CN1094058A - Brad ykinin antagonists - Google Patents

Brad ykinin antagonists Download PDF

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CN1094058A
CN1094058A CN93114484A CN93114484A CN1094058A CN 1094058 A CN1094058 A CN 1094058A CN 93114484 A CN93114484 A CN 93114484A CN 93114484 A CN93114484 A CN 93114484A CN 1094058 A CN1094058 A CN 1094058A
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dipolymer
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group
bka
peptide
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J·C·彻奥尼斯
J·K·博劳德格特
V·S·古德佛楼
M·V·马拉瑟
L·W·思普鲁斯
E·T·瓦雷
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Cortech Inc
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/18—Kallidins; Bradykinins; Related peptides
    • A—HUMAN NECESSITIES
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    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract

The brad ykinin antagonists of following formula:
(BKA n)(X)(Y)
BKA wherein nBe the brad ykinin antagonists peptide, Y is a pharmacophoric group; And X is that chemistry links BKA nBridged group with the Y component.

Description

Brad ykinin antagonists
The invention relates to have drug action by brad ykinin antagonists (BKA n) component links the heterogeneous dipolymer of forming (het-erodimers) with another kind of different pharmacophoric group component covalency.
In the disclosed brad ykinin antagonists dipolymer of in first to file WO92/17201, having described following type on October 15th, 1992:
BKA wherein nRepresent a kind of brad ykinin antagonists peptide, X is for linking two kinds of BKA nIntermediary's group of component end.BKA nComponent can be identical or different.But also described some and linked BKA by linking radicals X nThe heterogeneous dipolymer of peptide and another kind of different receptor active peptides, for example NK 1Or NK 2Antagonist peptide or μ-opioid receptor agonist peptide.When in close relations between the related activity, these heterogeneous dipolymers are particularly useful.Known in multiple important pathological physiological process, there is the interaction between inflammatory and the neurohumor.For example in tissue injury (accidental and operation back) Secondary cases pain and asthma, all there is this situation.The medium that all exists tissue and blood plasma to produce in both cases (as acts on BK 2The kassinin kinin of acceptor) and the neurone factor such as the P material (NK that produce 1Acceptor) and neurokinin (Neurokinin) A(NK 2Acceptor) complex interactions between.μ-opiates the neuronal acceptor that also has local action can suppress the no matter release of the neuropeptide (P material, neurokinin A, neurokinin B, cholecystokinin, CGRP etc.) of which kind of type behind irriate.
There is not a kind of medicine generally to improve the physiopathology symptom that the interaction between above-mentioned medium or other inflammatory and neurohumor is brought out effectively.The heterogeneous dipolymer that above-mentioned patent application is described is to attempt to solve these problems with the single medicine with two-way choice.Other advantage of heterogeneous dipolymer like this is familiar with by those skilled in the art equally.
Say on the broad sense, the invention relates to a kind of BKA of binding nThe pharmacophoric group of peptide and another kind of non-brad ykinin antagonists and the heterogeneous dipolymer that forms, for example this pharmacophoric group can be the peptide as describing among the WO92/17201, or non-peptide material, and the non-bradykinin action component as the active relevant problem of pain and/or inflammation or other and kassinin kinin there is different effect.The compound of the compound that forms for having " dual function ", it can act on two receptoroids or act on a kind of being subjected to originally and a kind of enzyme.This is not that individual molecule can be simultaneously and two kinds of acceptors or a kind of acceptor and a kind of enzyme effect; Just say this molecule can with a kind of acceptor in two kinds of acceptors or with a receptoroid and/or a kind of enzyme in any acts on mutually.Use this compound of suitable dosage, its overall pharmacological action be equivalent at least two kinds active add and.This compound can be designed to the compound that is kept perfectly, or after being designed to resolve into two isolating molecules, each self-sustaining oneself is independently active.
Heterogeneous dipolymer of the present invention can be represented with the following formula structure.
BKA wherein nBe the brad ykinin antagonists peptide; X is peptide or non-peptide pharmacophoric group for linking group and Y, and it is for non-brad ykinin antagonists and have and act on and BKA nThe acceptor that component is different or the activity of enzyme preferably act on the process with pain or inflammation-related.
Heterogeneous dipolymer of the present invention provides the possibility of wide spectrum ground treatment pain and inflammation.It is generally acknowledged at inflammatory conditions, regardless of its severity, with the treatment of single medicine or medium or explain that the possibility of all clinical symptom of this syndromes is atomic little.Inference according to this, because the effect of above-mentioned bradykinin on the inflammation physiopathology, any complex therapy of treatment inflammatory disease all should comprise the brad ykinin antagonists part of therapeutic action as a whole.Wide spectrum and effective non-specific methods of treatment (for example using steroid in asthma) are though may be that effectively they can cause more serious toxic side effect.
In many cases, known two kinds of important clinical symptoms of medium synergy and the soluble overwhelming majority diseases for the treatment of independently.For example say after asthma and the wound or postoperative pain and P material act on NK 1Acceptor and bradykinin act on BK 2The relation of acceptor.Equally, the neutrophilia elastoser is that a very important inflammation downstream effects factor and bradykinin are an important initiation and lasting inflammatory mediator, and both effects can be counted as collaborative.
Pharmacological active substance is made homogeneity dipolymer (homodimers) to be described in other system of front with the notion of improving characteristics such as metabolic stability, selectivity and receptors bind.The prior art of this respect comprises in order to increase effect effectiveness and/or action time peptide agonists and antagonist dimerization.See Proc.10th American Pep-tide Symp. such as Caporale, Pierce chemical company, Rockford, IL 449-451(1988) and the european patent application No.EP 293130A2 of Rosenblatt etc.Such peptide agonists dimerization thing discloses enkephalin/endorphin (Shimohigashi, Y. etc., BBRC, 146,1109-1115,1987); P material (Higuchi, E.J.P. such as Y., 160,413-416,1989); Bradykinin (Vavrek, R. and Stewart., J.J.Proc.8th Amer.Pept.Symp., 381-384,1983); Neurokinin A and B(Ko-dama, E.J.P. such as H., 151,317-320,1988); Regular Insulin (Roth, FEBS such as R.A., 170,360-364,1984) and anterior chamber's natriuretic peptide (Chino, BBRC such as N., 141,665-672,1986).The antagonist dimerization is disclosed as Rat parathyroid hormone 1-34 (Caproale, Proc.10th Amer.Pept.Symp. such as L.H., 449-451,1987).But document does not disclose the heterogeneous dipolymer that this paper indication is made up of brad ykinin antagonists and different pharmacophoric groups.
Many known brad ykinin antagonists are arranged in the art, wherein anyly all can be used as BKA in the dipolymer of the present invention nComponent.A kind of peptide that is following formula at external effectively brad ykinin antagonists:
D-ARG 0-Arg 1-Pro 2-Hyp 3-Gly 4-Phe 5-Ser 6-D-Phe 7-Leu 8-Arg 9
See Regoli etc., Trends in Pharmacological Science, 11:156-161(1990).This peptide is referred to herein as CP-0088 for simplicity.
CP-0088 is a kind of BKA easy to use n, those skilled in the art can find out that other has or known brad ykinin antagonists peptide also can be used for purpose of the present invention certainly.Various such buffering kinin antagonists peptides are disclosed by nearest patent documentation, wherein anyly all can be used for purpose of the present invention.See as EP-A-0334244(Procter and Gamble) nonapeptide or bigger brad ykinin antagonists peptide are disclosed, wherein some amino-acid residue is modified.EP-A-0370453(Hoechst) and WO.89/01780 and WO 89/01781(Stewart etc.) the brad ykinin antagonists peptide also described.This paper indication dipolymer does not appear in these patent publications.But the peptide of noticing these publications can be used for implementing the present invention.
