WO1993008823A1 - Composes de guanidinyle et composes apparentes modulant l'adhesion cellulaire - Google Patents

Composes de guanidinyle et composes apparentes modulant l'adhesion cellulaire Download PDF

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WO1993008823A1
WO1993008823A1 PCT/US1991/006469 US9106469W WO9308823A1 WO 1993008823 A1 WO1993008823 A1 WO 1993008823A1 US 9106469 W US9106469 W US 9106469W WO 9308823 A1 WO9308823 A1 WO 9308823A1
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compound
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PCT/US1991/006469
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Thomas C. Mckenzie
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Tanabe Seiyaku Co., Ltd.
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06052—Val-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/04—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms
    • C07C275/20—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an unsaturated carbon skeleton
    • C07C275/24—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to acyclic carbon atoms of an unsaturated carbon skeleton containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0205—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0207—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)4-C(=0), e.g. 'isosters', replacing two amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/021—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)n-C(=0)-, n being 5 or 6; for n > 6, classification in C07K5/06 - C07K5/10, according to the moiety having normal peptide bonds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06104—Dipeptides with the first amino acid being acidic
    • C07K5/06113—Asp- or Asn-amino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel guanidinyl and related compounds which are characterized by cell adhesion modulation activity.
  • the compounds have application to the study and treatment of disease conditions mediated by cell adhesion.
  • the compounds have application to the study, diagnosis, treatment or prevention of diseases and conditions such as, for example, cardiovascular disease, harmful platelet aggregation, neoplastic disease including metastasis of neoplastic growth, wound healing, inflammation and autoimmune disease or other diseases or conditions involving cell adhesion.
  • the extracellular matrix is that material which surrounds the muscle and is the major component of connective tissue of all mammals.
  • the extracellular matrix provides for structural integrity, and promotes cell migration and cellular differentiation. As part of these functions, the extracellular matrix has been shown to support adhesion for various types of cells in vitro. Molecules such as the collagen, fibronectin, vitronectin, laminin, von Willebrand factor, thrombospondin, bone sialoprotein, fibrinogen, and tenacin have been found to possess this property of mediating cell adhesion.
  • the above cell-adhesive molecules have been found to exhibit a structural similarity in their respective binding sites, each of which contains the amino acid sequence arginine-glycine-aspartic acid, or RGD using conventional single letter nomenclature.
  • the cell-binding site in fibronectin has been reproduced synthetically.
  • the cellular receptor site for fibronectin has been identified for various cells.
  • cellular receptors that recognize RGD-containing sequences in other extracellular matrix proteins e.g., the vitronectin receptor
  • Such cellular receptors responsive to RGD-containing proteinaceous compounds, have been characterized.
  • the complete, primary structure of the fibronectin receptor has been deduced from cDNA and physical properties have been determined.
  • the protein exists at the cell surface as a heterodimeric complex (although the larger polypeptide is enzymatically processed) having both polypeptide chains inserted into the membrane. Each chain extends 30-40 residues into the cytoplasmic space, and at least one of the cytoplasmic peptides appears to interact with the cytoskeieton.
  • the ⁇ subunit is somewhat smaller and conformationally compact due to numerous intrachain disulfide bonds.
  • the cytoplasmic domain of the ⁇ subunit contains a potentially phosphorylated tyrosine. Hirst et al., PNAS-USA, 1986, 83, 6470; Tamku ⁇ et al., Cell, 1986, 46, 271-282.
  • RGD-directed receptors as well as other "orphan" receptors the Iigand for which is unknown, have also been characterized.
  • This putative RGD commonality of the Iigand matrix proteins has revealed a superfamily of cell surface receptor proteins that share a high degree of structural similarity and probably also functional similarity.
  • the members of this superfamily of cell surface proteins collectively are known as the integrins.
  • the integrins can be grouped on the basis of the identity of their ⁇ subunit.
  • the ⁇ subunit as disclosed above for the fibronectin receptor, is compact due to a high degree of cross-linking.
  • the first group of integrins includes the very late activation antigen (VLA) proteins, which themselves include the fibronectin receptor (VLA-5), the collagen receptor (VLA-2), and the laminin receptor.
  • the second group includes the lymphocyte associated antigen-l (LFA-I), macrophage antigen-l (MAC-1), and p150,95.
  • the third group includes the vitronectin receptor, and platelet glycoprotein gpllb/llla. Hynes, Cell, 1987, 48, 549; Hemler, Immunol. Today, 1988, 9, 109; Springer et al., Annu. Rev. Immunol., 1987, 5, 223; Kishimoto et al., Leukocyte Adhesion Molecules, T.A. Springer, D.C. Anderson, A.S. Rosenthal, and R. Rothlein, Ed., Springer-Verlag: New York; 1989, pp. 7-43.
  • the RGD-directed receptor present on platelets that binds fibronectin, vitronectin, fibrinogen, and von Willebrand factor has also been purified.
  • This receptor is the gpllb/IIIa protein complex.
  • This receptor is thus not specific to one extracellular matrix protein, as are the above fibronectin and vitronectin receptors, ft has been proposed that this lack of specificity is correlated to the lack of conformationa! specificity in the ligands.
  • Other work has suggested that specificity can be achieved with relatively short, conformationally restricted synthetic peptides containing the RGD sequence.
  • the receptor affinity for its peptide Iigand may be altered as the stereoconformation, or three-dimensional shape, of the peptide is restricted, typically by cyclization. Pierschbacher and Ruoslahti, PCT International Publication WO 89/05150 (1989).
