JP2003509468A - Methods and compositions for inhibiting adhesion formation - Google Patents

Abstract

  (57) [Summary] Compositions and methods for inhibiting or ameliorating post-operative / post-wound adhesion formation in a patient are disclosed. Such compositions and methods utilize antagonist molecules that prevent or inhibit the binding of alphaVbeta3 integrin to extracellular matrix proteins such as fibronectin.

Description

Detailed Description of the Invention

STATEMENT OF GOVERNMENT RIGHTS The work described herein was supported in part by NIH grant HD34824. The US Government may have certain rights in this invention.

[0002]     Field of the invention   The present invention relates to the field of medicine and inhibition of post-surgical / post-wound adhesion formation.

BACKGROUND OF THE INVENTION Surgical intervention involves wounding a patient to provide treatment. One undesirable outcome resulting from surgery is postoperative adhesion formation. The term "adhesion" used in the medical sense refers to a gluing, the process of two surfaces or sites adhering or joining. For example, the joining of opposing surfaces of the wound or the peritoneum. Also, a plurality of adhesions can refer to an inflammatory zone connecting opposing serous surfaces. The term adhesions, as used herein, also includes fibrinous adhesions, which are adhesions composed of fine threads of fibrin that result from the exudation of plasma or lymph, or extravasation of blood. Keloids, a smooth hyperplasia of fibroblast tissue that occasionally or naturally occurs at the site of injury, is also a type of adhesion.

The development of adhesions has been reported to be a major source of post-operative morbidity and lethality. The most frequently performed surgical procedures involving significant adhesion formation are gynecological, cardiovascular and abdominal general surgery. This applies not only to laparoscopic surgery, but also to conventional surgery.

Complications of intraabdominal adhesions include intestinal obstruction, chronic or recurrent pelvic pain, female infertility, and extended surgical time (when additional surgery is needed) and postoperative complications. In extreme cases, debilitating adhesions can be treated by adhesion dissection, surgical incision or lysis of adhesions (adhesion lysis). Therefore, it would be extremely useful to have methods and compositions that inhibit adhesion formation in patients.

Animal surgery has become more popular as the value of animals both emotionally and economically has increased with veterinary skills and practices. Animals suffer the same problems with regard to postoperative adhesion formation and its debilitating effects. Sufficient and humane consideration for surgically treated animals has facilitated the adoption of many techniques and procedures from human diagnostics and surgery for use on animals. Therefore, it would be useful to have methods and compositions that inhibit adhesion formation in animals.

When the majority of adhesions begin to form due to the expression of a fibrin matrix between tissue surfaces, physical barrier agents have been tested to prevent adhesion formation by limiting tissue attachment during peritoneal healing. It was Generally, physical barrier agents, either membranous or gel-like, have been used to reduce attachment of injured peritoneum until peritoneal regeneration occurs and thus inhibits or prevents adhesion formation. In the rat model system, peritoneal regeneration takes approximately 7 days. A rat model system was found to require a barrier agent placement of at least 36 hours to prevent adhesion formation (Harr.
Is, et al., 1995, Surgery, 117: 663-9).

Although numerous adjuncts for the prevention of adhesions have been tested in animal models, currently only three barrier agents are approved for use in humans to reduce postoperative adhesion formation. . Interceed ™ (Johnson & Johnson Me)
(dical) is an oxidized regenerated cellulose, and Seprafilm (trademark) (G
enzyme Corp. ) Is modified hyaluronic acid complexed with modified carboxymethyl cellulose and Preclude ™ (W.L. Gore) is expanded polytetrafluoroethylene. Interceed ™ is suitable for use in the absence of blood. Preclude ™ is approved as a pericardial substitute, but it is not bioabsorbable. Seprafilm ™
Is licensed for use in abdominal wall and hysterotomy.