Any binding radicals X all can be used for the present invention so that BKA nLink together with chemistry or covalent manner with the Y component, as long as this does not influence BKA nGet final product with the activity of Y component.Link group and can be inorganic (as-S-) or organically, and can select concatenating group group so that hydrolysis or otherwise decompose and discharge two kinds of active ingredient BKA in vivo nAnd Y.Perhaps, link group and can make this heterogeneous dipolymer compound that is kept perfectly in use.
Link radicals X easily and can comprise-the S-atom that it is by BKA nSulfydryl on the peptide chain and other pharmacophoric group component reaction generation.This can finish by relating in the peptide chain as the reaction of the sulfydryl of half fat propylhomoserin (Cys) of peptide P end.This may modify initial BKA in the initial stage nPeptide makes it contain the Cys group in the appropriate location of peptide chain.For example CP-0088 can replace Ser with Cys and modifies (CP-0088 of modified is called CP-0126 hereinafter like this) and link easily with sulfydryl and other pharmacophoric group by Cys in the 6-position.
CP-0126 can be represented by the following formula structure:
Figure 931144841_IMG1
Following formula can be expressed as follows with abbreviation:
With Cys as linking the site, link so radicals X comprise half deamination acid sulfydryl-S-.It can be that whole binding group X(is as in the disulfide group dipolymer) or only be the part of X.Like this, for example link group and can comprise one two succinimido alkane as two succinimido hexanes, it is attached at BKA with end nOn the Y component.Applicant's related application also discloses other and has linked group, and they all can be used for this purpose.Other links group X(, and wherein some does not require or does not contain-the S-atom) can derive from following six compounds of group, they are classified as generally that amino acid analogue links agent or based on the binding agent of maleimide.These link agent only is example, does not think that here exhaustive all potential link group:
The first kind (amino acid analogue)
Figure 931144841_IMG2
Amino acid analogue links agent (first kind) can directly be incorporated into BKA nIn the peptide chain, be used for then forming stable or unsettled heterogeneous dipolymer to esterase with paired pharmacophoric group (Y component).In addition, can react with desirable pharmacophoric group, link with the peptide that contains sulfydryl then based on the binding agent of maleimide.At last, any R that contains in these several compounds 3Group is-CO 2The binding agent of H can link the agent reaction with the another kind in these several compounds and to form the esterase instability (be contained R 3The binding agent of=-OH) or to esterase is stable (contain R 3=-NH 2The binding agent) the binding agent, be used to form the peptide/heterogeneous dipolymer of non-peptide of hope then.R 1And R 2Can change, so that the approaching of ester carbonyl group carbon that links group do not hindered fully or obviously hinder, thereby wish the said ester of control hydrolysis rate in vivo.
Some be used for preparing dipolymer of the present invention for linking the BKA that agent is modified nOr pharmacophoric group itself carries newly, and further constitutes one aspect of the present invention.
The Y component of this heterogeneous dipolymer can be the peptide of any non-brad ykinin antagonists or the pharmacophoric group of non-peptide, thereby it has the activity (non-bradykinin) of the different aspect that resists relevant pain and/or inflammatory process forms dual action compounds, and it can act on two classes not isoacceptor or a kind of acceptor and a kind of enzyme independently.For example the Y component is non-peptide μ-opioid receptor agonist, as morphine or its a kind of derivative such as 14-hydroxyl dihydrocodeinone or 14-hydroxyl Hydromorphone.
When wishing inhibitors of cyclooxygenases (COI), INDOMETHACIN can be used as a useful selection of Y component.But other any traditional non-sterols anti-inflammatory drug such as Asprin, Ibuprofen BP/EP, naprosine (naproxyn) etc. all can be used.In this case, heterogeneous dipolymer BKA n/ COI may need to be hydrolyzed so that obtain the COI activity in vivo, because epoxidase is generally considered to be a kind of intracellular enzyme.The neutrophilia elastase inhibitor may be a kind of active ester, as 2-octadecyloxy phenyl acid esters.It also can be used as heteroaryl alkanoic acid ester esterase inhibitor.A kind of elastase inhibitor of neutrophilia preferably that can be used as the Y component is CE-1218 as described below.It is believed that this is a kind of new compound, and further form a feature of the present invention.
The elastase inhibitor of other type also can be used as the Y component, comprises methyl fluoride ketone, phosphoric acid salt, benzoxazoles, beta-lactam etc.
As above-mentioned, the Y component can have the suitable active inhibitor of brad ykinin antagonists that is different from by peptide or non-peptide class and be formed.But the Y component is preferably to having generally or the acceptor or the activated material of enzyme of substantial connection with the bradykinin activity.As about treatment pain or inflammation.Below saying something, BKA is discussed nWith μ-opioid receptor agonist, neutrophilia elastase inhibitor, inhibitors of cyclooxygenases or NK 1Or NK 2Receptor antagonist is under various conditions in conjunction with the theoretical basis of using.
BKA n/ μ-opioid receptor agonist
The nervosa part of the known mediation pain sensation of C-fiber esodic nerve and inflammation.These afferent neurons discharge various neuropeptides be subjected to specificity and nonspecific stimulation in the tissue of central nervous system (CNS) and peripheral nerve domination after.Some neuropeptides comprise: P-material, neurokinin A, neurokinin B, thyrocalcitonin group related peptides (CGRP), cholecystokinin (CCK), vasoactive intestinal polypeptide (VIP), neuropeptide tyrosine and other neurotransmitter.For these complex materials, according to nerve innervate tissue, different C-fibers contains these neuropeptides of different amounts and/or ratio.Shown that all these peptides work in the various nervosa processes that influence numerous disease and clinical syndrome.In fact many pharmaceutical companies and independent studies laboratory are developed as possible medicine with the specific antagonists of these peptides.
These different types of neuronic tangible common traits are that they all have μ-opiate receptor of regulating above-mentioned neuropeptide release.No matter the endogenous brain deltorphin delta still is that the micromolecular compound that injects of other external source such as morphine, 14-hydroxyl Hydromorphone, fentanyl and their derivative can be as μ-opioid receptor agonists of partial (peripheral nerve endings μ-opiate receptor) and CNS, thereby inhibition discharges neuropeptide from periphery C-fiber.Those peptides that this restraining effect and C-fiber are contained and the stimulation that causes them to discharge all have nothing to do.
As a result, BKA nThe heterogeneous dipolymer of/μ-opioid receptor agonist is considered to an important compounds, and they have good overall activity to the integrated status that produces in treatment hormone and the nervosa process especially.Wish that these compounds not only lower or block with the kassinin kinin hormone part of the inflammatory process that is representative, and the block nerves peptide discharges the nervosa part of the inflammation that causes.In addition, the aspect of using a limitation of existing μ-opioid receptor agonist is that they tend to produce calmness, psychiatric disorder and suppress respiratory movement, says nothing of the danger that develops into habituation and/or tolerance the patient who treats with these medicines on one's body in addition.The bad aspect of these of μ-opioid receptor agonist is because they are easy to see through the ability of CNS.But because BKA nThe high-cation characteristic, BKA nThe heterogeneous dipolymer of/μ-opioid receptor agonist would not penetrate CNS.As a result, the activity of μ-opioid receptor agonist is limited in the peripheral tissues, and the poison that has so just lowered these type compounds is greatly paid effect.
