolve with methanol, add 15.84ml 1M sodium hydroxide, stirred at ambient temperature in ice-water bath, TLC detects, and after question response is complete, adds 2.16g (11.59mmol) Boc under ice bath 2o, room temperature maintains pH=9 reaction, and TLC detects, and add ethyl acetate after question response is complete and extract, saturated NaCl washs 3 times, and product is separated (PE: EtOAc=5: 1) through silica gel chromatographic column, obtains white solid 2.47g.
2.3 DBA-CO-CH 2-Leu-D-Trp-D-Phe(4-Cl)-OH
2.47g (6.18mmol) bb2 is dissolved in 10ml DCM, adds 1.57g (6.18mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.88g HOBt (6.49mmol).N-methylmorpholine 0.68ml (6.18mmol) and 0.82g DIC (6.49mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=6: 1) through silica gel chromatographic column, obtains white solid 3.34g.By this white solid 25ml dissolve with methanol, in ice-water bath, add 11.40ml 1M sodium hydroxide, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 7, revolve methyl alcohol, then use 1%-30% acidified with citric acid to pH 2-3, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 3.18g.This solid is dissolved in 20ml DCM, adds 1.36g (5.42mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.77g HOBt (5.69mmol).N-methylmorpholine 0.60ml (5.42mmol) and 0.72g DIC (5.69mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains white solid 3.73g.By this white solid 30ml dissolve with methanol, 10.0ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 7, revolve methyl alcohol, then use 1%-30% acidified with citric acid to extract to pH 2-3, DCM, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 3.29g.To add in 35ml trifluoroacetic acid and 35ml meta-cresol under solid ice bath, stirring at room temperature, TLC detects in batches, and after question response is complete, decompression removing trifluoroacetic acid, adds DCM and water, use Na 2cO 3adjust pH is 7.Separate ester layer drying, filtering siccative, concentrated filtrate, obtain incarnadine oily matter, product is separated (DCM: MeOH=40: 1) through silica gel chromatographic column, obtains white solid 2.23g.
MS：[M+H]＝668.6。
Embodiment 3:DBA-CO-CH 2-Leu (CH 3)-D-Trp-D-Phe (4-Cl)-OH
3.1 (CH 3)LeuOCH 3·HCl(cc1)
5g (0.02mol) tertbutyloxycarbonyl leucine is dissolved in 50ml tetrahydrofuran (THF) and 5ml DMF, under ice bath, adds 2.75g NaH (70%) in batches, after stirring 30min, add 10ml methyl iodide, be heated to 80 DEG C and stir 24h.Decompression removing tetrahydrofuran (THF), adds ethyl acetate, washes 1 time with water, saturated NaHCO 3wash 1 time, saturated NaCl washs 1 time.With anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (PE: EtOAc=25: 1) through silica gel chromatographic column, obtains colorless oil 4.1g.To add 50ml 4M HCl/Diox solution under this oily matter condition of ice bath, stirring at room temperature, TLC detects, and after question response is complete, decompression removing HCl/Diox solution, uses methanol-diethyl ether recrystallization, obtain white crystal 2.93g.
3.2 DBA-CO-CH 2-(CH 3)Leu-OH(cc2)
2.93g (14.97mmol) cc1 and 3.75g (14.97mmol) bb1 is dissolved in 50ml acetonitrile, 4.26ml (30.69mmol) triethylamine is added under ice bath, 40 DEG C of stirrings, TLC detects, after question response is complete, decompression removing acetonitrile, adds DCM, wash 1 time with water, saturated NaHCO 3wash 3 times, saturated common salt water washing 2 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, and product is separated (DCM: MeOH=100: 1) through silica gel chromatographic column and obtains pale tan oil 1.62g.By this oily matter 30ml dissolve with methanol, 10.0ml1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=100: 1) through silica gel chromatographic column, obtains colorless oil 1.38g.
3.3 DBA-CO-CH 2-(CH 3)Leu-D-Trp-D-Phe(4-Cl)-OH