  • a limited number of compounds containing sequences of natural amino acids or derivatives other than RGD may also possess the capability for affecting cell adhesion. These non-RGD-containing peptides are not well characterized. See,
  • U.S. Patent No.4,879,313 to Tjoeng, ef al. reports the utility as platelet aggregation inhibitors of certain peptide mimetic compounds containing, in addition to a guanidinyl group at one terminus and an internal aspartic acid residue, an aromatic structure (phenyl, biphenyl, naphthyl, pyridyl or thienyl groups, and certain methoxy-substituted forms thereof) at another defined position in the compound.
  • Related structures containing an internal glycine residue are reported in U.S. Patent No. 4,857,508 to Adams, ef al.
  • the present invention relates to compounds having activity as cell adhesion modulators.
  • the compounds are characterized by the presence of a guanidinyl or related group (e.g., substituted guanidinyl, urea or thiourea group) linked through a hydrocarbon moiety, and optionally also through an amino acid residue, to a carboxyl- or carboxyl-derivative-containing portion comprising an aspartic acid- related residue and, optionally, an additional amino acid residue or analog thereof.
  • the carboxyl or carboxyl-derivative portion of the compounds may have either a normal- or a reverse-orientation peptide bond structure, and (especially in the latter case) may contain one or more D-enantiomer residues.
  • the compounds in one aspect, sufficiently mimic extracellular matrix ligands or other cell adhesion ligands so as to bind to cell surface receptors.
  • Such receptors include integrin receptors in general, including the fibronectin, collagen, laminin, LFA-1 , MAC-1, p150,95, vitronectin and gpllb/llla receptors.
  • the novel compounds have been found to modulate cell adhesion by competing, for example, with RGD-containing ligands and by binding to RGD-directed receptors on cell surfaces.
  • Such cell adhesion ligands including (but not limited to) fibronectin, are sufficiently inhibited from binding to the cell's receptor as to prevent or reduce cell adhesion.
  • Other uses include enhancing cell adhesion by using the compounds to attach cells to a surface, or by other promotion of cell adhesion.
  • the useful compounds herein described function as cell-adhesion modulators.
  • One object of the present invention is to provide novel compounds which act to modulate cell adhesion. Another object of the present invention is to provide novel guanidinyl and related compounds which are capable of binding with a cellular receptor.
  • Another object of the present invention is to provide novel guanidinyl and related compounds which contain one or more "reverse" amino acid residues and which are capable of binding to a cellular receptor.
  • Another object of the present invention is to provide a novel method for modulating cell adhesion using novel compounds.
  • Another object of the present invention is to provide novel compounds, formulations, and methods which may be used in the study, diagnosis, treatment or prevention of diseases and conditions which relate to cell adhesion, including but not limited to rheumatoid arthritis, asthma, allergies, adult respiratory distress syndrome (ARDS), cardiovascular disease, thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, neoplastic disease including metastasis of neoplastic growth, wound healing, Type I diabetes, inflammatory conditions including ophthalmic inflammatory conditions and inflammatory bowel disease (e.g, ulcerative colitis and regional enteritis), and autoimmune diseases.
  • diseases and conditions which relate to cell adhesion, including but not limited to rheumatoid arthritis, asthma, allergies, adult respiratory distress syndrome (ARDS), cardiovascular disease, thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, neoplastic disease including metastasis of neoplastic growth, wound healing, Type I diabetes,
  • Another object is to provide derivative compounds, such as, but not limited to, antibodies and anti-idiotype antibodies to the compounds disclosed and claimed in order to study, diagnose, treat or prevent diseases and conditions which relate to cell adhesion, including but not limited to rheumatoid arthritis, asthma, allergies, adult respiratory distress syndrome (ARDS), cardiovascular disease, thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, neoplastic disease including metastasis of neoplastic growth, wound healing, Type I diabetes, inflammatory conditions and autoimmune diseases.
  • rheumatoid arthritis asthma
  • allergies adult respiratory distress syndrome
  • cardiovascular disease thrombosis or harmful platelet aggregation
  • reocclusion following thrombolysis neoplastic disease including metastasis of neoplastic growth, wound healing, Type I diabetes, inflammatory conditions and autoimmune diseases.
  • FIGURE 1 is a graphical representation showing cell adhesion inhibition by a representative guanidinyl compound of the invention, 8-guanidinyloctanoy-(O-phenylmethyl)-aspartyl-(O-phenylmethyl)serine phenylmethyl ester.
  • the compounds of the present invention are those having the property of modulating cell adhesion.
  • ARDS adult respiratory distress syndrome
  • the attachment of inappropriate cells to the lung lining induces an inflammatory response.
  • Preliminary in vitro results show that such detrimental attachment, in which the leukocyte adheres to endothelial cells or the lung extracellular matrix, is mediated by RGD-containing protein and RGD-recognizing receptors on the leukocytes.
  • peptides or other compounds with a binding affinity to RGD receptors are desirable as competitive antagonists and should be useful in treating ARDS and asthma. Such compounds are disclosed herein.
  • a peptide or other compound with suitable affinity for RGD receptors, such as disclosed herein, should likewise have anti-metastasis utility. Harmful blood clotting is also caused by inappropriate cell adhesion, particularly cell adhesion to the extracellular matrix.
  • the attachment, spreading and aggregation of platelets on extracellular matrices are central events in thrombus formation. These events can be regulated by the family of platelet adhesive giycoproteins, fibrinogen, fibronectin, and von Willebrand factor.