BKA n/ neutrophilia elastase inhibitor (NEI)
To control whole body and local inflammatory reaction simultaneously as what the front was mentioned, need the multiple pathways of inflammation of blocking-up.Particularly having ability that blocking-up causes the elementary terminal pathway activities factor (as neutrophilia elastoser) relevant with damage with ploidy tissue contusion with the elementary medium (as bradykinin) of keeping inflammatory process, treatment septicemia or other are required serious inflammation that non-enteron aisle treats or the inflammation for the treatment of skin or tooth/periodontal is the key of " single medicine treatment ".
Contain BKA n/ NEI maybe can be decomposed into and acts on two kinds of materials that are present on intracellular two kinds of target spots (bradykinin receptor and neutrophilia elastoser) in conjunction with the active heterogeneous dipolymer compound that can be designed to be kept perfectly.But when wishing that two kinds of active pharmacophoric groups can dissociate, the linking portion that links two active ingredients in the heterogeneous dipolymer can be designed to by as the hydrolysis of blood plasma lytic enzyme.Dissociating or hydrolyzable heterogeneous dipolymer of these kinds is discussed here.BKA n/ inhibitors of cyclooxygenases (COI)
The biological activity of most of bradykinin and the generation of prostaglandin(PG) are closely related.As if for example most of hyperpathia relevant with inflammatory pain depends on damaged tissue and the C-fiber self produces some prostaglandin(PG).To a kind of situation in back, bradykinin and P-material may be the primary stimuluses of these " second messengers " (second messengers).It is that non-bradykinin is dependent that the damaged tissue local produces prostaglandin(PG).The interaction of this proinflammatory medium peptide and prostaglandin(PG) also can take place in other environment, and also can regard a target spot of dual action compounds as.Has BKA n/ COI is in conjunction with the active heterogeneous dipolymer two kinds of pharmacophoric groups that need dissociate in vivo, because epoxidase is an intracellular enzyme and the function bradykinin receptor is limited to outside the serous coat.BKA n/ NK 1Receptor antagonist (NK 1A n)
Bradykinin and P-material known collaborative act on the nervosa agency part that causes and keep inflammation and asthma and multiple pain condition.Under these two kinds of situations, bradykinin is that a kind of very strong (if not the strongest) can stimulate cough sense in mediation peripheral pain and/or the asthma and dyspneic C-fiber to feel the material that imports into.No matter for which kind of primary stimulus, these neurones will discharge the P-material, and it can enlarge and strengthen bradykinin or other stimulator in the activity that stimulates on the sensory nerve ending that is acted on.The initial stimulation of this " list/pair impulsion " (the one/two punch) that is and then amplified by the part has fully been verified and the success or the failure of any single intervention (imtervention) has been had tangible influence.Is target spot by the simplification compound with two aspects of these processes, and the medicine that a kind of dual specificity is provided is possible, and it is better than the monospecific medicine and acts on separately, and easier and cheap than complex therapy.
BKA n/ NK 2Antagonist (NK 2A n)
Bradykinin causes that the ability of the acute contraction of bronchial smooth muscle depends on the same C-fiber that discharges the P-material at least in part and discharges neurokinin A.Neurokinin A is by NK on the bronchial smooth muscle 2Acceptor works.But not only bradykinin can discharge neurokinin A from these neurones, so in conjunction with BKA 2/ NK 2The dual specificity antagonist of antagonistic activity should shrink than the whole better bronchial smooth muscle that improves asthma patient of single medicine.
Usually, heterogeneous dipolymer of the present invention can prepare with the same procedure of describing among the WO 92/17201.This generally relates in the peptide chain appropriate location to BKA nComponent adds and links radicals X, by linking group non-peptide pharmacophoric group is attached to BKA then nOn.Also can be added on the non-peptide pharmacophoric group, then BKA linking group nBe attached on the pharmacophoric group that links base group modification.Although will appreciate that the modification that needs are different, below representative step will be described.
The following example is used for illustrating the present invention, but not limited.
Embodiment 1-4(BKA n/ μ-opioid receptor agonist)
For the present invention is described, four heterogeneous dipolymers of different peptide/opiates (referring to CP-0477, CP-0488, CP-0494 and CP-0499) have been prepared.Wherein three compounds are to use CP0126(DR-R-P-J-G-F-C-DF-L-R) make, the 4th compound is to use CP-02347(DR-R-P-J-G-Thi-C-DTic-Oic-R) preparation.Similarly, link agent with two kinds of different opiates (14-hydroxyl dihydrocodeinone and 14-hydroxyl Hydromorphone) chemistry different and make following heterogeneous dipolymer respectively with two kinds:
The compound compound #The peptide opiate
1 CP-0477 CP-0126 14-hydroxyl dihydrocodeinone
2 CP-0488 CP-0126 14-hydroxyl dihydrocodeinones
3 CP-0494 CP-0126 14-hydroxyl Hydromorphones
4 CP-0499 CP-0347 14-hydroxyl Hydromorphones
The preparation of heterogeneous dipolymer CP-0477, CP-0488, CP-0494 and CP-0499 sees for details following, with reference to accompanying drawing 1:
The preparation of chemical compounds I:
Mix 14-hydroxyl paracodin keto hydrochloride (0.182g, 0.52mmol), acetic acid (0.475ml, 8.3mmol), S-benzyl cysteamine (0.174g, 1.04mmol) and methyl alcohol (5ml) and at room temperature stirred one hour.Add sodium cyanoborohydride (95%, 0.033g, 0.52mmol), and stirring reaction 24 hours at room temperature.The vacuum concentration miscellany.The oil that generates is dissolved in the ethyl acetate, and the ethyl acetate part is used dried over mgso, vacuum-evaporation with the saturated sodium bicarbonate solution washing.Thick product chromatography on silicagel column, with EtOAc, EtOAc-MeOH(9: 1, V/V) and EtOAc-MeOH-Et 3N(9: 1: 0.2, V/V/V) link wash-out, getting chemical compounds I is oily, productive rate 25.0%(59.0mg).
The preparation of compound ii (CP-0477):
(0.059g 0.127mmol) is dissolved in the 2ml anhydrous tetrahydro furan chemical compounds I, is transferred to then in the 100ml three-necked flask of oven dry.Flask and a Dewar (dewar) condenser, nitrogenous source and ammonia inlet tube are connected.Approximately the ammonia condensing of 10ml is in the flask of-78 ℃ of maintenances.The sodium that adds small pieces is used the solid ammonium chloride stopped reaction up to keeping bright blue look after 40 seconds.Reaction mixture rises to room temperature, and ammonia bubbling boiling is overflowed, and adds methyl alcohol (25ml * 3) and vacuum-evaporation.The mercaptan of getting is dissolved in DMF(N in a small amount, dinethylformamide, 2ml) in.Compounds X (about 0.3 equivalent) be dissolved in tris buffer (0.5M, 4.0ml) in, and join in the DMF solution, stirred then 17 hours.Crude mixture is with reverse Vydac C-18HPLC column purification, and is constant with containing 0.1%TFA() water in 15-40%CH 3 CN gradient elution 20 minutes.Retention time is 16.0 minutes.Obtain 26.4mg compound ii white powder after the lyophilize.
Analyze:
Mass spectrum is measured with Finnigan Lasermat mass spectrometer.
Theoretical molecular-1916
Actual measurement molecular weight-1918
Amino acid analysis:
Gly1.02(1), Phe1.92(2 Leu0.97(1 Pro1.01(1 Arg3.14(3)))) and Hyp0.94(1).