Potential anti-adhesion agents tested in the rat system include lactated Ringer's solution, 32% dextran 70 solution, and 1% and 2% modified carboxymethylcellulose solution. Half of each solution was applied to the defect and the excess was pooled in the peritoneal cavity of the abdomen. Lactated Ringer's solution was not effective because it was rapidly absorbed in the peritoneal cavity and therefore was not present for the critical 36 hours. Each of the other slightly more viscous solutions was more effective.

Hyaluronic acid (HA) has been used as a barrier agent in abdominal pelvis and orthopedics. Modified HA as an absorbent gel, and hyaluronan-based gels of self-cross-linking polysaccharide (ACP) have also been tested. ACP hyaluronic acid derivatives have been shown to prevent laparoscopic adhesion formation in a rabbit model system, DeIaco et al., 1998,
Fertility and Surgery, 69 (2): 318-323.

Chemically modified hyaluronate and carboxymethyl cellulose (HA / CMC)
Gel formulations are described by Leach et al., 1998, Fertility and Su.
rgery, 69 (3): 415-418 in a rabbit model system.

Evaluation of polyethylene glycol / polylactic acid (EG / LA) films for preventing adhesion formation in a rabbit model system was performed by Rodgers et al., 1998, Fert.
Ility and Surgery, 69 (3): 403-408.

Accordingly, there is still a need in the medical community for materials and methods useful in inhibiting or preventing adhesion formation not only in humans but also in animals.

SUMMARY OF THE INVENTION The present invention provides needed compositions and methods useful for inhibiting or ameliorating adhesion formation in mammals, including humans.

Adhesions resulting from a wound or surgical procedure are treated in the mammal by treating the wound or surgical site with an antagonist molecule that interacts with the alpha V beta 3 (αvβ3) integrin or integrin binding site of the extracellular matrix protein. It can be inhibited or at least ameliorated by blocking the binding of αvβ3 integrin to extracellular matrix proteins in the body. Antagonist molecules suitable for this purpose are extracellular matrix proteins such as αvβ3 on fibronectin.
Proteins, peptides (linear and cyclic) and peptidomimetics that mimic the integrin binding site, or bind to the αvβ3 integrin itself, thereby interfering with its binding to extracellular matrix proteins. Such αvβ
Examples of 3 antagonist molecules include αvβ3 binding sites on extracellular matrix proteins that mimic or block, or by specifically binding to an antigenic epitope of the αvβ3 molecule, It is a monoclonal antibody and its biologically active site or fragment that realizes inhibition of binding to extracellular matrix proteins. One such monoclonal antibody is the complete form of LM609, as well as biologically active antigen binding sites or fragments such as Fab, Fab2, Fv, and mixtures thereof. Mouse and humanized forms of monoclonal antibody LM609 are also suitable for the purposes of the invention.

Furthermore, it contains an amino acid residue sequence corresponding to the αvβ3 integrin binding site on fibronectin, for example the amino acid residue sequence Arg-Gly-Asp-Ser.
(RGDS) or its bioequivalent fibronectin fragment as an αvβ3 antagonist molecule suitable for the purposes of the present invention, wherein αvβ3 integrin is a fibronectin or amino acid residue sequence RGDS or its biological equivalent. Can be used to prevent binding to any other extracellular matrix protein having a binding site defined by

So-called peptidomimetics, ie non-peptidic organic compounds, which mimic the binding of αvβ3 integrin with extracellular matrix proteins, such as fibronectin, by mimicking the peptides with respect to their interaction with αvβ3 integrin are also suitable. .

Adhesion formation refers to an adhesion-inhibiting dose of at least one said αvβ3 antagonist molecule,
Application, either directly or in a physiologically compatible carrier, to the surgical site is inhibited or at least minimized. Application of these antagonists is easily accomplished via intraperitoneal (ip), subcutaneous (sc) injection, intravenous (iv) administration, or other suitable route of administration. To be done.