By 1.38g (4.39mmol) cc2 be dissolved in 10ml DCM, add 1.12g (4.39mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.63g HOBt (4.61mmol).N-methylmorpholine 0.48ml (4.39mmol) and 0.58g DIC (4.61mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 1.74g.By this white solid 20ml dissolve with methanol, 6.93ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 1.62g.This solid is dissolved in 10ml DCM, adds 0.81g (3.24mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.46g HOBt (3.40mmol).N-methylmorpholine 0.36ml (3.24mmol) and 0.43g DIC (3.40mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 1.91g.By this white solid 15ml dissolve with methanol, 5.60ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains solid 1.46g.
MS：[M+H]＝682.5。
Embodiment 4:DBA-CO-CH 2-(Ac) Leu-D-Trp-D-Phe (4-Cl)-OH
By 3.1g (10mmol) DBA-CO-CH 2-LeuOCH 3(preparation method is shown in embodiment 2) uses 50ml dissolve with methanol, adds 20.5ml 1M sodium hydroxide, stirred at ambient temperature in ice-water bath, and TLC detects, and after question response is complete, adds 1.40ml (15mmol) Ac under ice bath 2o, room temperature maintains pH=9 reaction, and TLC detects, after question response is complete, be acidified to pH about 1 with concentrated hydrochloric acid, revolve methyl alcohol, add ethyl acetate and extract, saturated NaCl washs 3 times, and product is separated (PE: EtOAc=5: 1) through silica gel chromatographic column, obtains oily matter 2.05g.This oily matter is dissolved in 20mlDCM, adds 0.74g (2.93mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.42g HOBt (3.07mmol).N-methylmorpholine 0.32ml (2.93mmol) and 0.39g DIC (3.07mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=6: 1) through silica gel chromatographic column, obtains white solid 1.56g.By this white solid 15ml dissolve with methanol, 5.74ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 2-3, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 1.49g.This solid is dissolved in 20ml DCM, adds 0.69g (2.75mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.39g HOBt (2.88mmol).N-methylmorpholine 0.30ml (2.75mmol) and 0.36g DIC (2.88mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains white solid 1.79g.By this white solid 15ml dissolve with methanol, 5.0ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 2-3, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=40: 1) through silica gel chromatographic column, obtains white solid 1.40g.
MS：[M+H]＝710.7。
Embodiment 5:DBA-CH 2-CO-(CH 3) Leu-D-Trp-D-Phe (4-Cl)-OH
5.1 Br-CH 2-CO-(CH 3)LeuOCH 3(dd1)
Bromoacetyl bromide 4.3ml (25.6mmol) is dissolved in as in 100ml there-necked flask in 10ml DCM, nitrogen protection.Under-78 DEG C of stirrings, slowly drip DCM (50ml) solution of 5.00g (25.6mmol) cc1 and 7.28ml (52.48mmol) triethylamine.-78 DEG C of stirring reaction 45min, stirring at room temperature 30min.Be poured into by solution in frozen water, use separating funnel layering, separate the DCM layer of lower floor, then wash 2 times with 1M HCl, saturated NaCl washs 3 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, obtains pale yellow oil 8g.
5.2 DBA-CH 2-CO-(CH 3)Leu-OH(dd2)
By 8g (28.6mmol) AA 1be dissolved in 60ml acetonitrile, under ice bath, add 4.8ml (28.6mmol) Di-n-Butyl Amine and 3.97ml (28.6mmol) triethylamine, be heated to 40 DEG C of reactions, TLC detects, and after question response is complete, is spin-dried for solvent, add DCM, wash 1 time with water, saturated NaHCO 3wash 3 times, saturated common salt water washing 2 times, uses anhydrous MgSO 4dry.Filtering siccative, is spin-dried for solvent, and product is separated (DCM: MeOH=100: 1) through silica gel chromatographic column and obtains pale tan oil 7.47g.By this oily matter 120ml dissolve with methanol, 46.61ml1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=20: 1) through silica gel chromatographic column and obtains solid 4.86g.
5.3 DBA-CH 2-CO-(CH 3)Leu-D-Trp-D-Phe(4-Cl)-OH