The preparation of compound III:
To the 14-hydroxyl paracodin keto hydrochloride that is dissolved in methyl alcohol (10.0ml) (1.0g, 2.84mol) and ammonium acetate (2.2g adds NaC-NBH in mixture 28.4mmol) 3(0.18g, methyl alcohol 2.84mmol) (4.0ml) solution.The solution that forms is transferred to pH7.0 with concentrated hydrochloric acid salt, stirs 17 hours, is acidified to pH1.0 with concentrated hydrochloric acid again.Solvent removed in vacuo, resistates is dissolved in the water.The water layer chloroform extraction is transferred pH9.0 with 10% sodium carbonate solution, with the saturated chloroform extraction of using again of NaCl.Chloroform layer dried over mgso, vacuum-evaporation.Crude product oil purification by silica gel column chromatography, usefulness EtOAc, EtOAc-MeOH(9: 1V/V), EtOAc-MeOH-Et 3N(9: 1: 0.3V/V/V) continuous wash-out.Get the compound III, oily, productive rate are 47.0%(0.42g).
The preparation of compound IV:
With BOC-glycine (0.16g, 0.91mmol), HOBt(0.125g, 0.91mmol) and the 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC) (98.0%, 0.18g 0.91mmol) is dissolved in DMF(2.0ml) in, 0 ℃ was stirred one hour down.To be dissolved in DMF(3.0ml) the amine III (0.24g 0.76mmol) joins in the reaction mixture, and reaction mixture is risen to room temperature, stirs 17 hours.Vacuum is removed DMF, and resultant is dissolved in the ethyl acetate.Ethyl acetate layer with saturated sodium bicarbonate solution and salt water washing, use dried over mgso.Vacuum-evaporation organic layer, crude mixture dodge formula chromatography (flash chromatographed) on silicagel column, use EtOAc-MeOH-Et 3N(9.5: 0.5: 0.3V/V/V) wash-out.Get the oily compound IV, productive rate 82.0%(0.29g).
The preparation of compound V:
In methylene dichloride, use TFA(5.0ml) remove the protecting group BOC of compound IV.Vacuum is removed methylene dichloride, and resistates is used triethylamine (3ml * 3) washing again with methylene dichloride (20ml * 3) washing.With 3-S-benzyl thiohydracrylic acid (0.15g, 0.75mmol), EDC(0.15g, 0.75mmol), HOBt(0.103g, 0.75mmol) and Et 3N(0.35ml 2.48mmol) is dissolved in DMF(5.0ml) in, 0 ℃ was stirred one hour down.To be dissolved in DMF(3.0ml) amine (0.23g 0.62mmol) joins in the reaction mixture.Reaction mixture is warming up to room temperature, restir 17 hours.Vacuum is removed DMF, and resistates is dissolved among the EtOAc, EtOAc layer 10%Na 2CO 3, the salt water washing, dry (use MgSO 4), vacuum-evaporation again.Crude product is purifying on flash silica gel column, uses EtOAc-MeOH-Et 3N(9: 1: 0.3V/V/V) wash-out.Get oily compound V, productive rate 60.0%(0.205g).
The preparation of compound VI (CP-0488)
With the preparation compound ii step with 32.0mg(0.057mmol) compound V deprotection.(0.073g 0.048mmol) reacts in trishydroxymethyl base aminomethane buffer solution for mercaptan that separation obtains and compound X.Crude mixture is carried out purifying with the method for compound ii.The retention time of product is 16.82 minutes.Lyophilize obtains 9.5mg(10%) the white powder VI.
Mass-spectrometric data:
Theoretical molecular 2002
Actual measurement molecular weight 2004
Amino acid analysis:
Gly1.76（2）、Arg3.19（3）、Pro1.06（1）、Leu0.99（1）、Phe2.06（2）、Hyp0.95（1）。
The preparation of compound VII:
With 14-hydroxyl Paramorphan keto hydrochloride (0.56g, 1.06mmol), S-benzyl cysteamine (0.69,4.15mmol), acetic acid (1.52ml, 26.5mmol) and sodium cyanoborohydride (0.11g 1.66mmol) is undertaken by the step of chemical compounds I.The third eluent of purifying crude consist of EtOAc-MeOH-Et 3N(9: 1: 0.3V/V/V).The purified 0.213g oily compound VII that obtains.
The preparation of compound VIII (CP-0494)
(0.063g 0.14mmol) goes protection with VII according to the step of compound ii.(0.335g 0.152mmol) handles in tris buffer the mercaptan that obtains with X.Crude product is with to contain 0.1%TFA(constant) water in 15-70% acetonitrile gradient elution requirement carried out purifying on the inherent anti-phase Vydac C-18HPLC post in following 35 minutes.The retention time of VIII is 15.0 minutes.Cold do 119.0mg(45.0%) white powder compound VI-II.
Mass-spectrometric data:
Theoretical molecular 1902
Actual measurement molecular weight 1904
Amino acid analysis:
Gly0.81（1）、Arg3.12（3）、Pro1.07（1）、Leu0.99（1）、Phe2.04（2）、Hyp0.98（1）。
The preparation of compound IX (CP-0499):
(0.009g 0.02mmol) goes protection by the method for compound ii, and (0.026g 0.016mmol) reacts in tris buffer for mercaptan and XI then with VII.Crude product is purifying on anti-phase Vydac C-18 HPLC, with to contain 0.1%TFA(constant) water in 15-70%CH 3The CN gradient elution, flow velocity 8.0ml/min, 40 minutes time.The retention time of IX is 14.22 minutes.The cold dried white powder IX (6.4mg, 20.0%) that obtains.
Analytical data of mass spectrum:
Theoretical molecular 1957
Actual measurement molecular weight 1958
Experiment in vitro
With rat uterus (BK 2Receptor active) and electricity irritation guinea pig ileum (μ-opiate receptor activity) experiment comment BKA nThe heterogeneous dipolymer of/μ-opioid receptor agonist.These experiments will be known in this area.Gained the results are shown in the table I:
The table I
Compound pA 2-rat uterus IC 50Guinea pig ileum
(nmol)
CP-0126 7.1 non-activities
CP-0347 9.5 non-activities
14-hydroxyl dihydrocodeinone non-activity non-activity
14-hydroxyl Hydromorphone non-activity 21.7
CP-0477 7.9 non-activities
CP-0488 8.2 non-activities
CP-0494??8.4??24.0
CP-0499??8.9??17.0
It should be noted that the heterogeneous dipolymer (CP-0477 and CP-0488) that 14-hydroxyl dihydrocodeinone and 14-hydroxyl dihydrocodeinone form does not all have to show any activity in the active external guinea pig ileum experiment of μ-opioid receptor agonist.This may be that demethylation just has activity because 14-hydroxyl dihydrocodeinone obviously needs in vivo.As a result, can not show activity in this experiment that lacks suitable demethylase.
But more importantly be BKA about these heterogeneous dipolymers nThe data of composition activity and about containing the data of 14-hydroxyl Hydromorphone compound activity.From the table I data as can be seen, used heterodimer has all kept whole BKA nActivity, those utilize 14-hydroxyl Hydromorphone to keep whole μ-opioid receptor agonist activity simultaneously as the compound of opiate.Can obviously find out BKA from these data nThe heterogeneous dipolymer of/μ-opioid receptor agonist can be in vitro system works with their every receptoroid.
Experiment in the body
In order to test these compounds activity in vivo, a kind of inflammation and nervosa pain model have been used.The behavior reaction that this model measurement is showed behind injection 50 μ l Fu Er Malins on the mouse hind leg foot pad.The aggregation of data of these researchs is at Fig. 2, in 3 and 4.Control mice (cavity ring) is the reaction of performance two phase characteristics behind injection Fu Er Malin, and the initial action of a short period of time is wherein arranged, and is a quiet stage then, then is the stage of licking hind leg that continues again.The behavior of licking hind leg means that hind leg is uncomfortable a painful sense.The time of licking is long more, then stings intense pain.