Suitable carriers may be liquid as well as physiologically acceptable absorbable pastes and solids. Usually at least about 50 micrograms per milliliter (μ
The antagonist molecule can be applied as an antagonist composition with the antagonist molecule dissolved in the aqueous carrier at a concentration of (g / ml) or present as a suspended substance therein. Similarly, the antagonist composition may be an absorbable paste or solid composed of its inhibitor molecules in solution or suspension, and an absorbable gelatin sponge or powder, or an absorbable dry hydrogel as a carrier, an absorbable absorbable hydrogel. It may be a hyaluronic acid derivative or the like. The αvβ3 antagonist composition can be packaged in an appropriately sized dosage form with a label indicating that the inhibitor composition contained in the container can be used to inhibit adhesion formation.

The αvβ3 antagonists contemplated by the present invention include extracellular matrix proteins or αvβ3
It can bind to any of the integrin molecules. However, in order for such binding to be effective, the normal interaction between the αvβ3 integrin and its binding site on the extracellular matrix protein must be inhibited. Examples of such extracellular matrix proteins include fibronectin, fibrinogen, vitronectin, von Willebrand factor, laminin, collagen, tenascin, osteopontin, thrombospondin and the like.

A preferred method of inhibiting post-surgical adhesion formation involves administering to a surgical patient (human as well as veterinary) an adhesion-inhibiting amount of an αvβ3 integrin inhibitor molecule. Such administration may be carried out directly or as an aerosol spray, or as a suitable carrier, via a pad, gel, solution, suspension, etc., in which a dose of the inhibitor composition is applied to the tissue surface to be protected from adhesion formation. Including administration.

Detailed Description of the Preferred Embodiments Adhesion formation occurs as an abnormality of the wound healing cascade involving fibroblast cell adhesion and migration. Adhesions often occur as a result of trauma or bleeding during surgery when obstruction of the peritoneal surface exposes the underlying stromal layer. This exposure then releases quinines and histamine, followed by increased capillary permeability and the release of serous blood exudate containing inflammatory cells. This fibrin-rich exudate causes coagulation on the surface of the lesion. With the clot undissolved, inflammatory cells and fibroblasts infiltrate the fibrin-rich extracellular matrix, resulting in the formation of adhesions.

The extracellular matrix is composed of an interactive network structure of proteins forming a mesh structure to which cells adhere for tissue formation. The macromolecules that make up the extracellular matrix are secreted primarily locally by the cells within the matrix. In most connective tissues, these macromolecules are mainly secreted by fibroblasts or fibroblast-based cells such as chondroblasts, osteoblasts. The two main groups of extracellular macromolecules that make up the matrix are the glycosaminoglycans (GAGs), a polysaccharide known to be covalently linked to proteins, usually in the form of proteoglycans, and fibrous proteins. . Fibrous proteins are generally considered to be either of two functional types, predominantly structural (eg, collagen and elastin) or predominantly adhesive (eg, fibronectin and laminin). For example, The Molecular Biology of the Cell, 2nd ed. (Alberts
Et al., Editors, Garland Publishing, Inc. , Ne
w York, 1989) 802-804.

[0024]   Cell adhesion is a function of cell migration, proliferation and growth in embryonic tissue and extracellular matrix.
And malignant tumor formation, inflammation, immune regulation and hemostasis
Contribute. Mediators of cell adhesion, transmembrane cell receptors, integrins, immunoglobins
Brin superfamily, cadherin, selectin, CD-44 related molecules and
And transmembrane proteoglycans. (Chothia & Jones, 1997, Ann. Rev. Biochem. , 66: 823-62). I
Integrins are a group of important binding proteins that interact with multiple ligands.
αvβ3 integrin is a membrane-bound membrane recognized as important for cell-cell adhesion and migration.
It is a glycoprotein. Integrins are extracellular matrix components, cell surface immunoglobins.
Brin (Ig) superfamily receptors, microbial surface components, and certain blood
It binds to a variety of ligands including serum proteins. For a review, see Loftus, J. et al. C
． Etc., 1994,J. Biol. Chem., 269 (41): 25235-2.
See 5238.