2g (6.36mmol) dd2 is dissolved in 10ml DCM, adds 1.62g (6.36mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.90g HOBt (6.68mmol).N-methylmorpholine 0.70ml (6.36mmol) and 0.84g DIC (6.68mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, wash with water once, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 2.90g.By this white solid 25ml dissolve with methanol, 11.55ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 2.53g.This solid is dissolved in 10ml DCM, adds 1.67g (5.07mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.72g HOBt (5.32mmol).N-methylmorpholine 0.56ml (5.07mmol) and 0.67g DIC (5.32mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains white solid 3.10g.By this white solid 25ml dissolve with methanol, 9.0ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains white solid 2.10g.
MS：[M+H]＝681.5。
Embodiment 6:
6.1 (ee1) synthesis
Be dissolved in 100ml DCM by trans for 5g (20.39mmol) tertbutyloxycarbonyl hydroxyproline methyl ester, add 1.97ml (24.47mmol) pyridine, ice bath adds 4.66g (24.47mmol) under stirring in batches.Maintenance ice bath reacts, and TLC detects, and after question response is complete, decompression removing DCM, adds ethyl acetate, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=10: 1) through silica gel chromatographic column, obtains white solid 6.83g.This white solid is dissolved in 50ml DMF, add 1.29g (19.82mmol) sodiumazide, be heated to 50 DEG C of stirrings, TLC detects, after question response is complete, and decompression removing DMF, add ethyl acetate, wash 2 times with water, saturated NaCl washs 1 time, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains 4.29g colorless oil.This oily matter is dissolved in 20ml ethyl acetate, add 0.25gPd-C, lead to hydrogen stirring at room temperature to reaction system, stir 10h, filter out Pd-C, wash 1 time with water, adjust pH about 3 with 0.2M HCl, with the raw material that ether extraction unreacted is complete, then adjust pH about 9, extract product with DCM, obtain oily matter 2.56g.This oily matter is dissolved in 10ml DCM, adds 1.84g (10.48mmol) 3-indolyl acetic acid, dissolve with 1ml DMF, then add 1.49g HOBt (11.00mmol).1.39g DIC (11.00mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, wash with water once, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains incarnadine oily matter, adds sherwood oil under ice bath, separates out solid, and filter collection is dry obtains solid 3.49g.To add in 20ml trifluoroacetic acid and 20ml meta-cresol under solid ice bath, stirring at room temperature, TLC detects in batches, and after question response is complete, decompression removing trifluoroacetic acid, adds DCM and water, use Na 2cO 3adjust pH is 9.Separate ester layer drying, filtering siccative, concentrated filtrate, obtain incarnadine oily matter, product is separated (DCM: MeOH=40: 1) through silica gel chromatographic column, obtains white solid 1.31g.
6.2 (ee2) synthesis
1.31g (4.35mmol) ee1 is dissolved in 10ml DCM, adds 1.25g (4.35mmol) DBA-CO-Leu-OH, dissolve with 1ml DMF, then add 0.62g HOBt (4.57mmol).0.58g DIC (4.57mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 1.73g.By this white solid 15ml dissolve with methanol, 6.5ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 2-3, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains white solid 1.01g.
MS：[M+H]＝556.7。
Embodiment 7:
0.50g (0.90mmol) ee2 is dissolved in 5ml DCM, adds 0.20g (0.90mmol) D-4-chlorophenylalanine methyl ester hydrochloride, dissolve with 0.5ml DMF, then add 0.13g HOBt (0.95mmol).N-methylmorpholine 0.10ml (0.90mmol) and 0.12g DIC (0.95mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 0.60g.By this white solid 5ml dissolve with methanol, 1.5ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 2-3, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains white solid 0.45g.
MS：[M+H]＝737.7。
Embodiment 8:
8.1 (ff1) synthesis
With D-cis tertbutyloxycarbonyl hydroxyproline methyl ester for raw material, corresponding operating is with embodiment 6.
8.2 (ff2) synthesis