  • Fibrinogen functions as a cofactor for platelet aggregation, while fibronectin supports platelet attachment and spreading reactions. Von Willebrand factor is important in platelet attachment to and spreading on subendothelial matrices. Plow ef al., PNAS-USA, 1985, 82, 8057. A peptide or other compound, such as these herein, which would function as an antagonist and bind to cell receptors which recognize the matrix glycoprotein RGD site would be beneficial as a thrombolytic.
  • a peptide or other compound with suitable affinity for RGD receptors attached for example to a suitably positioned matrix or surface, may be able to promote beneficial cell adhesion and resultant wound healing by binding cells with the appropriate RGD-recognizing receptor.
  • such peptides or other compounds coating the prosthesis would provide a means for covering the prosthesis with a surface of cells. This cell surface would provide a surface compatible with the organism, and thus minimize rejection that might otherwise occur due to stimulation of the immune system by the prosthesis itself.
  • the compounds of the present invention are believed to be useful in this cell adhesion modulation application as well.
  • the cell adhesion modulation compounds of the present invention are represented in part herein by amino acid residues wherein the individual amino acids are represented by their standard three-letter abbreviations as follows:
  • the first group of compounds of this invention are of the formula:
  • X is selected from NR, S and O;
  • Y is an unsubstituted or substituted, linear or branched linking moiety selected from saturated and unsaturated hydrocarbon groups containing from 1 to about 15 linking atoms, and optionally containing one or more heteroatoms and/or one or more cyclic structures;
  • Z is optional and, where present, is selected from Gly (most preferably), and from Ala, Sar and ⁇ Ala;
  • R 1 and R 2 are independently substitue ⁇ ts selected from -OR (including hydroxyl), -NR 2 (including -NH 2 and -NHR), -NHNH 2 and -SR, and where one of R 1 and R 2 may additionally be selected from
  • R 3 is a substituent selected from -OR (including hydroxyl), -NR 2 ( ⁇ nduding -NH 2 and -NHR), -NHNH 2 and -SR, and
  • R 4 is a substituent selected from groups of the form -R and acyl groups
  • n and (where present) m are independently 1, 2 or 3;
  • each R is individually a pharmaceutically suitable substituent group, preferably one selected from hydrogen, from linear and branched, unsubstituted and substituted C 1 -C 8 lower alkyls, C 2 -C 8 alkenyls, C 2 -C 8 alkynyls, C 3 -C 14 aryls, C 7 -C 14 alkaryls, C 7 -C 14 aralkyls and C 3 -C 14 cycloalkyls, from heteroatomic groups and, in the case of -NR 2 , from cyclized groups forming (in attachment with the nitrogen atom) a 5-8 membered heterocyclic ring optionally containing oxygen, nitrogen or sulfur as a further ring heteroatom.
  • substituent group preferably one selected from hydrogen, from linear and branched, unsubstituted and substituted C 1 -C 8 lower alkyls, C 2 -C 8 alkenyls, C 2 -C 8 alkynyls, C 3 -C
  • the compounds of the invention further include pharmaceutically acceptable baseor acid-addition salts of the compositions of Formula I.
  • the pharmaceutical compositions of the invention include such compounds (including salts thereof) formulated with a pharmaceutically acceptable excipient.
  • Methyl, ethyl, isopropyl, fe/f-butyl and benzyl are examples of such preferred hydrocarbon substituents, with benzyl (-CH 2 (C 6 H 5 )) being a particularly preferred aralkyl substituent.
  • Homoatomic polycyclic structures such as naphthalene, decalin, anthracene or phenanthrene may also be employed, although single-ring structures are presently preferred for cyclic substituents.
  • Each R group in the compound may be independently selected, and need not be the same in each position in the compound.
  • the cyclic structure may contain one or more heteroatoms selected from N, O and S, and may be mono- or polycyclic.
  • the cyclic structure can be saturated, as in morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, imidazolidinyl, and other structures, or unsaturated or aromatic, as in imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyrrolyl, pyrrolinyl, pyridazinyl, pyrrodiazolyl, isothiazolyl, thienyl, thiazinyl, isoxazolyl, furaza ⁇ yl and other structures.
  • Potycydic structures such as indolyl, quinoiyl, quinazolinyl, phenoxazinyl, phenazinyl, phenothiazinyl, benzo[b]thienyl, phe ⁇ anthrolinyl or others may be employed. Attachment of such cyclic R groups with the remainder of the compound may occur through a carbon or (provided that a point of bonding is present on a heteroatom) through a heteroatom within the heterocyclic group, or attachment may be achieved through, for example, an intermediate alkylene moiety which links the cyclic group with the remainder of the compound. Such cydic R groups may also be substituted with pharmaceutically suitable substituents as is now discussed.
  • substituents include hydroxyl, amino, lower (C 1 -C 8 ) alkoxyl and alkyl, and, in the case particularly of aromatic R groups, the foregoing substituents as well as nitro and halogen (especially chloro and bromo) moieties.
  • substituents on R may be used, for example, to after bioactivity, solubility and/or biodistribution characteristics of the subject compounds.
  • R includes an aryl group, substituents occurring on the meta and/or para positions (i.e., 3'-and/or 4'-positions) are most preferred.
  • Preferred alkaryl forms of R thus include (3'-methyl)phenyl and (4'-methyl)phenyl groups. Multiply-substituted aryls, particularly where the substituent is small as in methyl or halo, are also useful.
  • the preferred R substituents in this position are as discussed above, with hydrogen being most highly preferred (so as to yleld an unsubstituted guanidinyl moiety).
  • Lower alkyl and single-ring aryl (including single-ring aralkyl and alkaryl) moieties are also preferred.