The mouse IgG monoclonal (mAb) antibody LM609 produced by the hybridoma cell line LM609 is specific for αvβ3 integrin (Ch
eresh et al., 1987, J. Am . Biol. Chem. , 262: 17703-1
7711). The mouse hybridoma LM609 was introduced on September 15, 1987 by the American Type Culture, an international depository under the Budapest Treaty.
e Collection (ATCC, Rockville, MD, USA) and has been assigned the ATCC name HB9537.

LM609 complementary DNA was cloned and its soluble Fab portion was produced from transformed host cells. In order to reduce its immunogenicity, the LM609 antibody has been humanized (WO 99/29888).

Proteins and peptides suitable for use as αvβ3 antagonist molecules are those that include the αvβ3 complementary binding site on fibronectin, the amino acid residue sequence RGDS, or their bioequivalents. The peptide may be linear or cyclic.

Also suitable are non-toxic, non-peptidic organic compounds that define a region substantially complementary to the αvβ3 integrin binding site or the binding site for αvβ3 integrin on extracellular matrix proteins.

When used to inhibit adhesion formation, the αvβ3 integrin antagonist molecule may be absorbed powder, alone or in paste form, either directly or as an aerosol powder, or in a physiologically compatible carrier which may be a liquid such as water. Together, it can be applied to the surgical site. As such an absorbent powder, GELFOAM (registered trademark) (Upj
ohn Co. Examples thereof include sterile gelatin powders marketed under the name of (1). Alternatively, the αvβ3 integrin antagonist molecule is delivered to the surgical site as a sterile gelatin foam or sponge, a dry absorbable hydrogel of the type described in US Pat. No. 5,409,703 to McAuley et al., Or a hyaluronic acid derivative. be able to. For intraperitoneal (ip), subcutaneous (sc) or intravenous (iv) administration, the preferred carriers are water, aqueous media such as saline.

The rabbit sidewall model of adhesion formation has been previously described (Rogers et al., 1).
996, J.I. Invest. Surg. , 9: 388-91; Rogers et al.,
1998, above). 55 mg of ketamine hydrochloric acid and 1 k per 1 kg of rabbit body weight
Rabbits were anesthetized by intramuscular injection with a mixture of 5 mg xylazine per gram. After preparation for aseptic surgery, midline laparoscopic surgery was performed. Acupressure was applied to temporarily expose the cecum and intestines and cause subserosal hemorrhage over all surfaces that are believed to be in contact with the sidewall lesion area. The injured intestine was then lightly abraded with 4 inches of 4 × 4 heavy sterile gauze until punctate bleeding was noted. The cecum and intestines were then returned to their normal anatomical position.

This rabbit model closely resembles the standard rat model of adhesion formation described by Harris et al. (1995, supra). In this rat model, abdominal wall defects and cecal abrasions were created, air dried for 10 minutes, and placed in contact with the two injured surfaces before closing. Intraperitoneal sodium pentobarbital (43 mg / k
Rats were anesthetized under g). Prepare the abdominal abdomen and sterilize with iodine, 70%
Rinse with alcohol. A 6 cm incision was made on the skin midline, and the skin contracted. 4 midline of abdominal wall
A cm incision was made and the right abdominal wall was rejected. A 1 × 2 cm section of the parietal peritoneum was laterally separated 1 cm from the midline incision and was sharply excised from the wall, including the underlying superficial muscle layer. The cecum was then lifted and positioned so that the cecum would contact the abdominal wall defect when closed. The cecum was then abraded in a standard manner by rubbing with a scalpel blade so that a homogenous surface of petechiae was on the 1 × 2 cm area. The abdominal wall defect was also scratched. Both the abdominal wall and the caecal defect were exposed to air for 10 minutes. Then place both defects in contact,
The midline incision was closed with continuous sutures with 4-0 polypropylene and the skin closed with 4-0 silk (Harris et al., 1995, supra).