0.87g (2.90mmol) ff1 is dissolved in 10ml DCM, adds 0.90g (2.90mmol) AA 2, dissolve with 1ml DMF, then add 0.50g HOBt (3.50mmol).0.40g DIC (3.50mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 1.43g.By this white solid 15ml dissolve with methanol, 5ml1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, product is separated (DCM: MeOH=20: 1) through silica gel chromatographic column, obtains white solid 1.11g.
MS：[M+H]＝570.6。
Embodiment 9:
Take ff2 as raw material (preparation method is shown in embodiment 8), corresponding operating is with embodiment 1.
MS：[M+H]＝751.8。
Embodiment 10:
1.60g (5.30mmol) ff1 is dissolved in 50ml DCM, adds 2.10g (5.30mmol) bb2, dissolve with 1ml DMF, then add 0.90g HOBt (6.40mmol).0.80g DIC (6.40mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use 1%-30% citric acid respectively, saturated NaHCO 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains light red solid 3.25g.By this white solid 25ml dissolve with methanol, in ice-water bath, add 9.7ml 1M sodium hydroxide, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 7, revolve methyl alcohol, then use 1%-30% acidified with citric acid to pH 2-3, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, adds sherwood oil under ice bath, separates out solid, and filter collection is dry obtains solid 3.06g.To add in 60ml trifluoroacetic acid and 60ml meta-cresol under solid ice bath, stirring at room temperature, TLC detects in batches, and after question response is complete, decompression removing trifluoroacetic acid, adds DCM and water, use Na 2cO 3adjust pH is 7.Separate ester layer drying, filtering siccative, concentrated filtrate, obtain incarnadine oily matter, product is separated (DCM: MeOH=20: 1) through silica gel chromatographic column, obtains white solid 1.56g.
MS：[M+H]＝570.7。
Embodiment 11:
With shown compound is raw material (preparation method is shown in embodiment 10), and corresponding operating is with embodiment 2.
MS：[M+H]＝751.8。
Embodiment 12:
12.1 (gg1) synthesis
Trans for 5g (20.39mmol) tertbutyloxycarbonyl hydroxyproline methyl ester is dissolved in 100ml tetrahydrofuran (THF), add 1.40g (40.78mmol) NaH (70%) under ice bath in batches, after stirring 1h, add 3.48g (20.39mmol) pepper methyl chloride, room temperature reaction 24h.Decompression removing tetrahydrofuran (THF), add ethyl acetate, wash 1 time with water, saturated NaCl washs 1 time.With anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (PE: EtOAc=25: 1) through silica gel chromatographic column, obtains colorless oil 3.78g.To add 25ml 4M HCl/Diox solution under this oily matter condition of ice bath, stirring at room temperature, TLC detects, after question response is complete, decompression removing HCl/Diox solution, adds water, with the impurity that ether extraction polarity is little, adjust pH=9, extract with DCM, separate ester layer drying, filtering siccative, concentrated filtrate, product is separated (DCM: MeOH=40: 1) through silica gel chromatographic column, obtains white solid 1.67g.
12.2 (gg2) synthesis
Take gg1 as raw material, corresponding operating is with embodiment 6.
MS：[M+H]＝532.4。
Embodiment 13:
Take gg2 as raw material, corresponding operating is with embodiment 7.
MS：[M+H]＝715.8。
Embodiment 14:
Take gg1 as raw material, corresponding operating is with embodiment 8.
MS：[M+H]＝548.4。
Embodiment 15:
With embodiment 14 product for raw material, corresponding operating is with embodiment 1.
MS：[M+H]＝729.3。
Embodiment 16:
Take gg1 as raw material, corresponding operating is with embodiment 10.
MS：[M+H]＝548.4。
Embodiment 17:
With shown structure is raw material (preparation method is shown in embodiment 16), and corresponding operating is with embodiment 2.
MS：[M+H]＝729.2。
Embodiment 18:DBA-CH 2-CO-Leu-D-Trp-D-Trp-OH
