MXPA00001013A - Collagen type i and type iii adhesive compositions - Google Patents

Abstract

Polymerized type I and/or III collagen based compositions for medical use as adhesives and sealants and preparation thereof are described. Prior to polymerization, the collagen monomers are prepared recombinantly whereby chemical modifications of the collagen are not needed to form such monomers. The type I and/or III collagen compositions are useful as medical adhesives for bonding soft tissues or in a sealant film for a variety of medical uses. In a further aspect of the present invention, the polymerized type I and/or III collagen composition includes agents which induce wound healing or provide for additional beneficial characteristics desired in a tissue adhesive and sealant.

Description

'- ADHESIVE COMPOSITIONS OF TYPE I AND TYPE III COLLAGEN 1. FIELD OF THE INVENTION The present invention is directed to compositions based on collagen type I and / or type III recombinant, polymerized and combinations thereof for medical use as adhesives and sealants and the preparation of such compositions. Recombinant type I and type III collagen compositions are useful as medical adhesives for bonding soft tissues or in a sealing film for a variety of medical uses, including devices for closing wounds and tendon shells to prevent adhesion formation after surgical procedures. In a further aspect of the present invention, the collagen composition of type I and polymerized type III includes • agents, which induce wound healing or provide additional beneficial characteristics desired in a tissue adhesive and sealant. 20 2. BACKGROUND OF THE INVENTION Mechanical, chemical, synthetic and aatological adhesion techniques. The ability to join biological tissues is a goal of biomedical researchers. The 25 attempts to provide the desired membership through * < and* Mechanical adhesion has proven to be neither convenient nor permanent (Buonocore, M., Adhesion in Biological Systems, R. S. Manly, ed., Academy Press, New York, 1970, Cap, 15.). For example, conventional methods of choice for closing soft tissue incisions after surgery, damage and the like have been sutures and staples. These techniques and methods, however, are limited, for example, by the incompatibility of tissue with sutures or staples, which can cause pain and difficulty in treating fistulas, granulomas and neuromas. Sutures and staples may also tend to cut through weak or poorly vascularized parenchymal tissue. The sutures also leave behind a tract, which can allow the leakage of 15 fluids and organisms. The needle for any suture is larger than the thread attached to it. This produces the problem that the needle tract is larger than that which can be filled by the thread. In addition, they are limits imposed by the manual dexterity and view required of the surgeon and the excessive amount of time that is required for the use of sutures or staples in microsurgeries. Finally, even when properly applied, the joints in the spaces between the staples or sutures may be inherently weak or may weaken naturally over time and leak. Several researchers have worked on laser wound closure (White et al., 1986, White, 5 J.V., 1989, Oz and Bass et al., 1989, White et al., 1987). The first contributions focused on the welding of tissues using lasers of different wavelengths applied directly to the edges of the wound. Investigating the microstructural bases of After the tissue fusion thus produced, Schober and his collaborators proposed that a "homogenizing change in collagen with interdigitation of altered individual fibrils" occurred (Shober et al., 1986). These researchers, like others, proposed that The concentrated heating of the collagen fibrils above a threshold level allowed their cross-linking CGoosey et al., 1980; Chacón et al., 1988; Tanzer, M.L., 1973). Unfortunately, the heat necessary to allow this reaction to occur causes thermal damage collateral. Even a slight distortion, for example in ocular tissue, can have functional consequences. Also, in the case of a laser welding failure, the edges of the fabrics can be damaged by the original treatment and can not be re-exposed to the laser energy (Oz, 1990). < «£ - One more work tried to increase or improve the cross-linking caused by heat by placing a dye on the wound. It was reported that the equalization of the absorbance of the dye with the wavelength of the laser, allowed to achieve an adhesive effect with a laser of less power and less collateral thermal damage (Chuck et al., 1989; Foote, C.S., 1976; Oz M.C. and Chuck et al., 1989). The coupling of the dye with a protein was also investigated to create a "welder" of tissue. The protein of choice has been fibrinogen, and in particular autologous fibrinogen to avoid the problems of transfer of viral diseases through the use of donor blood components. In previous applications, fibrinogen has been obtained as a whole blood fraction. This is not pure fibrinogen, but it also contains other blood elements, such as coagulation factors. The application of such a protein-dye mixture in several animal models proved to be an improvement to the dye alone (Oz et al., 1990; Moazami et al., 1990). Unfortunately, the human application was delayed due to the need to isolate the necessary protein (fibrinogen) from the patient before the procedure to avoid the risks of infection of the donor plasma. Working with albumin found that this is a substitute unsatisfactory, since it does not produce welds of comparable strength. Comparisons of protein-dye closures versus sutures have found that the protein-dye group produces less inflammatory response, as a result of increased collagen production, higher peak peak stress at break and better cosmetic effects (Wider et al., 1991 ). The ophthalmic application of such a tissue welder has included the sealing of conjunctival blisters (Weisz, et al., 1989), sclerostomy (Odrich et al., 1989), closure of rhinotomies (Wolf et al., 1989), and thermokeratoplasty ( Wapner et al., 1990) using similar mixtures. Due to the deficiencies and limitations of these mechanical means, whether sutures, staples or the most recently applied laser techniques, much attention has been devoted to developing synthetic polymers, for example, cyanoacrylates, as biomedical adhesives. However, it has been observed that these plastic materials induce inflammatory tissue reaction. In addition, the ability of these materials to establish a permanent bond under physiological conditions has not yet been fully achieved. The known toxicity associated with synthetic adhesives has led to investigations into the development of biologically derived adhesives as binding materials. Among such adhesives, cements • Fibrin-based have gained considerable attention. (See, for example, Epstein, G.H. et al., Ann, Otol.