Another rabbit system that studies genital adhesions is the rabbit uterine horn model. In this model, a midline incision is made and the uterine horn is brought out through the incision. An area of about 5 cm in length around the entire circumference of the uterine horn is rubbed with a surgical gauze by rubbing it 12 times with a scalpel blade. This injury results in systemic erythema without active bleeding areas. The uterine horn is then replaced in the abdominal cavity and the wound is closed.

With the advent of surgical laparoscopic instruments and techniques, such minimally invasive surgery has become more common. However, adhesion formation is still a possible complication of such laparoscopic surgery. A rabbit animal model has been used to investigate the prevention of laparoscopic adhesions (DeIaco et al., 1998, supra). Briefly, a Velez needle was inserted into the abdominal cavity through the midline abdominal wall using an automatic peritoneal insufflator for the delivery of carbon dioxide gas. The trocar was then inserted through the abdominal wall at the same location. The arthroscope was then inserted intraperitoneally through the trocar using a 300 W xenon light source. All surgical procedures are performed with an endoscopic microcamera. After abdominal necropsy, laparoscopic scissors and atraumatic forceps were inserted through two lateral incisions without a trocar.

A 2 × 2 cm area of the right uterine horn is peeled off with forceps for 30 seconds, and 1 cm is placed in the distal right uterine horn.
A standardized injury to the peritoneum or internal surface can be created by making an incision and scraping a 5 x 5 cm area of the peritoneum of the abdominal wall in front of the previous lesion.

While it is possible to administer the entire mouse LM609 mAb to the surgical site of the patient, a host immune response to the mouse LM609 protein (ie, human anti-mouse antibody HAMA in human patients, feline or canine anti-mouse antibodies and other It will be ruled out for long-term or multiple-use so that no other reaction (in the treated animal) is elicited. This can be partially minimized by using LM609 truncated to either Fab, Fab2, Fv constructs, or mixtures thereof. Production of antibody fragments such as Fab, Fab2, and Fv is known in the art, and for example, French (1998, Methods in Molecular Biology , Immunochemical Protocols,
2nd Ed. , 80: 121-134). It is also known to use recombinant DNA methods to prepare these antigen binding proteins and artificial structures (ie single chain Fv, single chain Fv, single chain Ab; molecular recognition unit MRU). For example, Verhoeyen et al., "Advances in antibo
dy engineering "in Molecular Immunolo gy, (IRL Press at Oxford University P
Ress, Oxford, 1996) chapter 7.

When used in the treatment of humans, the mouse LM609 antibody or portion thereof is used to modify the immunogenic epitopes on the surface of the antibody so that it appears as a human protein to the treated host in order to preserve the amino acid residues in its structural protein. By judiciously substituting groups, it can be humanized, at which time the antigen specificity of the binding active site, including the complementarity determining region (CDR) amino acid residues, is preserved to maintain antigen epitope binding specificity. Has been done. Humanized monoclonal antibodies are known and have been previously described in the art. For example, Waldmann et al. (US Pat. No. 5,502,167) describe a humanized antibody in which the amino acid sequence of the CDRs is derived from the sequence of the CDRs of a monoclonal antibody that has binding specificity for quiescent and activated T cells. ing.
Hogenboom et al. (US Pat. No. 5,565,332) describes a method for producing antibodies with increased human specificity, which involves selective mutation of either heavy or light chains, recombination of mutant chains, and binding activity. Describes a method that requires antigen-selective selection against. Adair et al. (US Pat. No. 5,859,205) describe a specific method for grafting the CDRs of the antibody heavy and light chains onto the receptor framework region. Thus, grafting the amino acid residues of the LM609 antibody encoding active site-forming residues onto a human antibody framework structure, thereby creating a chimeric antibody having the binding specificity of the LM609 antibody but the antigenic appearance of the human antibody. Is possible. A unique method of optimizing CDR grafting that further required chain reassembly and antigen selection was described in WO 99/29888 (Barbas et al.).