  • R 1 , R 2 , R 3 and R 4 those of the form -OR, with choices of R therein ylelding hydroxyl substituents (i.e., R is hydrogen), as well as those ylelding unsubstituted and substituted lower alkoxy and single-ring aryloxy, and unsubstituted and singly-substituted lower alkylamino substituents, are preferred.
  • R may be the same or different in each position in R 1 ,
  • R 2 , R 3 and R 4 where such a substituent ylelds a structure of the form -OR other than hydroxyl (i.e., R is other than hydrogen), structures of the form -OCH 3 , -OCH 2 (C 6 H 5 ) and -O(C 6 H 5 ) (i.e., -R is -CH 3 , -CH 2 (C 6 H 5 ) or -(C 6 H 5 ) are especially preferred and are believed to protect the resultant compound from metabolism in the in vivo environment, and may also tend to enhance the ability of the compound to interact with receptor sites on cells or other structures in the body, thereby increasing the activity of the compound.
  • R 1 , R 2 and/or R 3 are of the form -NR 2
  • amidating substituents such as those of the form -NH 2 , -NHCH 3 and -NHCH 2 CH 3 (i.e., each -R is hydrogen, or one -R is hydrogen and the other is -CH 3 or -CH 2 CH 3 )
  • amidatinq substituents wherein the nitrogen is itself substituted with one or two aromatic groups i.e., one or two R groups in -NR 2 is of an aryl, alkaryl or aralkyl form as defined above
  • R 1 , R 2 , and R 3 bulky esterifylng groups, such as aromatic esterifylng groups, of the form -OR are particularly preferred, and the benzyloxy (-OCH 2 (C 6 H 5 )) group is particularly preferred for each of R 1 , R 2 and R 3 .
  • R 4 it is espedally preferred that R therein be an aromatic group, particularly benzyl or benzoyl.
  • substituents on such aromatic groups including hydroxyl, amino, lower aikoxyl and alkyl substituents, and espedally electron-withdrawing substituents such as halogen and nitro groups, are useful in one or more positions on the aromatic structure.
  • substituents aromatic and otherwise in these positions include those wherein R is p-chlorobenzyl, benzyloxymethyl, trityl (triphenylmethyl), t-butyl, dnnamyl, other substituted benzyl or substituted methyl, alkyl, cycloalkyl, phenyldimethylsilyl, t-butyldimethylsilyl and triisopropylsilyl.
  • R is p-chlorobenzyl, benzyloxymethyl, trityl (triphenylmethyl), t-butyl, dnnamyl, other substituted benzyl or substituted methyl, alkyl, cycloalkyl, phenyldimethylsilyl, t-butyldimethylsilyl and triisopropylsilyl.
  • N-morpholinyf heterocyclic structure is preferred. Where one or more R groups in such a structure includes an aromatic group, each may be further substituted in one or more positions as described above. Unsubstituted or substituted benzyl substituents are particularly preferred in such structures.
  • the R 2 substituent, or alternatively the R 1 substituent may comprise, in addition to a portion containing a group of the form R as described above, an amino acid structure chosen from various amino acid residues as well as analogs thereof.
  • the structure -Val-R 3 set forth above defines a carboxyl-substituted form of the amino acid valine, wherein the carboxyl-substituent is R 3 as defined above. Where R 3 is chosen to be hydroxyl
  • R 3 may be chosen to be an esterifylng or amidatinq group (as in, e.g., -Val-OR, -Val-NH 2 or -ValNHR 2 ), wherein R is chosen as described above and is preferably a lower alkyl or single-ring aryl (e.g., benzyl or phenyl) group.
  • R 2 (or R 1 ) groups of the form -Ala-R 3 , -Leu-R 3 , -lleR 3 , -Nle-R 3 and -Thr-R 3 refer to the carboxyl- substituted amino acids alanine, leucine, isoleucine, norleucine, and threonine, wherein R 3 may be hydroxyl or other substituent as described above.
  • R 2 and as used above with respect to R 2 (or R 1 ) define carboxyl-substituted serine and threonine moieties having, additionally, a substituent R 4 on the oxygen of the amino acid side chain. Serine-derived moieties are particularly preferred in this position.
  • R 4 in the above formula is chosen to be hydrogen, and R 3 is hydroxyl, a simple serine or threonine moiety results.
  • R 2 (or R 1 ) represent analogs of serine wherein the side chain and/or the carboxylate group of the amino acid has been modified or truncated.
  • Serine is especially preferred as the R 2 substituent, particularly when substituted at R 3 and/or R 4 (most preferably in both positions) with a bulky substituent for R therein.
  • the substituent R 4 on the side chain position be selected from bulky substituents including, for example, benzyl, p-chlorobenzyl, benzyloxymethyl, trityl (triphenylmethyl), t-butyl, cinnamyl, other substituted benzyl, substituted methyl, alkyl, cycloalkyl, phenyldimethylsilyl, t-butyldimethylsilyl and triisopropylsilyl groups, with the benzyl group (forming a side chain ether functionality) being especially preferred.
  • Acyl substituents, including lower acyl substituents such as acetyl groups are also useful.
  • the linking moiety Y is preferably selected in conjunction with the other portions of the claimed compounds (most particularly the optional group Z, and the methylene moieties enumerated by the integer n) in a manner which provides suitable spacing between the guanidinyl (or guanidinyl-analog) group and the carbonyl-containing group designated as -COR 1 .
  • the linking moiety Y is of a hydrocarbon chain form
  • the preferred length of the linking portion of the chain is from about 2 to about 5 methylene or other atomic residues (noninclusive of carbons or other groups appended as branches or substituents to the linking portion of the alkyl chain).