Be that raw material is dissolved in 5ml DCM by 1.00g (3.33mmol) aa2, add 0.85g (3.33mmol) D-trp methyl ester hydrochloride, dissolve with 0.5ml DMF, then add 0.47gHOBt (3.49mmol).N-methylmorpholine 0.37ml (3.33mmol) and 0.44g DIC (3.49mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=3: 1) through silica gel chromatographic column, obtains white solid 1.51g.TLC detects: (methylene dichloride: methyl alcohol 20: 1) Rf=0.3.By this white solid 10ml dissolve with methanol, 6ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Add sherwood oil under ice bath, separate out solid, filter collection is dry obtains solid 1.43g.This solid is dissolved in 10ml DCM, adds 0.75g (2.94mmol) D-trp methyl ester hydrochloride, dissolve with 1ml DMF, then add 0.35g HOBt (2.61mmol).N-methylmorpholine 0.27ml (2.49mmol) and 0.33g DIC (2.61mmol) is added under condition of ice bath.Continue to keep ice bath after 0.1-5 hour, room temperature reaction 1-10 hour.Concentrated, add ethyl acetate, after washing 1 time with water, use saturated NaHCO respectively 33 times are respectively washed, ethyl acetate layer anhydrous sodium sulfate drying 3-6 hour with saturated NaCl.Filtering siccative, concentrated filtrate, obtains colorless oil, and product is separated (PE: EtOAc=4: 1) through silica gel chromatographic column, obtains white solid 1.80g.TLC detects: (methylene dichloride: methyl alcohol 20: 1) Rf=0.5.By this white solid 25ml dissolve with methanol, 5.50ml 1M sodium hydroxide is added in ice-water bath, stirred at ambient temperature, TLC detects, after question response is complete, with 1%-30% acidified with citric acid to pH about 6, revolve methyl alcohol, separate out solid, DCM extracts, after saturated NaCl solution washing, with anhydrous sodium sulfate drying 3-6 hour.Filtering siccative, concentrated filtrate, obtains colorless oil.Product is separated (DCM: MeOH=30: 1) through silica gel chromatographic column, obtains white solid 1.60g.
MS：[M+H]＝673.3。
Embodiment 19:
With gg2 and tryptophan methyl ester hydrochloride for raw material, preparation method is by embodiment 13.
MS：[M+H]＝720.6。
Embodiment 20:
Evaluated biological activity: adopt anti-ET-1 to cause isolated rat aortic rings and shrink experiment
(1) experiment material
Wistar rat: Military Medical Science Institute's Experimental Animal Center
Desk-top self-balancing recorder: Shanghai great Hua instrument plant
ET-1:sigma company
10% salt of wormwood autogamy
Improvement Kreb ' s-Ringer damping fluid: autogamy
(2) experimental technique
The preparation of sample solution:
Quantitatively take sample 2 × 10 -6mol, adds 120 μ l 10% alkaline carbonates and 1-80 μ l DMSO dissolves, and quantitatively dilutes for 5.0ml, preserve as in stock solution refrigerator with improvement Kreb ' s-Ringer damping fluid.Diluting with damping fluid is 10 -6, 10 -7, 10 -8with 10 -9m tetra-concentration.
Prepared by extracorporeal blood vessel:
By rat sacrificed by decapitation, open chest rapidly, win aorta, be placed in the culture dish filling vascular nutrition liquid, remove blood stains, careful separating blood vessel surrounding tissue, be cut into the arterial ring being about 3mm, the stainless steel steel wire being 0.1mm by two diameters respectively carefully penetrates, and makes triangle ring-type.Fill in 37 DEG C of thermostatic baths of 10ml vascular nutrition liquid with being placed on, lower end is fixed, and upper end is logical moves tonotransducer and be connected in desk-top self-balancing recorder, continues to pass into 95% oxygen and 5% carbon dioxide mix gas.Arterial ring load 0.5g, starts administration after stable.
Antagonistic experiment:
First bring out vascular circle with the ET-1 of 10nM amount to shrink, about about 10 minutes, after reaching platform, give 10 -9m sample, observes antagonism shrinking effect.When lower concentration is not imitated, elevated concentrations to 10 gradually -6m.
Result:
NO IC 50(M) ET-1 Ki
Positive control BQ-485 1.23E-07 1.00E-08 2.37E-08
1 7.35E-08 1.00E-08 1.41E-08
2 6.74E-08 1.00E-08 1.29E-08
3 6.58E-08 1.00E-08 1.26E-08
4 3.74E-08 1.00E-08 7.18E-09
5 6.30E-08 1.00E-08 1.21E-08
6 5.11E-08 1.00E-08 9.82E-09