Any decrease in affinity that may occur in this process, and thus decrease in binding efficiency,
It can be minimized by sorting with antigen to select better binding structures and producing multivalent binding structures. For example, by using a chemical linker known in the art to crosslink two or more LM609 antigen-binding active sites or humanized active sites, a bifunctional group of the active site that can act more effectively as a blocking agent. Alternatively, it may be advantageous to form polyfunctional groups. Similarly, it is possible to attach multiple active sites to solid supports such as microparticles and larger latex polymers or colloidal beads. The attachment of whole antibodies or active site containing fragments or structures thereof to solid support systems is known in the art. For example, Wang (1998, Methods in Molecule
ar Biology, Immunochemical Protocols,
2nd Ed. 80: 365-376) describes the use of immunomagnetic beads coated with bioreactive molecules such as antibodies, streptavidins, lectins, peptides for cell sorting.

By making antibodies that interfere with or block the binding of αvβ3 integrin to fibronectin or other extracellular matrix proteins, and thus to the formation of the contacts necessary for adhesion development, the method of the invention. Or it is possible to prepare an equivalent composition of the invention. For that purpose, αv
β3 integrin, or α of fibronectin and other extracellular matrix proteins
Appropriate animals can be immunized to stimulate the production of antibodies specific for the vβ3 binding site. Such suitable antibodies can be selected by antigen selection using conventional methods known in the art. It is also well known to generate monoclonal antibodies from properly selected clones, which can be used to produce suitable αvβ3 inhibitor molecules.

Substitution of any of these existing or subsequently generated antibodies in the compositions or methods of the invention is within the scope of the invention and is equivalent in result and means.

In addition to the above, antagonist molecules suitable for inhibiting αvβ3 binding can also be readily selected by routine selection. In Vivo Utilizing Cell Adhesion to Prepared Extracellular Matrix Comprising Fibronectin or Other Suitable Matrix Components
An in vitro screening test can be constructed. In the initial selection system, candidate molecules can be administered and the subsequent behavior of cells scored. Decreased binding to substrates that reflect adhesion formation or inhibition of growth of matrix components represents a promising candidate for further study. To reduce the time and work of sorting by performing batch sorting, the initial sorting can be done with a mixture of compounds. When any initial result indicates success, the specific inhibition of a particular compound mixture can be individually tested.
If desired, such sorting can be fully automated. Regardless of how such inhibitors are discovered, it is envisioned that they will be properly formulated in the compositions of this invention for use in the methods of this invention. The optimal concentration for use with any such compound is ideally at least comparable to the optimal concentration of the LM609 antibody, which minimizes any potential adverse or toxic effects.

Antagonist molecules that specifically bind to or interfere with the αvβ3 integrin binding site of extracellular matrix protein molecules are also suitable for practicing the present invention.

[0042]   The invention and many of its embodiments are illustrated by the following examples.

EXAMPLE An initial study with a solution containing all mouse mAb LM609 at a concentration of 0.1 to 10 mM was performed using the rabbit uterine horn model. The advantage of using a solution for these tests is that, unlike physical interfering agents, there is no need for precise placement of the physical interfering agent or (in some cases) removal of that interfering agent later There is no. The solution can be placed intraperitoneally and thus infiltrate the entire abdominal cavity without restriction or localization. Therefore, administration of a therapeutic composition containing mAb LM609 can be more easily achieved than with the placement of known physical obstacles, and its efficacy is less dependent on physical factors.