  • Simple alkylene chains of the form -(CH 2 )- r , wherein r is an integer of from 1 to about 8, and preferably 1 to about 5 where the optional Z group is not present, are preferred for the linking group Y.
  • Such chains may be used to yleld des- ⁇ -amino forms of arginine and homologs thereof in the structures of the present compounds.
  • the "linking portion" of Y refers to that portion of Y which effectuates the intramolecular spacing between the two residues of the compound adjacent to the Y group, i.e., the guanidinyl (or guanidinyl-analog) residue bonded to the left of Y and the
  • portions of Y which do not contribute to this inter-residue spacing within the subject compounds such as branch groups or substituents as in, for example,
  • the linking portions affording spacing between the adjacent residues in the compound comprise, respectively, 4 linking atoms (-C-O-C-C-) and 5 linking atoms (-C-C-C-C-C-).
  • the number of such linking atoms, and the resultant spacing distance provided by the linking moiety Y may be modified in conjunction with other portions of the compound so as to provide desired intramolecular spacing as discussed hereinafter.
  • Unsaturated linking moieties Y containing double and/or triple bonds will also preferably include from about 2 to about 5 carbon atoms in the linking portion of the chain, and may include from about 1 to about 3 double or triple bonds.
  • one or more heteroatoms such as O, S or N may be included as substituents within the structure of the linking moiety, as for example to form an ether, thioether or secondary or tertiary amino linking structure.
  • the compounds of the invention do not include those wherein Y is substituted with a primary amino group so as to form (in conjunction with the adjacent portions of the compound) an arginine (Arg) residue.
  • the linking moiety Y includes a cycloalkyl or aryl portion within its structure, such a moiety may be of the form
  • Y 1 is carbon or an O, S or N heteroatom
  • Y 2 is an optional substituent of the form discussed above with respedto substituents on R, espedally hydroxyl, amino or lower (C 1 to C 8 ) aikoxyl or alkyl, or other pharmaceutically suitable substituents such as nitro or halogen (especially chloro or bromo) groups.
  • the integers p and q range independently from 0 to about 5, preferably 0 to 3, and most preferably the sum of p and q is 2.
  • cyclic structures for Y may be made in conjundion with other strudures in the compound so as to achieve a desired intramolecular spacing between the guanidinyl residue and the remainder of the molecule.
  • the cyclic portion of each Y group depided above contributes spacing equivalent to about 3-4 straight-chain methylene units; other cyclic or alternative linking strudures may likewise be evaluated to ascertain their contribution to intramolecular spacing.
  • the optional group Z where present, provides a linking function in addition to that of group Y and, therefore, may also be chosen in conjunction with the other structures in the compound to afford a desired intramolecular spacing. It is most preferred that Z be absent, particularly where the linking moiety Y contains about
  • a preferred Z group is the glycine residue
  • linking portion comprising 3 linking atoms (-N-C-C-).
  • Ala ⁇ ine (Ala), sarcosine (Sar) and ⁇ -alanine ( ⁇ -Ala) are also preferred in this position, and these provide 3 (Ala and Sar) or 4 ( ⁇ -Ala) linking atoms in their strudures.
  • Each of these examples provides, as is preferred, an amino group at the left-hand terminus of Z and a carbonyl group at the right-hand terminus, thereby allowing peptide-like amide bonding to each adjacent portion of the compound.
  • Z is absent in the compound, it is desirable to increase the lengths of the linking portions of the Y group and/or the methylene moieties enumerated by n so as to maintain the desired intramolecular spacing.
  • R 1 in the above strudure be of the form -OR, where
  • R is selected from hydrogen or, more preferably, from bulky substitutents including benzyl, phenyl, and others described above with resped to R 1 , R 2 , R 3 and R 4 .
  • guanidinyl (or guanidinyl-analog) functional group at the left-most position in the molecule is separated from the R 1 -attached carbonyl group by a linking structure that may be depided as
  • the total number of linking atoms in the linking portion of thiS structure is equal to L Y + L Z + n + 3, where L Y and L Z are the number of linking atoms (or their equivalent) in Y and Z, respectively, and n is an integer as defined above. It is generally preferred that L tot be in the range of from about 7 to about 20, and most preferably from about 9 to about 12 Those practicing the invention will, in view of the present disdosure, be able to choose suitable linking groups so as to attain a desired intramolecular spacing for the produd compounds.
  • Y and/or Z represent simple alkylene linking groups
  • R 1 , R 2 and (if present) R 3 and R 4 are esterifylng substituents.
  • the latter compound is especially preferred.
  • compounds of the form are also preferred.
  • a second group of compounds of the invention has the structure
  • X is selected from NR, S and O;
  • Y is an unsubstituted or substituted, linear or branched linking moiety selected from saturated and unsaturated hydrocarbon groups containing from 1 to about 15 linking atoms, and optionally containing one or more heteroatoms and/or one or more cydic structures;
  • R 1 is a substituent selected from -OR (including hydroxyl), -NR 2
  • R 2 is is selected from -R, acyl groups -COR, alkyloxycarbonyl groups of the form
  • R 4 and R 6 are substituents independently selected from groups of the form -R and acyl groups -COR, and R 6 may additionally be selected from alkoxycarbonyl groups of the form
  • linking moiety is a suitable linking moiety, preferably selected from those of the form wherein
  • Y 1 comprises carbon or an O, S or N heteroatom
  • Y 2 is an optional substituent selected from hydroxyl, amino, lower (C 1 -C 8 ) alkoxyl and alkyl, nitro and halogen moieties;
  • p and q are independently integers of from 0 to about 5;
  • n and (where present) m are independently 1 , 2 or 3; and wherein each R is individually a pharmaceutically suitable substituent group, preferably one selected from hydrogen, from linear and branched, unsubstituted and subtstituted C 1 -C 8 lower alkyls, C 2 -C 8 alkenyls, C 2 -C 8 alkynyls, C 6 -C 14 aryls, C 7 -C 14 alkaryls, C 7 -C 14 aralkyls and C 3 -C 14 cycloalkyls, from heteroatomic groups and, in the case of -NR 2 , from cyclized groups forming (in attachment with the nitrogen atom) a 5-8 membered heterocyclic ring optionally containing oxygen, nitrogen or sulfur as a further ring heteroatom.