Claims (7)

1. formula (I) peptide derivative or its pharmaceutical salts
R-(CH 2) i-CO-(CH 2) j-AA 1-AA 2-AA 3-OH formula (I)
Wherein, R is hexamethylene imine base or following group:
In structural formula 1, n 1, n 2and n 3be integer 0,1,2 or 3 independently, and wherein n 3≠ 0,0≤n 1+ n 2≤ 4; X 1for NH, O, S or CH 2; X 2for N; Work as n 1=n 2=1, n 3=2, X 1for CH 2, X 2during for atom N, R is abbreviated as ABO;
In structural formula 2, m 1, m 2, m 3and m 4be integer 0,1 or 2 independently, and wherein 0≤m 1+ m 2≤ 4,0≤m 3+ m 4≤ 4; y 1and y 3be CH independently 2, O, S or NH, y 2for N; Work as m 1=m 2=1, m 3=m 4=2, y 1, y 3for O atom, y 2during for atom N, R is abbreviated as DAD; Work as m 1=m 2=1, m 3=m 4=2, y 3for CH 2, y 1for O atom, y 2during for atom N, R is abbreviated as CSO;
In structural formula 3, z is N or CH, k be N, t is H, CH 3, CH 2cH 3or CH 1(CH 3) 2; Or z is O or S, k is that N, t and connecting key thereof do not exist; Work as z=N, when k=N, t=H, R is piperazinyl; Work as z=O, when k=N, t and connecting key thereof do not exist, R is morpholinyl;
In structural formula 4, a and b is integer 0,1,2 or 3 independently; P is atom N; Work as a=3, during b=3, R is abbreviated as DBA;
AA 1for L or D type Leu;
AA 2for L or D type Trp;
AA 3for D type phenylalanine, the phenyl wherein in aromatic nucleus can be selected from halogen, nitro, urea groups, methoxyl group on 2,3,4 or 5, the substituting group of carboxyl and short-chain alkyl monosubstituted or two replace;
0≤i≤4；0≤j≤4；
Condition be i with j different time be 0.
2. formula (II) peptide derivative or its pharmaceutical salts
R-(CH 2) i-CO-(CH 2) j-(M) AA 1-AA 2-AA 3-OH formula (II)
Wherein R, AA 1, AA 2, AA 3, i, j are with the definition in claim 1 Chinese style (I), and M is as AA 1modification group in structure in atom N, can be short-chain alkyl, lower alkenyl, lower alkinyl, short chain alkyloyl, lower alkyloxy ,-CO 2short-chain alkyl ,-short-chain alkyl-CO 2short-chain alkyl ,-CONH 2,-CONH short-chain alkyl ,-CON [short-chain alkyl] 2.
3. the formula (I) described in claim 1 or 2 or formula (II) peptide derivative or its pharmaceutical salts, wherein said formula (I) or formula (II) peptide derivative or its pharmaceutical salts are selected from:
Sequence 1:R-CH 2-CO-AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Leu；
AA 2＝D-Trp；
Sequence 3:R-CO-CH 2-AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
AA 1＝Leu；
AA 2＝D-Trp；
Sequence 7:R-CH 2-CO-(M) AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH 2-；
AA 1＝Leu；
AA 2＝D-Trp；
Sequence 9:R-CO-CH 2-(M) AA 1-AA 2-D-Phe (4-Cl)-OH
R＝DBA，ABO，DAD，HIM，MOP；
M＝CH 3-、CH 3CH-、CH 3CO-，CH 3CH 2CO-；
AA 1＝Leu；
AA 2＝D-Trp。
4. the formula (I) described in claim 1 or 2 or formula (II) peptide derivative or its pharmaceutical salts, wherein said formula (I) or formula (II) peptide derivative or its pharmaceutical salts are selected from:
Compound 1:DBA-CH 2-CO-Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 2:DBA-CO-CH 2-Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 3:DBA-CO-CH 2-Leu (CH 3)-D-Trp-D-Phe (4-Cl)-OH;
Compound 4:DBA-CO-CH 2-(Ac) Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 5:DBA-CH 2-CO-(CH 3) Leu-D-Trp-D-Phe (4-Cl)-OH.
5. pharmaceutical composition, it comprises formula (I) described at least one claim 1 or 2 or formula (II) peptide derivative or its pharmaceutical salts and pharmaceutical carrier or vehicle.
6. the pharmaceutical composition of claim 5, wherein said formula (I) or formula (II) peptide derivative or its pharmaceutical salts are selected from:
Compound 1:DBA-CH 2-CO-Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 2:DBA-CO-CH 2-Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 3:DBA-CO-CH 2-Leu (CH 3)-D-Trp-D-Phe (4-Cl)-OH;
Compound 4:DBA-CO-CH 2-(Ac) Leu-D-Trp-D-Phe (4-Cl)-OH;
Compound 5:DBA-CH 2-CO-(CH 3) Leu-D-Trp-D-Phe (4-Cl)-OH.
7. the formula (I) described in claim 1 or 2 or formula (II) peptide derivative or its pharmaceutical salts purposes in the medicine for the preparation of prevention or the treatment disease relevant with endothelin.
CN200810007611.3A 2008-02-29 2008-02-29 Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof Expired - Fee Related CN101519427B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN200810007611.3A CN101519427B (en) 2008-02-29 2008-02-29 Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN200810007611.3A CN101519427B (en) 2008-02-29 2008-02-29 Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof

Publications (2)

Publication Number Publication Date
CN101519427A CN101519427A (en) 2009-09-02
CN101519427B true CN101519427B (en) 2015-02-25

Family

ID=41080249

Family Applications (1)

Application Number Title Priority Date Filing Date
CN200810007611.3A Expired - Fee Related CN101519427B (en) 2008-02-29 2008-02-29 Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof

Country Status (1)

Country Link
CN (1) CN101519427B (en)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010142058A1 (en) * 2009-06-09 2010-12-16 中国人民解放军军事医学科学院毒物药物研究所 Peptide derivatives with endothelin receptor antagonizing activity, pharmaceutical compositions and uses thereof
JP5279938B1 (en) 2011-11-09 2013-09-04 国立大学法人九州大学 Valuable metal extractant and method for extracting valuable metal using the extractant
JP5279942B1 (en) * 2011-11-09 2013-09-04 国立大学法人九州大学 Cobalt extraction method
JP5367862B2 (en) 2012-03-13 2013-12-11 国立大学法人九州大学 Scandium extractant and method for extracting scandium using the extractant
JP5398885B1 (en) 2012-08-20 2014-01-29 国立大学法人九州大学 Gallium extraction method
JP5734268B2 (en) 2012-12-12 2015-06-17 国立大学法人九州大学 Nickel extraction method
JP5595554B1 (en) 2013-03-18 2014-09-24 国立大学法人九州大学 Method for separating impurities from an acidic solution containing nickel, cobalt and / or scandium
JP5940688B1 (en) 2015-01-20 2016-06-29 国立大学法人九州大学 Zirconium extractant and zirconium extraction method