14-hydroxyl Hydromorphone (Fig. 2 A and B) reduces the behavior of licking limb in two stages, but behavior is obviously blunt and cause catalepsy and tangible respiration inhibition when maximal dose (0.9 and 3.0 μ mol/kg).Brad ykinin antagonists CP-0127(effective as selective BK 2Antagonist-Fig. 3 A and B) can reduce in the Fu Er Malin experiment time that two stages lick limb, but dosage will be apparently higher than the used dosage of actual clinical.But CP-0494(Fig. 4 A and B) two stages of not only blocking pain reaction, and used dosage (0.1 μ mol/kg) is starkly lower than 14-hydroxyl Hydromorphone (0.9 μ mol/kg) or CP-0127(12.6 μ mol/kg) dosage when using separately, no less important or what is more important do not have observable anesthetic action in a few hours.These data show just as aforementioned theoretical consideration such as desired, BKA nThe heterogeneous dipolymer of/μ-opioid receptor agonist is better than former pharmacophoric group on pharmacological property.
One skilled in the art will recognize that above-claimed cpd represented one group of compound widely, wherein each component (BKA of heterogeneous dipolymer n, link agent and/or μ-opiate receptor antagonist) all can change, to produce desirable any effect.
Embodiment 5(BKA n/ NEI)
Synthesized a BKA with structure shown in the synthetic route 3 n/ NEI type compound (CP-0502) is used for proving that this compounds can be used as effective part and/or whole body anti-inflammatory drug.This compound is stated compound in the synthetic route 1 (6) as follows from CP-0126 and prototype elastase inhibitor CE-1218().
Synthetic route 1
Figure 931144841_IMG3
Selection is used for the binding group of this heterogeneous dipolymer and links ester bond by the unhinderedly hydrolysis of blood plasma esterase so that make.One skilled in the art will realize that steric restriction that can be by changing ester carbonyl group carbon or make chemical group change acid amides into and link group and modify link group to almost nil so that hydrolysis rate can be from changing fast.Also can make not influenced by potential hydrolysis destructive with completely stable binding residue.
BKA nSynthetic and the analysis of the heterogeneous dipolymer of/NEI
The synthetic of these compounds illustrates that with reference to synthetic route 2 and 3 following detailed synthetic explanation comprises the preparation that suppresses CE-1218 according to the elastoser of synthetic route 1:
Synthetic route 1:
Synthesizing of 4-tertiary butyl phenyl methyl ketone (1)
Have in the 1-L flask to drying and to add CS 2(250ml) and AlCl 3(133.34g 0.56mol), and stirs.This suspension is cooled off with ice bath, drip in 2 hours tert.-butylbenzene (50.00g, 0.37mol) and Acetyl Chloride 98Min. (78.50g, solution 0.41mol) (making temperature be no more than 25 ℃).The stirring at room reaction is spent the night, and is poured into then in the 2L beaker of filled with ice.After the 200ml6NHCl termination reaction, with solution saturated and separation with NaCl.Water layer is with ether washing (2 * 100ml) and merge with organic layer in front.This new organic solution washes (100ml), dry (MgSO with water 4) and evaporation, the oil that obtains distills, and generates 52.1g(79.3%) clear colorless oil 4-tertiary butyl phenyl methyl ketone (bp 0.05Mm70-76 ℃).
1H NMR（CDCl 3）δ1.35（s，9H），2.58（s，3H），7.48（d，J＝8.5Hz，2H），7.91（d，J＝8.5Hz，2H）. 13C NMR（CDCl 3）δ26.42，30.96，34.95，125.36，128.16，134.49，156.64，197.61.
Synthesizing of 4-tert-butyl-phenyl methyl acetate (2)
Be equipped with in the 1L flask of mechanical stirring device and contain Pb(oAc) 4(132.06g is 0.298mol) with 250ml benzene, with nitrogen purging, ice bath cooling.Drip BF in 1 hour introversive this cold slurry 3OEt 2(137.8ml, 1.12mol), 4-tertiary butyl phenyl methyl ketone (50.00g, 0.284mol) solution in 70ml methyl alcohol.Stir that this mixed solution spends the night, water (500ml) termination reaction, again with dilution of 250ml ether and layering.Organic layer water, rare NaHCO 3(carefully) washing, and use MgSO 4Dry.Mixture is after filtration, evaporation and distillation obtain 31.2g(53.4%) clear colorless oil 4-tert-butyl-phenyl methyl acetate (pb 0.04Mm75-80 ℃).
1H NMR（CDCl 3）1.32（s，9H），3.62（s，2H），3.71（s，3H），7.23（d，J＝8.4Hz，2H），7.37（d，J＝8.4Hz，2H）. 13C（CDCl 3）31.33，34.46，40.67，52.04，125.53，128.88，130.91，149.94，172.26.
Synthesizing of 4-tert-butyl-phenyl isopropylformic acid (3)
With 4-tert-butyl-phenyl methyl acetate (30.00g, 0.145mol) and methyl iodide (45.41g, 0.320mol) solution in the anhydrous THF of 125ml was added drop-wise to NaH(8.72g in 30 minutes, 0.363mol) in the slurries in 200mlTHF.After adding, with reaction mixture reflux 1.5 hours.Reactant is cooled to room temperature, with diatomite filtration, concentrated.Resistates is with ether dilution, water washing and use MgSO 4Dry.Generate the oily product of wanting after boiling off solvent.4-tert-butyl-phenyl methyl isobutyrate and contain KOH(10.07g, 0.179mol) 4: 1EtOH/H 2The mixed liquid reflux of O 4 hours.Vacuum boils off E-tOH, raffinate is acidified to pH2 with 2N HCl, leaches solid precipitation.White solid drying (60 ℃, 1mmHg, 24 hours) is obtained target product (23.45g calculates from 4-tert-butyl-phenyl methyl acetate, and productive rate is 73.2%).
1H NMR（CDCl 3）1.34（s，9H），1.62（s，6H），7.37（s，4H），11.4-12.4（brs，1H）. 13C NMR（CDCl 3）26.16，31.30，34.35，45.81，125.31，125.48，140.64，149.66，183.57.
4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carbonyl-tertiary butyl oxygen base-propyl group sulfydryl) phenylester (4) is synthetic:
With 4-tert-butyl-phenyl isopropylformic acid (2.00,0.0091mol) and thionyl chloride (1.62g is 0.0136mol) at 16mlCH 2Cl 2In mixed solution in argon gas, stir and spend the night.Vacuum is removed volatile matter, the solid THF(15ml of formation) dissolving, drip 4-(4 '-hydroxyphenyl in 10 minutes then) sulfydryl butyric acid tertiary butyl ester (2.44g, 0.0091mol), TEA(2.5ml) is at THF(15ml) in solution.Mixture stirs after 3 days and uses Et 2The O dilution is also used 5%NaHCO 3Extract.Organic layer water, salt water washing, dry (MgSO 4).After the evaporation, coloured oil is through separating (HPLC, silicagel column, 70: 30CH 2Cl 2/ hexane is to CH 2Cl 2Linear gradient) obtains target oily product (2.20g, 51.5%).
1H NMR（CDCl 3）δ1.33（s，9H），1.43（s，9H），1.70（s，6H），1.88（tt，J＝7.2Hz，2H），2.35（t，J＝7.2Hz，2H），2.90（t，J＝7.2Hz，2H），6.92（d，J＝8.6Hz，2H），7.32（d，J＝8.6Hz，2H），7.34-7.40（m，4H）. 13C NMR（CDCl 3）δ24.50，26.42，28.08，31.31，33.70，34.11，34.39，46.40，80.40，121.92，125.23，125.46，130.82，133.03，140.86，149.58，149.69，172.21，175.34.