Randomly assigned to receive “open and closed” sham treatment (sham group, n = 3) or intentionally create pelvic abrasions with standard methods (experimental group, n = 18) Twenty-one New Zealand white rabbits underwent a laparotomy. The experimental animals were subjected to a midline laparotomy and the sidewalls, bladder, uterus and fallopian tubes were abraded with 200 grit sandpaper until ecchymosis occurred. After the injury, the rabbits in the experimental group were randomly assigned to 2 ml of saline (saline control group; group iii, n = 6), 2 ml of anti-beta1 antibody (anti-beta 1 group; 1 mg / ml, group ii, n). = 6), or 2 ml of LM609 (anti-αvβ3 group; 1 mg / ml, group i, n = 6).
Were administered intraperitoneally. For muscle 3.0 Polyglactin 910 (Vicr
yl) and 4.0 polyglactin 910 to close the skin subcutaneously,
The abdominal wall was closed in two layers.

The place of the animals was changed to a light-dark cycle every 12 hours, 150 g of food was provided per day, and drinking was allowed freely. Each rabbit was numbered with a 5 digit tag and each solution was labeled A, B, C respectively. Animals were sacrificed up to 3 weeks post-operatively using pentobarbital 50 mg administered intravenously, and one observer unaware of the nature of the treatment was described by Blauer et al. (1988,
Fertility & Sterility, 49: 144-49) was used to improve intraperitoneal adhesions.

[0046]

[Table 1]

The thin film adhesion was defined as an adhesion that can be easily broken. Dense adhesions were defined as adhesions that did not easily separate.

Statistical analysis was performed using STATA. The difference in adhesion score between groups was α = 0.05
Kruskall-Walli for equivalence of populations with (p = 0.02)
Determined using the s-test. Then two-sample Wilcoxo with α = 0.05
Each group was analyzed relative to the other groups using the n-rank sum analysis.

The LM609 treated group scores were similar to those seen in the sham operated group (p = 0.11), demonstrating that this treatment was effective in reducing adhesion formation. From the results, saline (p = 0.05) or anti-beta1 integrin (p = 0.0)
The control treatment of 3) was less effective when compared to the sham operated group.

[0050]

[Table 2]

The invention fully described in many of its embodiments and claimed herein can be made and executed in accordance with the teachings herein without undue experimentation. The compositions and methods of the present invention have been described by the examples above,
It will be apparent that many changes and modifications can be applied to the compositions and methods described herein without departing from the concept, spirit and scope of the invention.

─────────────────────────────────────────────────── ─── Continued front page    (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, I T, LU, MC, NL, PT, SE), OA (BF, BJ , CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, K E, LS, MW, MZ, SD, SL, SZ, TZ, UG , ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, BZ, C A, CH, CN, CR, CU, CZ, DE, DK, DM , DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, K E, KG, KP, KR, KZ, LC, LK, LR, LS , LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, R U, SD, SE, SG, SI, SK, SL, TJ, TM , TR, TT, TZ, UA, UG, UZ, VN, YU, ZA, ZW (72) Inventor Chieretsuyu, David A.             California, United States, 92024,             Encinitas, Lone Hill Rain             3277 (72) Inventor Letsy, Bruce             United States, North Carolina             27278-8802, Hillsborough, Pleasant             To Green Road 520 F-term (reference) 4C076 AA09 AA11 AA22 AA24 CC19                       EE42                 4C084 AA01 AA02 AA16 DC50 MA05                       MA13 MA17 MA55 MA56 MA66                       NA14 ZA89                 4C085 AA14 BB11 BB41 BB44 CC23                       GG01 GG02 GG04 GG05 GG06

Claims (35)