  • each R is individually a pharmaceutically suitable substituent group, preferably one selected from hydrogen, from linear and branched, unsubstituted and subtstituted C 1
  • the compounds of the invention further include pharmaceutically acceptable base or acid-addition salts of the compounds of Formula II.
  • the pharmaceutical compositions of the invention include such compounds and salts thereof formulated with a pharmaceutically acceptable excipient. It will be seen that the compounds of Formula II are strudurally similar to those of
  • D-enantiomeric residues be utilized in the reversed-orientation residues, i.e., in the Asp residue (to the right of Z') and, if present, in any optically active site in R 5 .
  • the other preferred substituents and preferred structures described above for Formula I are likewise preferred for the compounds of Formula II.
  • R 5 substituent is that of the substituted or unsubstituted reverse form of serine, i.e.,
  • the linking group Z' is preferably a lower diaminoaikane structure, e.g., of the form -NH(CH 2 ) m -NH-, where m is (as defined) 1 , 2 or 3.
  • Y is preferably a 1 to 8 carbon methylene chain as discussed above for Formula I.
  • the Y and Z' moieties in Formula II are conveniently selected so as to maintain a desired intramolecular spacing in the compound between the guanidinyl (or guanidinyl-analog) group and the carbonyl-containing group designated as -COR 1 .
  • the total number of linking atoms in the linking portion of Formula II (L tot ) is equal to L Y + L Z + n + 2, where L Y and L Z are the number of linking atoms (or their equivalent) in Y and Z', respectively, and n is the integer as defined above.
  • L tot be in the range of from about 7 to about 20, and most preferably from about 9 to about 12.
  • the title compound was prepared by first dissolving 1.02 g (1.72 mmole) of the starting ester
  • NMM N-methylmorpholine
  • condensations may be performed in DMF with DCC as the condensing agent.
  • the amino acid DCHA salts are neutralized by suspending them in 1M aqueous NaHSO 4 and repeatedly extracting with ethyl acetate. The combined extrads are dried over Na 2 SO 4 , filtered, and evaporated.
  • the BOC protecting groups are removed when needed by dissolving the amine in ethyl acetate and adding three equivalents of HCl in the same solvent. When the cleavage is complete, the volatiles are removed at reduced pressure and the residue dried by adding toluene and removing it aqain on the rotary evaporator.
  • the hydrochloride salt is neutralized in situ by adding NMM to the DMF reaction mixture. After the condensation is complete the dicydohexylurea is removed by filtration and the volatiles removed at reduced pressure. The peptide residue is taken up in ethyl acetate, washed with aqueous citric acid, washed with aqueous sodium bicarbonate, dried over anhydrous sodium sulfate, and evaporated at reduced pressure.
  • the depided compound ⁇ -guanidinylpropionyl-glycyl-glutamyl(gamma-methyl ester)-threonine ( ⁇ -triisopropyl silyl ether) (3), is prepared according to the following steps.
  • BOC-glutamyl(gamma-methyl ester)-threonine methyl ester The BOC-protected dipeptide is prepared according to standard procedures by sequential solution-phase coupling of BOC-Glu(gamma-methyl ester) and Thr methyl ester.
  • BOC-glycyl-glutamyl(gamma-methyl ester)-threonine methyl ester This BOC-protected tripeptide is prepared from the foregoing deprotected compound by coupling BOC-Gly with DCC in DMF/NMM in the usual fashion.
  • the protected tetrapeptide above is dissolved in dichloromethane and one equivalent of piperidine is added. When deavage of the FMOC group is complete, the volatiles are removed at reduced pressure. The resulting free amine is immediately dissolved in acetonitrile and one equivalent of aminoiminomethane sulfonic acid is added. The desired guanidine compound is colleded and dried.
  • the reaction mixture is diluted with ethyl acetate, washed with aqueous citric acid, washed with aqueous sodium bicarbonate, dried, and filtered to give a clear solution.
  • the organic solution is shaken with 50 psi of hydrogen and palladium on carbon until the ⁇ -benzyl ester is gone.
  • the solution is filtered through a Celite pad and one equivalent of dicyclohexylamine is added.
  • 6-Ami ⁇ o-3(R)-5-dihydroxyl hexnoic acid ethy ester 3-t- butydimethylsily ether The epoxides from above are treated with t-butyldimethylsilyl chloride and imidazole in DMF under the usual conditions.
  • the crude silyl ether is dissolved in ethanol and treated with sodium azide and amonium chloride. After the epoxide has been consumed, the reaction mixture is shaken under 50 psi of hydrogen in the presence of platinum oxide.
  • the crude amine solution is filtered, diluted with ethyl acetate, and washed with aqueous bicarbonate solution.