4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-propyl group sulfydryl) phenylester (5) is synthetic:
Stirring down, in 15 minutes trifluoroacetic acid (25ml) joined 4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carbonyl-tert.-butoxy-propyl group sulfydryl) (2.40g is 0.00510mol) at 20mlCH for phenylester 2Cl 2In solution in.Added back 15 minutes, and removed volatile matter, oily matter crystallization (hexane) generates 1.94g(91.8%) the white solid target product.
m.p.86.0-87.0℃， 1H（CDCl 3）1.33（s，9H），1.70（s，6H），1.92（tt，J＝7.0Hz，2H），2.50（t，J＝7.0Hz，2H），2.93（t，J＝7.0Hz，2H），6.93（d，J＝8.7Hz，2H），7.33（d，J＝8.7Hz，2H），7.35-7.39（m，4H）. 13C NMR（CDCl 3）23.89，26.45，31.32，32.34，33.63，34.42，46.42，122.03，125.24，125.48，131.12，132.63，140.85，149.74，175.40，178.65.
4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-sulfonyl propyl base) phenylester (6) is synthetic:
In-50ml flask, add 4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-propyl group sulfydryl) phenylester (1.64g, 0.00396mol), HOAc(25ml) and 25ml 30%H 2O 2Stirring reaction spends the night, and water (50ml) dilutes, and leaches the solid of generation.Dry (12 hours, 1mmHg) after, solid recrystallization (CH 2Cl 2/ hexane) obtain 1.54g(87.1%) the white powder target compound.
mp 107-108.5℃. 1H（CDCl 3）1.33（s，9H），172.（s，6H），2.02（p，J＝7.0Hz，2H），2.52（t，J＝7.0Hz，2H），3.17（t，J＝7.0Hz，2H），7.20（d，J＝8.7Hz，2H），7.36（d，J＝8.7Hz，2H），7.41（d，J＝8.6Hz，2H），7.90（d，J＝8.6Hz，2H）. 13C NMR（CDCl 3）17.88，26.29，31.28，31.79，34.42，46.53，55.04，122.56，125.16，125.61，129.70，135.81，140.22，150.02，155.29，174.73，177.71.
Synthesizing of 6-dimaleoyl imino hexanol (7):
The following synthetic route 2 of synthetic usefulness as the 6-dimaleoyl imino hexanol (7) that links agent illustrates:
Synthetic route 2
Figure 931144841_IMG4
In-100ml flask, add the 6-amino-hexanol (0.76g, 0.0064mol) and the saturated NaHCO of 25ml 3This uniform solution of stirring at room, and adding solid N-methoxycarbonyl maleimide (1.00g, 0.0064mol).Mixture change was soon stirred 1 hour clearly after the adding, and mixture extracts with EtOAc, dry (MgSO 4) and evaporation.The miscellany that generates separates (CH on silicagel column 2Cl 2To EtOAc).Obtain white solid product 0.32g(25.2%), need not to be further purified.
1H（CDCl 3）δ1.25-1.45（m，4H），1.45-1.70（m，4H），3.53（t，J＝7.3Hz，2H），3.63（t，J＝6.0Hz），6.71（s，2H）.
CP-0502's is synthetic:
Compound (6) (CE-1218) forms compound (8) with compound (7) esterification, and then links to generate dipolymer CP-0502 with CP-0126.These are reflected at explanation in the synthetic route 3:
Synthetic route 3
Figure 931144841_IMG5
More specifically with reference to synthetic route 3, and compound (6) (200mg, 0.448mol), triethylamine (0.124ml, 2 equivalents), 6-dimaleoyl imino hexanol (7) (97mg, 1.1i equivalent) be dissolved in the 2ml methylene dichloride.Stir and in this solution, add two (2-oxo-3-Evil alkyl) phospho acid muriates (122mg, 1.0 equivalents) down.The suspension stirring at room that forms 4 hours.Reaction mixture dilutes with methylene dichloride, uses saturated NaHCO 3Washing.The anhydrous MgSO of this organic solution 4Drying, solvent removed in vacuo.Silica gel column chromatography (2 * 18cm post); Acetone/hexanone of 35/65 (Rf=0.4) obtains colorless oil compound (8) as eluent.
Will (50mg 0.08mmol) be dissolved in 10ml and contains among the DMF of 100 μ l diisopropylethylamine through compound (8).Adding 100mg(0.08mmol) CP-0126 stirs reaction down 30 minutes frequently.Reaction mixture injects Vydacl, and " the C-18 reversed-phase column, 15%-90% acetonitrile (containing 0.1%TFA) gradient 10ml/ divided wash-out 30 minutes in the water.With the lyophilize of proper flow part, obtain 52mg(35%) white powder (CP-0502).Laser desorption mass spectrum M/Z=1890(M+H), theoretical value 1890.Automatically the amino acid sequence analysis result has confirmed that amino acid (no change) sequence of peptide is correct.
BKA nThe external activity of the heterogeneous dipolymer of/NEI
The BKA of in-vitro evaluation following compounds nCarry out with standard test method well known in the art (Protocols) with NEI is active.BKA nActive (pA 2) measure NEI(Ki with the rat uterus prepared product Ss) active people's neutrophilia elastoser (HNE) and synthesizing soluble product look substrate methoxyl group succinyl--alanyl-alanyl-prolyl-valyl-o-Nitraniline (MOS-AAPV-pNA) with purifying estimate.Inhibitor in the solution of the NaCl that contains 0.05M sodium phosphate, 0.1M, 0.005% Triton X-100,5%DMSO, pH7.5 with MOS-AAPV-pNA(0.5mM) mixed.Add HNE(10-20nM then).The generation of N-methyl-p-nitroaniline is 25 ℃ with spectrophotometer in wavelength 400-410nm place, temperature and detects down.Use the automatic base of calculation enzyme kinetics of ENZFITTER program parameter then, comprise Ki Ss
Obtain following result:
The table II
Compound pA 2--rat uterus Ki Ss(HNE)
nM
CP-0126 7.1 non-activities
CE-1218 non-activity 10.5
CP-0487 8.4 non-activities
CP-0502??7.5??6.6
Note their Ki values separately, the heterogeneous dipolymer (CP-0502) that the Notes of Key Data in the table II is complete is almost as broad as long with the NEI activity of free sub-NEI fragment monomer (CE-1218).But BKA for complete heterogeneous dipolymer nWith its succinimido hexanol derivative of hydrolysate CP-0487(CP-0126) BKA nThe aspect activity is then different, and wherein complete compound is renderd a service almost than monomers B KA nA low order of magnitude (afullLog) what is interesting is that complete compound shows as the brad ykinin antagonists of irreversible type, and, obviously strengthens the uterotonic antagonistic activity of bradykinin inductive when longer in the incubation time.Such receptor acting can not be used standard pA 2Analyze well and measure, so BKA between observed CP-0487 and the CP-0502 nActivity difference may be more obvious than actual.Regardless of these numbers molecular pharmacology mechanism behind, clearly can be with BKA nBe incorporated in the single molecule with the NEI activity.
The above-mentioned Notes of Key Data considers that effect behavior that hydrolysis in the body of complete compound can change both is to strengthen former compound activity in vivo.The animal model of unfortunately not set up can be used for measuring BKA in the body nActive with the combination of NEI.Therefore, in order to measure complete heterogeneous dipolymer hydrolysis potentiality in vivo, used a kind of external " substituting " system, wherein heterogeneous dipolymer (CP-0502) is with the human plasma incubation, then the metabolite that produces with the reversed-phase HPLC analysis.
CP-0502 is added in the human plasma of newly getting, and at 37 ℃ of incubation different times.Under the time of these settings, use acidifying (0.1NHCl) acetonitrile treatment sample with the precipitation plasma proteins.The sample aliquot of supernatant liquor (75 μ l) is analyzed under 24% to the 80% acetonitrile gradient elution requirement in 0.1%TFA on Vydac C-18 reversed-phase HPLC post.Elutriant detects under 214nm.