[Claims] 1. A method of inhibiting adhesion formation, comprising the step of inhibiting the binding of alphaVbeta3 integrin to extracellular matrix proteins in a mammalian body. 2. A method of inhibiting post-surgical adhesion formation comprising administering to a surgical patient an adhesion-inhibiting amount of an antagonist molecule that blocks the binding site of alpha V beta 3 integrin for binding to extracellular matrix proteins. . 3. The method according to claim 2, wherein the extracellular matrix protein is fibronectin. 4. The method according to claim 3, wherein the antagonist molecule has an antigen-binding site of the monoclonal antibody LM609. 5. The method of claim 3, wherein the antagonist molecule is the monoclonal antibody LM609. 6. The method of claim 3, wherein the antagonist molecule is the antigen binding portion of monoclonal antibody LM609 selected from the group consisting of Fab, Fab2, Fv, and mixtures thereof. 7. The method of claim 2, wherein the antagonist molecule is administered in a physiologically compatible solution at a concentration of at least about 50 milligrams per milliliter. 8. The method of claim 2, wherein the antagonist molecule is a monoclonal antibody. 9. The method of claim 2, wherein the antagonist molecule is a peptide. 10. The method of claim 2, wherein the antagonist molecule is a peptidomimetic. 11. The method of claim 2, wherein the antagonist molecule is a protein. 12. The method of claim 2, wherein the antagonist molecule is administered with an absorbable solid carrier. 13. The method of claim 2, wherein the antagonist molecule is administered with an absorbable gelatin carrier. 14. The method of claim 2, wherein the antagonist molecule is administered with a paste as a carrier. 15. The method of claim 2, wherein the antagonist molecule is administered intraperitoneally. 16. The method of claim 2, wherein the antagonist molecule is administered subcutaneously. 17. The method of claim 2, wherein the antagonist molecule is administered intravenously. 18. A composition suitable for improving adhesion formation comprising an antagonist molecule, wherein the antagonist molecule prevents the binding site of the alphaVbeta3 integrin from binding to extracellular matrix proteins. And a physiologically acceptable absorbable solid carrier vehicle that can be applied to the composition. 19. The composition according to claim 18, wherein the extracellular matrix protein is fibronectin. 20. The composition of claim 19, wherein the antagonist molecule has the antigen binding site of monoclonal antibody LM609. 21. The composition of claim 19, wherein the antagonist molecule is mouse monoclonal antibody LM609. 22. The composition of claim 19, wherein the antagonist molecule is the humanized monoclonal antibody LM609. 23. The composition of claim 19, wherein said molecule is the antigen binding site of monoclonal antibody LM609 selected from the group consisting of Fab, Fab2, Fv, and mixtures thereof. 24. The composition of claim 18, wherein the antagonist molecule is a monoclonal antibody. 25. The composition of claim 24, wherein the antagonist molecule is the antigen binding segment of a monoclonal antibody. 26. The composition of claim 18, wherein the absorbable solid carrier is gelatin. 27. The composition according to claim 18, wherein the antagonist molecule is a protein. 28. The composition of claim 18, wherein the antagonist molecule is a peptide. 29. The composition of claim 18, wherein the antagonist molecule is a peptidomimetic. 30. An alternative, physiologically acceptable carrier for an antagonist molecule that is suitable for improving post-surgical adhesions and blocks the binding of the alphaVbeta3 integrin binding site to extracellular matrix proteins. A packaged dosage form of a composition comprising together a dosage form further comprising a label indicating that the packaged inhibitor composition can be used to inhibit adhesion formation. 31. The packaged dosage form of claim 30, wherein the extracellular matrix protein is fibronectin. 32. The packaged dosage form of claim 31, wherein said antagonist molecule carries the antigen binding site of monoclonal antibody LM609. 33. The antagonist molecule is a mouse monoclonal antibody LM609.
32. The packaged dosage form of claim 31, which is: 34. The antagonist molecule is a humanized monoclonal antibody LM609.
32. The packaged dosage form of claim 31, which is: 35. The packaged dosage form of claim 31, wherein the antagonist molecule is the antigen binding site of monoclonal antibody LM609 selected from the group consisting of Fab, Fab2, Fv, and mixtures thereof. .