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1330075A (en) * 2000-06-21 2002-01-09 中国人民解放军军事医学科学院毒物药物研究所 Endothelin antagon
CN1504480A (en) * 2002-11-28 2004-06-16 中国人民解放军军事医学科学院毒物药 Endothelin receptor antagonist of tripeptide

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1330075A (en) * 2000-06-21 2002-01-09 中国人民解放军军事医学科学院毒物药物研究所 Endothelin antagon
CN1504480A (en) * 2002-11-28 2004-06-16 中国人民解放军军事医学科学院毒物药 Endothelin receptor antagonist of tripeptide

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
J.Dong et al.Synthesis and evaluation of new series of peptide-based endothelin receptor antagonists.《The Journal of Peptide Research》.2005,第65卷(第4期),第440-444页. *
梁远军等.内皮素受体肽类拮抗剂的研究进展.《国外医学药学分册》.2000,第27卷(第4期),第217-221页. *
费改顺等.新型肽类化合物对内皮素受体的拮抗作用及其心血管药理活性的评价.《药学学报》.2002,第37卷(第8期),第593-597页. *

Also Published As

Publication number Publication date
CN101519427A (en) 2009-09-02

Similar Documents

Publication Publication Date Title
CN101519427B (en) Peptide derivative with endothelin receptor antagonizing activity, drug composition and application thereof
JP3938690B2 (en) Selective linear peptide having melanocortin-4 receptor (MC4-R) agonist activity
JP4653840B2 (en) Metastin derivatives and uses thereof
KR101424306B1 (en) Metastin derivatives and use thereof
US5502032A (en) Peptides, the preparation and use thereof
EP0832104B1 (en) Dolastatin derivatives, their preparation and use
US20060135436A1 (en) Peptides and methods for the control of obesity
US20030092609A1 (en) Novel antiarrythmic peptides
BG65065B1 (en) Peptide antiangiogenic drugs
US20110118172A1 (en) Metastin derivative and use thereof
KR100286242B1 (en) Dolastatin derivatives
JP2001089499A (en) Peptide-like compound containing rgd sequence and useful as integrin inhibitor
CN111533800B (en) Novel somatotropin-releasing hormone analogous peptide modification and dimerization preparation and application thereof
AU641366B2 (en) Peptide compounds
CA2385659C (en) Novel antiarrhythmic peptides
NO177713B (en) Analogous procedure for the preparation of hemoregulation peptides
US5786335A (en) Sulfhydryl containing peptides for treating vascular disease
US4203975A (en) Psychopharmacological peptides suitable for therapeutic administration
CN100500691C (en) Peptide-like endothelin receptor antagonist with proline or cyclic amino acid on 2-postion
CN113150068B (en) Peptide compound, application thereof and composition containing peptide compound
HU228993B1 (en) Low molecular weight peptide derivatives as inhibitors of the laminin/nidogen interaction, their use in preparation of medicaments, medicaments and process for their preparation
WO2002068457A2 (en) Antiangiogenic peptides derived from endostatin
JP2003089700A (en) Peptide
US20120129774A1 (en) Kinin b1 receptor peptide agonists and uses thereof
EP1152011A1 (en) Peptides inhibiting vascular endothelial cell migration

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20150225

Termination date: 20160229

CF01 Termination of patent right due to non-payment of annual fee