The reversed-phase HPLC color atlas of Fig. 5 a and this analysis of b representative explanation.As can be seen, parent compound be it seems the Arg that takes off that is hydrolyzed to monomer CP-0487 that the succinimido hexanol modifies and it soon from these color atlass 9Derivative (blood plasma carboxypeptidase will be from complete heterogeneous dipolymer CP-0502 and CP-0487 the terminal arginine residues of cracking).The apparent T1/2 of this hydrolysis reaction is approximately 113 minutes.The NEI component of heterogeneous dipolymer is an active ester, and is hydrolyzed simultaneously.But complete NEI component and its hydrolysate are covered from the peak of blood plasma in the middle part of being schemed, and can't make its visibleization with this system.Because NEI has same activity as heterogeneous dipolymer component with as monomer, its active influence is less than BKA so heterogeneous dipolymer is decomposed into two parts nComponent.
Those skilled in the art will recognize that pulsating solid of " binding " ester and electronic environment can influence the hydrolysis rate of heterogeneous dipolymer, used here chemical group is only for can be used to regulate the heterogeneous dipolymer example of chemical group of speed (or not dissociating) that dissociates.
Embodiment 6(BKA n/ COI)
BKA nSynthetic and the analysis of the heterogeneous dipolymer of/COI
Synthesized representative BKA according to following synthetic route 4 and 5 nThe heterogeneous dipolymer of/COI (CP-0460).
Synthetic route 4
Figure 931144841_IMG6
The 1-(4-chlorobenzene formacyl)-5-methoxyl group-2-methyl-3-indolyl acetic acid 6-dimaleoyl imino polyhexamethylene (9) synthetic:
In-100ml flask, add INDOMETHACIN (1.90g, 0.00532mol), 25mlCH 2Cl 2And DCC(0.55g, 0.00266mol).After 2 hours, miscellany is filtered, use 15mlCH 2Cl 2Washing dicyclohexylurea (DCU), (0.50g 0.00253mol), adds anhydrous Na again to add solid 6-dimaleoyl imino hexanol in this new soln 2CO 3(0.32g, 0.00304mol).After 4 days, miscellany is filtered, use Et 25%NaH-CO is used in the O dilution 3, H 2O washing and dry (MgSO 4).The yellow oil HPLC(silica gel that generates; CH 2Cl 2To 80: 20CH 2Cl 2/ EtOAc linear gradient, 60 minutes) purifying, obtain 0.79g(58.0%) the yellow oily target product. 1HNMR
（CDCl 3）1.20-1.35（m，4H），2.38（s，3H），3.47（t，J＝7.3Hz，2H），3.66（s，2H），3.83（s，3H），4.08（t，J＝6.6Hz，2H），6.66（J＝9.0Hz，J＝2.5Hz，1H），6.68（s，1H），6.87（d，J＝9.0Hz，1H），6.96（d，J＝2.5Hz，1H），7.47（d，J＝8.5Hz，2H），7.66（d，J＝8.5Hz，2H）.
Compound (9) links generation CP-0460 with CP-0126 and illustrates in synthetic route 5 and be described below:
Synthetic route 5
Figure 931144841_IMG7
CP-0126(100mg, 0.08mmol) with compound (9) (0.12mmol, 1.5 equivalents) 2ml95%DMF/5% contain stir frequently in the bicarbonate of ammonia of 50 μ l diisopropylethylamine under reaction 30 minutes.Reaction mixture is injected Vydac 1 " the C-18 reversed-phase column is at flow velocity 10ml/ branch, in 20 minutes from 15% acetonitrile/0.1%TFA to 40% acetonitrile/the 0.1%TFA condition of gradient elution under purifying.Generate 64mg(45% after the lyophilize of proper flow part) colourless powder (CP-0460).The laser dissociation mass spectrum: M/Z=1802(M+H), theoretical value 1802.
As previously mentioned, COI works needs and BKA nDissociate so that it is penetrated in the cell.Therefore, in order to estimate BKA nThe function activity of the heterogeneous dipolymer of/COI acts on CP-0460 on the induced lung essence bar, uses the arachidonic acid excitement then.This organizes knownly not only has non-specificity esterase activity but also tetraenoic acid can be changed into thromboxane (by the epoxidase dependent pathway), and the latter finally causes observed smooth muscle contraction in this experiment.
Use this system, find that the log10 dose of thromboxane and CP-0460 is respectively 0.988+/-0.425 and 1.029-/-0.042 than displacement, the private INDOMETHACIN of instruction book and CP-0460 suppress the contraction that exogenous arachidonic acid causes and render a service identical.BKA nThe log10 dose of itself does not influence this system than shift value.These data presentation BKA nThe COI component of the heterogeneous dipolymer of/COI has at the same time in ester hydrolysis and the active tissue of epoxidase the function activity is arranged.
Also tested the BKA of complete CP-0460 with the experiment of standard rat uterus nActivity, the pA of discovery CP-0460 2Be approximately 7.8.In addition, CP-0460(is similar to CP-0502) do not show as the uterotonic antagonist of striving property unexpectedly that typical bradykinin brings out, and be a kind of " false non-striving property unexpectedly " antagonist (when particularly concentration is higher).It is active own that this atypia behavior does not relate to COI, because INDOMETHACIN concentration height is to all not influences of this experiment.
In any case explain viewed data, as other two compounds of this paper explanation, those skilled in the art can be familiar with and can prepare BKA important on the pharmacology with various suitable binding segment nThe heterogeneous dipolymer of/COI has the heterogeneous dipolymer of hydrolyzable ester so that the carboxylic group of free hydroxyl and COI component (general characteristics of many COI) forms.These compounds can be used for treating various inflammation or pain condition and treatment uterine smooth muscle malfunction.
The present invention has exemplified the example of the non-peptide Y of above-mentioned application component, and this component also can be made up of as a kind of peptide that exemplifies in aforementioned WO 92/172001 whole or part, comprises the heterogeneous dipolymer that this literary composition is described.
Dipolymer of the present invention can be used with the conventional medicament composition forms of being made up of active constituent and pharmaceutical carrier.These compositions are applicable to local, oral, aerosol, intramuscular, subcutaneous or intravenously administrable.The content of active constituent can be according to purposes and route of administration for example changing between 0.001 and 90.0%, although available more or less active ingredient in these compositions.Dosage commonly used will change according to concrete purposes and administering mode to a great extent.But general effective dose is between per kilogram of body weight 0.1 to the 1000mg order of magnitude.
Scope of the present invention such as following claim define.

Claims (35)

1, the heterogeneous dipolymer of following formula:
BKA wherein nBe the brad ykinin antagonists peptide; Y one is different from BKA nPharmacophoric group; X is that chemistry links BKA nBinding fragment with the Y component.
2, according to the heterogeneous dipolymer of claim 1, wherein Y is the active non-peptide pharmacophoric group with non-kassinin kinin part of antagonism inflammatory process.
3, according to the heterogeneous dipolymer of claim 1, wherein Y is the active peptide class pharmacophoric group with non-kassinin kinin part of antagonism inflammatory process.
4, according to the heterogeneous dipolymer of claim 1, wherein Y is μ-opioid receptor agonist.
5, according to the heterogeneous dipolymer of claim 1, wherein Y is the neutrophilia elastase inhibitor.
6, according to the heterogeneous dipolymer of claim 1, wherein Y is an inhibitors of cyclooxygenases.
7, according to the heterogeneous dipolymer of claim 1, wherein Y is NK 1Receptor antagonist or NK 2Receptor antagonist.