  • 1-BOC derivative of piperazine is prepared as described by Ca ⁇ ino in J. Org. Chem., 1983, 48, 611 and condensed with the ester from the preceding step using the DCC general procedure.
  • the resulting amide is purified by flash chromatography on silica gel.
  • 5-Guanidinylpenta ⁇ oic acid A solution of 5-aminopentanoic acid and one equivalent of potassium carbonate in water is stirred as one equivalent of aminoimino methane sutfonic acid is added in portions over 15 minutes. The resulting slurry is stirred at room temperature for 24 hours.
  • the predpitate is colleded and dried to give the desired product 1-(5-Guanidinylpentanoyl)-4-(N,N-diethyl-D,L-valyl-D- ⁇ -benzylaspartyl)-piperazine
  • N'-BOC-ethylenediamine This compound is prepared according to the procedure of Callahan, J. Med. Chem., 1989, 32, 341.
  • N-BOC-D-Glutamic Acid Y-3,5-dinitrobenzyl ester
  • the produd is purified by column chromatography to give the title compound.
  • the 4-ureidobutyric acid and one equivalent of N'-BOC-ethylenediamine from above are condensed with the general DCC coupling procedure described above.
  • the produd is deprotected with 4 N HCl in ethyl acetate for 2 hours and neutralized with DIEA.
  • the produd is deprotected as usual and coupled with N-BOC-D-Glu (gamma-3, 5-dinitrobenzyl ester), above, using the general DCC method.
  • the produd is deprotected and coupled to BOC-D-homoSer(Bzl), above, in an analogous way.
  • This produd is deprotected, neutralized, and coupled to the symmetric anhydride of phenylacetic acid (Aldrich).
  • the final purification by reversed phase HPLC and tyophiiization affords the desired compound, (6). * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
  • Purification of the compounds of the invention may be achieved using methods known in the art, including reversed phase HPLC size exclusion chromatography, partition chromatoqraphy on polysaccharide gel media such as Sephadex G10 or
  • G25 or counter current distribution.
  • fluoride which if present in even small amounts may alter the biological profile of the compound.
  • ion exchange chromatoqraphy using AG 3 -4X acetate for example, to exchange the fluoride salt, may be used.
  • the subject compound is dissolved in water and passed over an acetate-form resin.
  • the eluate is colleded, and lyophilized to dryness.
  • Salts of carboxyl groups of the produd compounds may be prepared in the usual manner by contacting the compound with one or more equivalents of a desired base such as, for example, a metallic hydroxide base such as, for example, sodium hydroxide; a metal carbonate or bicarbonate base such as, for example, sodium carbonate or sodium bicarbonate; or an amine base such as, for example, triethylamine, triethanolamine, and the like.
  • a desired base such as, for example, a metallic hydroxide base such as, for example, sodium hydroxide
  • a metal carbonate or bicarbonate base such as, for example, sodium carbonate or sodium bicarbonate
  • an amine base such as, for example, triethylamine, triethanolamine, and the like.
  • the pH should be kept below approximately 8 in order to avoid complications such as racemization, deamidation, degradation or other undesirable side readions.
  • Acid salts of the compounds may be prepared by contacting the compound with one or more equivalents of the desired inorganic or organic acid, such as, for example, hydrochloric acid, acetic acid or citric acid.
  • Esters or amides of carboxyl groups of the compounds may be prepared by any of the usual means known in the art for converting a carboxylic acid or precursor to an ester or amide. * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
  • an effective amount of the active compound, including derivatives or salts thereof, or a pharmaceutical composition containing the same, as described below, is adminiatered via any of the usual and acceptable methods known in the art, either singly or in combination with another compound or compounds of the present invention or other pharmaceutical agents such as immunosuppressants, antihistamines, corticosteroids, and the like.
  • compositions can thus be administered orally, sublingually, topically (e.g., on the skin or in the eyes), parenterally (e.g., intramuscularly, intravenously, subcutaneously or intradermally), or by inhalation, and in the form of either solid, liquid or gaseous dosage including tablets, suspensions, and aerosols, as is discussed in more detail below.
  • the administration can be conducted in single unit dosage form with continuous therapy or in single dose therapy ad libitum.
  • Useful pharmaceutical carriers for the preparation of the pharmaceutical compositions hereof can be solids, liquids or gases; thus, the compositions can take the form of tablets, pills, capsules, powders, enterically coated or other protected formulations (such as by binding on ion exchange resins or other carriers, or packaging in lipid protein vesicles or adding additional terminal amino acids), sustained release formulations, solutions (e.g., ophthalmic drops), suspensions, elixirs, aerosols, and the like.
  • Water, saline, aqueous dextrose, and glycols are preferred liquid carriers, particularly (when isotonic) for injectable solutions.
  • the carrier can be selected from various oils including those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like.
  • suitable pharmaceutical excipients include starch, cellulose, talc, glucose, ladose, sucrose, gelatin, matt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.
  • compositions may be subjected to conventional pharmaceutical expedients such as sterilization and may contain conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, buffers, and the like.
  • conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, buffers, and the like.
  • Suitable pharmaceutical carriers and their formulations are described in Martin, Remington's Pharmaceutical Sciences, 15th Ed. (Mack Publishing Co., Easton 1975) (see, e.g., pp. 1405-1412, 1461-1487).
  • Such compositions will, in general, contain an effective amount of the active compound together with a suitable amount of carrier so as to prepare the proper dosage form for proper administration to the host
  • the therapeutic methods of the present invention are practiced when the relief of symptoms is specifically required or perhaps imminent; in another preferred embodiment, the method hereof is effectively practiced as continuous or prophyladic treatment.