Rhinol. Laryngol, 95; 40-45 (1986); Kram, H. B. et al. Arch. Surg. 119: 1309-1311 (1984); Scheele, J. et al. Surgery 95; 6-12 (January 1984); and Siedentop, K.H. et al. Laryngoscooe 93: 1310-1313 (1983) for the general discussion of fibrin adhesives). The adhesives commercial fibrin tissue are derived from human plasma and consequently possess potential health risks, such as adverse immunogenic reactions and the transmission of infectious agents, for example, the hepatitis B virus. In addition, the binding force imparted by such adhesives is relatively weak compared to collagen adhesives (See De Toledo, A.R, et al., Assoc. For Res, in Vision and Ophthalmology, Annual Meeting Abstract, Vol. 31, 317 (1990)). Consequently, there is a need for biologically compatible, effective and safe tissue adhesives for bio-medical applications. More recently, combined products have been devised to be used as a tissue adhesive. For example, Staindl (Ann. Otol (1979) 88: 413-418) describes the use of a combination of three substances prepared separately, the cryoprecipitate of human fibrinogen, thrombin in the presence of calcium ion, and concentrate of Factor XIII, to obtain a cement that was applied in applications of skin grafting, myringoplasty, 5 repair of dural defects, haemostasis after tonsillectomy, and tracheoplasty. In this same time interval, Immuno-AG, Vienna, Austria, began to produce and market a "fibrin seal" system where one component contains fibrinogen highly concentrated human, Factor XIII, and other human plasma proteins, prepared from whole blood, and the other component supplies thrombin and calcium ions. The two components are added together in the presence of a fibrinolysis inhibitor. After of the application, the processes of coagulation and cross-linking of fibrin occur. Eventually, the seal • can be used in the process of wound healing or trauma that accompanies tissue reconstruction. Redi, H., et al., "Biomaterials 1980", Winter, G.D., et al., Eds. (1982), John Wiley & Sons, Ltd., on page 669-675, describes the development of an applicator device for this system, which mixes and applies the two components of the system simultaneously. These combined systems and their use have been widely described: Seelich, T., J Head and w i * - * J «go -« * *% & Neck Pathol (1982) 3: 65-69; O'Connor, A, F., et al., Otolaryngol Head Neck Surg (1982) 90: 347-348; Marquet, J., J • Head and Neck Pathol (1982) 3: 71-72; Thorson, GK, et al., J Surg Oncol (1983) 24: 221-223, McCarthy, PM, et al., May 5 Clin Pros (1987) 62: 317-319, reported the addition of barium ion to this system of fibrin cement in the treatment of a hemorrhagic duodenal sinus to facilitate the monitoring of survival. See also Portmann M., J Head and Neck Pathol (1982) 3:96; Pañis, R., ibid., 94-95. Recently efforts have also been focused on methods that seek to avoid health concerns arising from the use of blood plasma derived products in commercially available tissue adhesive products and systems. To this end, 15 attempts have been made with varying degrees of success by isolating an autologous counterpart of the fibrinogen-containing component. For example, see Feldman, M.C., et al., Arch Otolaryngol-Head and Neck Surg (1988) 114: 182-185; Feldman, M. C, et al., Arch Ophthalmol (1987) 105; 963-967; Feldman, 20 M. C, et al., M J Otolog (1988) 9: 302-305; Siberstem, L.E., et al., Transfusion (1988) 28: 319-321. The use of autologous fibrinogen preparations also has obvious limitations. you # '* # * * - # 3 Collagen as a bio aterial. Collagen, the principal connective tissue protein in animals, possesses numerous characteristics not observed in synthetic polymers. The characteristics of collagen 5 frequently cited include its good compatibility with living tissue, promotion of cell growth, and absorption and assimilation of implants (Shimizu, R. et al., Biomat.Med. Dev. Art. Org., 5 (1). ): 49-66 F (1977)). Various applications of this material are being tested, for example, as artificial kidney dialysis membranes (Sterzel, KH et al., A eri, Soc. Artif. Int. Organs 17: 293 (1971)), artificial-horneatal ( Rubin, AL et al., Nature 230: 120 (1971) and U.S. Patent No. 4,581,030), vitreous body (Dunn, M. et 15 al. Amer. Soc. Artif. Int. Organs 17: 421 (1971)), skin and artificial blood vessels (Krajicek, M. et al., J. F Surg, Res.4,290 (1964)), as hemostatic agents (US Patent No. 4,215,200), soft contact lenses (US Patent Nos. 4,264,155; 4,264,493). 