8, according to the heterogeneous dipolymer of claim 1, wherein X is hydrolyzable.
9, according to the heterogeneous dipolymer of claim 1, wherein X is a non-hydrolysable.
10, according to the heterogeneous dipolymer of claim 1, wherein X is by mixing at BKA nIn amino acid or amino acid analogue form.
11, according to the heterogeneous dipolymer of claim 1, wherein X is made up of the binding group based on maleimide/succinimide.
12, according to the heterogeneous dipolymer of claim 1, wherein X is made up of double amber imide base alkane.
13, according to the heterogeneous dipolymer of claim 1, wherein X is by BKA nThe S atom of peptide sulfydryl is formed.
14, according to the heterogeneous dipolymer of claim 1, wherein Y is selected from 14-hydroxyl dihydrocodeinone or 14-hydroxyl Hydromorphone.
15, according to the heterogeneous dipolymer of claim 1, wherein Y is an INDOMETHACIN.
16, according to the heterogeneous dipolymer of claim 1, wherein Y is that neutrophilia elastase inhibitor and X comprise and pass through BKA nThe sulphur atom of sulfydryl is attached at BKA on the chain nOn the succinimide group.
17, according to the heterogeneous dipolymer of claim 16, wherein the neutrophilia elastase inhibitor is 4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-sulfonyl propyl base) phenylester.
18, comprise in order to be attached to BKA nOn the peptide and the pharmacophoric group of the binding group that adds.
19,, wherein link group and form by the amino acid that amine, carboxylic acid, hydroxyl, sulfydryl or maleimide base group maybe can be incorporated in the peptide according to the pharmacophoric group of claim 18.
20, be included as the BKA of the binding group that links non-peptide pharmacophoric group and add nPeptide.
21, according to the BKA of claim 20 nPeptide, wherein linking group is that terminal alkane chain, amine or hydroxy-acid group formed by sulfydryl, maleimide base group or with the hydroxyl.
22, according to the pharmacophoric group of claim 18, it is μ-opioid receptor agonist.
23, according to the pharmacophoric group of claim 18, it is the neutrophilia elastase inhibitor.
24, according to the pharmacophoric group of claim 18, it is an inhibitors of cyclooxygenases.
25, according to the pharmacophoric group of claim 18, it is NK 1Receptor antagonist or NK 2Receptor antagonist.
26, according to the heterogeneous dipolymer of claim 1, BKA wherein nBe DR-R-P-J-G-F-C-DF-L-R or DR-R-P-J-G-Thi-G-DTic-Oic-R.
27, according to the heterogeneous dipolymer of claim 26, wherein another pharmacophoric group is μ-opioid receptor agonist.
28, according to the heterogeneous dipolymer of claim 26, wherein another pharmacophoric group is the neutrophilia elastase inhibitor.
29, according to the heterogeneous dipolymer of claim 26, wherein another pharmacophoric group is an inhibitors of cyclooxygenases.
30, according to the heterogeneous dipolymer of claim 26, wherein another pharmacophoric group is NK 1Antagonist or NK 2Antagonist.
31, according to the heterogeneous dipolymer of claim 28, wherein pharmacophoric group is 4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-propyl group-alkylsulfonyl) phenylester.
32, according to the heterogeneous dipolymer of claim 31, wherein X is by being attached to BKA with-S-atom nTwo amber imido grpup groups on the component are formed.
33, phenylester compound 4-tert-butyl-phenyl isopropylformic acid 4-(3 '-carboxyl-sulfonyl propyl base).
34, the pharmaceutical composition of forming by the heterogeneous dipolymer and the pharmaceutical carrier of claim 1.
35, in method, use the improvement that heterogeneous dipolymer constituted of claim 1 with brad ykinin antagonists treatment pain or inflammation.
CN93114484A 1992-11-10 1993-11-10 Brad ykinin antagonists Pending CN1094058A (en)

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Families Citing this family (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995003308A1 (en) * 1993-07-23 1995-02-02 Toray Industries, Inc. Morphinan derivative and medicinal use
AU6044496A (en) * 1995-06-05 1996-12-24 Cortech, Inc. Compounds having bradykinin antagonistic activity and mu-opi oid agonistic activity
CA2221865C (en) * 1995-06-09 2007-12-11 The Regents Of The University Of Michigan Bradykinin analogs as selective thrombin inhibitors
FR2739553B1 (en) * 1995-10-06 1998-01-02 Oreal USE OF BRADYKININE ANTAGONISTS TO STIMULATE OR INDUCE HAIR GROWTH AND / OR STOP THE HAIR LOSS
DE19612067A1 (en) * 1996-03-27 1997-10-02 Hoechst Ag Use of bradykinin antagonists for the manufacture of medicaments for the treatment of chronic fibrogenetic liver diseases and acute liver diseases
DE19642289A1 (en) 1996-10-14 1998-04-16 Hoechst Ag Use of bradykinin antagonists for the manufacture of medicaments for the treatment and prevention of Alzheimer's disease
US6982249B1 (en) 1997-04-23 2006-01-03 The Regents Of The University Of Michigan Bradykinin analogs as selective inhibitors of cell activation
US6359111B1 (en) * 1998-05-28 2002-03-19 Neorx Corporation Opioid receptor targeting
US6716452B1 (en) 2000-08-22 2004-04-06 New River Pharmaceuticals Inc. Active agent delivery systems and methods for protecting and administering active agents
US7060708B2 (en) 1999-03-10 2006-06-13 New River Pharmaceuticals Inc. Active agent delivery systems and methods for protecting and administering active agents
WO2003020200A2 (en) 2000-11-16 2003-03-13 New River Pharmaceuticals Inc. A novel pharmaceutical compound and methods of making and using same
EP2080511A3 (en) * 2000-08-22 2009-09-30 Shire LLC Active agent delivery systems and methods for protecting and administering active agents
AU2001297565B2 (en) * 2000-11-16 2008-06-05 Takeda Pharmaceutical Company Limited A novel pharmaceutical compound and methods of making and using same
ES2373641T3 (en) * 2000-11-16 2012-02-07 Shire Llc A NEW PHARMACEUTICAL COMPOUND AND METHODS TO MANUFACTURE AND USE.
US7375082B2 (en) 2002-02-22 2008-05-20 Shire Llc Abuse-resistant hydrocodone compounds
US7338939B2 (en) 2003-09-30 2008-03-04 New River Pharmaceuticals Inc. Abuse-resistant hydrocodone compounds
US7169752B2 (en) 2003-09-30 2007-01-30 New River Pharmaceuticals Inc. Compounds and compositions for prevention of overdose of oxycodone
US7700561B2 (en) 2002-02-22 2010-04-20 Shire Llc Abuse-resistant amphetamine prodrugs
IL163667A0 (en) 2002-02-22 2005-12-18 New River Pharmaceuticals Inc Novel sustained release pharmaceutical compounds to preventabuse of controlled substances
AR040956A1 (en) 2002-07-31 2005-04-27 Schering Ag NEW CONJUGATES OF EFFECTORS, PROCEDURES FOR THEIR PREPARATION AND PHARMACEUTICAL USE
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SG147485A1 (en) 2003-09-30 2008-11-28 Shire Llc Pharmaceutical compositions for prevention of overdose or abuse
GB0624880D0 (en) * 2006-12-14 2007-01-24 Johnson Matthey Plc Improved method for making analgesics

Family Cites Families (4)

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Publication number Priority date Publication date Assignee Title
US4894443A (en) * 1984-02-08 1990-01-16 Cetus Corporation Toxin conjugates
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CA2106677A1 (en) * 1991-04-01 1992-10-02 John C. Cheronis Bradykinin antagonists
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