  • the particular dosage of pharmaceutical composition to be administered to the subject will depend on a variety of considerations including the nature of the disease, the severity thereof, the schedule of administration, the age and physical characteristics of the subject, and so forth. Proper dosages may be established using clinical approaches familiar to the medicinal arts. It is presently believed that dosages in the range of 0.1 to 100 mg of compound per kilogram of subject body weight will be useful, and a range of 1 to 100 mg per kg generally preferred, where administration is by injection or ingestion. Topical dosages may utilize formulations containing generally as low as 0.1 mg of compound per ml of liquid carrier or excipient, with multiple daily applications being appropriate.
  • the compounds and therapeutic or pharmaceutical compositions of the invention are useful in the study or treatment of diseases or other conditions which are mediated by the binding of integrin receptors to ligands, including conditions involving inappropriate (i.e., excessive or insufficient) binding of cells to natural or other ligands.
  • Such diseases and conditions include inflammatory diseases such as rheumatoid arthritis, asthma, allergy conditions, adult respiratory distress syndrome, inflammatory bowel diseases (e.g., ulcerative colitis and regional enteritis) and opthalmic inflammatory diseases; autoimmune diseases; thrombosis or inappropriate platelet aggregation conditions; reocclusion following thrombolysis; cardiovascular disease; neoplastic disease including metastasis conditions; as well as conditions wherein increased cell binding is desired, as in wound healing or prosthetic implantation situations as discussed in more detail above.
  • inflammatory diseases such as rheumatoid arthritis, asthma, allergy conditions, adult respiratory distress syndrome, inflammatory bowel diseases (e.g., ulcerative colitis and regional enteritis) and opthalmic inflammatory diseases; autoimmune diseases; thrombosis or inappropriate platelet aggregation conditions; reocclusion following thrombolysis; cardiovascular disease; neoplastic disease including metastasis conditions; as well as conditions wherein increased cell binding is desired, as in wound healing or pros
  • derivatives of the present compounds may be useful in the generation of antigens which, in turn, may be useful to generate antibodies.
  • These antibodies will, in some cases, themselves be effective in inhibiting cell adhesion or modulating immune activity by acting as receptors for matrix proteins or other cell adhesion ligands, or, if antiidiotypic, by acting to block cellular receptors.
  • the following assay established the activity of the present compounds in inhibiting cell adhesion in a representative in vitro system.
  • the assay was a competition assay in which both fibronectin and a test compound were present Microtiter plates were first precoated with fibronectin. The test compound was then added in increasing concentrations with cells known to contain the fibronectin receptor. The plates were then washed and stained for quantitation of attached cells.
  • the present assay directly demonstrates the anti-cell adhesion activity and modulatory activity of the present compounds. For example, by immobilizing the compound on a surface, one could adhere appropriate cells to that surface. Other cell adhesion modulation activity, and utilities pertinent thereto, will be apparent to those skilled in the art.
  • the cell line U937 was purchased from American Type Tissue Culture Collection. The cells were cultured in RPMI media (J.R. Scientific Company, Woodland Hills,
  • Fibronedin was purified from human plasma according to the procedure of Engvall, E. and Ruoslahti, E., Int J. Cancer 1977, 20, 1-4.
  • Microtiter plates (96-well, Falcon) were coated overnight at 4oC with 5 ⁇ g/ml fibronedin (FN) (for a total volume of 0.1 ml) or, as a control, 5 ⁇ g/ml bovine serum albumin (BSA) diluted in phosphate buffered saline (PBS, 0.01 M NaPO 4 in 0.9% NaCl at pH 7.2 to 7.4). Unbound proteins were removed from plates by washing with PBS. The plates were then coated With 100 ⁇ l of PBS containing 2.5 mg/ml BSA for one hour at 37oC. This procedure is a modification of a previously published method, Cardarelli, P.M. and M.D.
  • FN fibronedin
  • BSA bovine serum albumin
  • a U937 culture was collected and washed two times with Hanks Balanced Salt Solution. The cells were counted and adjusted to 1.5 ⁇ 10 6 cells per ml in Dulbecco's Modified Eagles Medium (DMEM) plus BSA (2.5 mg/ml) for cell attachment assay. Subject compounds were then dissolved in DMEM and BSA, and the pH was adjusted to 7.4 with 7.5% sodium bicarbonate. The compounds (100 ⁇ l) were added to FN-coated wells, at 1.5, 0.75, 0.375, 0.188, 0.094, 0.047, 0.023, 0.012, 0.006 and 0.003 mg/ml final concentration and U937 cells (100 ⁇ 1) were added per well.
  • DMEM Dulbecco's Modified Eagles Medium
  • BSA 2.5 mg/ml
  • Subject compounds were then dissolved in DMEM and BSA, and the pH was adjusted to 7.4 with 7.5% sodium bicarbonate. The compounds (100 ⁇ l) were added to FN-coated well
  • Results The table below shows the results of the cell adhesion inhibition assay for representative compounds of the invention. Potency is expressed in ⁇ M units.
  • the accompanylng figure is a diagram representing the curve of cell adhesion inhibition for the compound

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Abstract

Sont décrits des composés contenant du guanidinyle, ainsi que des composés et des sels apparentés, utiles comme antagonistes de récepteurs cellulaires pour moduler l'adhésion cellulaire par l'intermédiaire de récepteurs d'intégrine et/ou de récepteurs de fibronectine. Sont également décrits des procédés pour synthétiser, tester, formuler, et utiliser les composés comme agents thérapeutiques.
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