4,349,470, 4,388,428, 4,452,925 and 4,650,616) and in surgery (Chvapil, M. et al., Int.Rev. Conn. Tiss, Res. 6: 1-61 (1973)). Natural collagen fibers, however, are basically soluble in mature tissues due to covalent intermolecular cross-links that ao * _ They convert collagen into an infinite reticulated network. The < ? dispersion and solubilization "The native collagen can be achieved by treatment with several proteolytic enzymes, which break the intermolecular bonds and remove immunogenic non-helical extreme regions without affecting the rigid, basic triple helix structure that imparts the desired characteristics of the collagen. {see also, US Patents Nos. 3,934,852, 3,121,049, 10 3,131,130, 3,314,861, 3,530,037, 3,949,073, 4,233,360 and 4,488,911 for general methods to prepare purified soluble collagen.) Several methods and materials have been proposed to modify collagen to make it more suitable as a biomedical adhesive (See, for example, De Toledo, AR et al., Assoc. For Res. in Vision and Ophthalmology, Annual Meeting Abstract, Vol. 31, 317 (1990); Lloyd et al., "Covalent Bonding of Collagen and Acrylic Polymers ", American Chemical Society Symposium on Biomedical and 20 Dental Applications of Polymers, Pol ymer Science and Technology, Vol. 14, Plenum Press (Gebelein and Koblitz eds.), New York, 1980, p. 59-84; Shmizu et al., Biomat. Med. Dev. Art. Org., 5 (1): 49-66 (1977); and Shimizu et al., Biomat. Med. Dev. Art. Org., 6 (4): 375-391 (1978), 25 for general discussion of collagen and polymers S_s_ synthetic). In many cases, the above modified collagen-based adhesives suffer from several • deficiencies, which include (1) cross-linking / polymerization reactions that generate exothermic heat, (2) extended reaction times and (3) reactions that do not operate in the presence of oxygen and the physiological pH ranges (Lee ML et al., Adhesion in Biological Systems, RS Manly, ed., Academic Press, New York, 1970 , Chapter 17). In addition, many of the adhesives based on The above modified collagen contains toxic materials, rendering them unsuitable for biomedical uses (see, for example, Buonocore, M. G. (-1970) and US Pat. No. 3,453,222). Additionally, the use of adhesives based on collagen also present immunological concerns since such adhesives have been • derived from animal sources and typically bovine sources. Studies regarding the use of such collagens as injectable devices have reported minor inflammatory responses. More recently, attention has been focused on potential aspects with respect to the transmission of disorders to humans related to bovine spongiform encephalopathy ("mad cow disease") especially in Europe, to limit materials of bovine origin. *""or Notwithstanding these deficiencies, certain collagen-based adhesives have been reported, which have been reported to have strength and appropriate adhesive utility in many medical applications, particularly those involving the soft tissues. U.S. Patent No. 5,219,895). These reports identify the use of collagen-based adhesives of type I and type II; where the collagen of purified types I and II was chemically modified to form monomers, which are soluble at physiological conditions and then polymerized to form a composition having adhesive and sealing properties. The reports are limited to collagen-based adhesives, which are composed of collagen derived from natural sources; and consequently, they represent a mixture of collagen. For example, type I collagen, as isolated from natural sources, is typically comprised of approximately 10-20% type III collagen and other collagens, depending on the tissue source used, and 90-80% collagen of the type I. The reports, moreover, do not refer to type III collagen, the unexpected haemostatic characteristics of type III collagen or the use of recombinant collagens so that the chemical modification of the first step can be avoided. -_. 13" 3. BRIEF DESCRIPTION OF THE INVENTION A biologically compatible type III and / or type I collagen product with sealant and adhesive properties can be formed using collagen monomers type III and / or type I recombinantly derived, soluble; wherein the monomers are polymerized to form a type III and / or type I collagen composition having adhesive and sealant properties. Preferably, the collagen is human and is derived using recombinant technology. Collagen type III was selected for its unexpectedly superior haemostatic characteristics, compared to other types of collagen. Collagen type I was selected for its structural characteristics. The polymerization reaction can be initiated with an appropriate polymerization initiator such as a chemical oxidant, ultraviolet irradiation, a suitable oxidant enzyme or atmospheric oxygen. In order to optimize the sealant and adhesive properties of the recombinant collagen product using the structural stability of the product, as well as the hemostatic characteristics of the product, the product is preferably comprised of a collagen composition of type I and type III recombinant, pure. The ratio of pure recombinant type III collagen to pure recombinant type I collagen is about 30% and greater than type III collagen to about 70% or less of type I collagen. More preferably, the collagen type ratio Pure recombinant III, to the pure recombinant type I collagen, is from about 30% to about 50% of type III collagen up to about 70% to about 50% of type I collagen. More preferably, the collagen ratio of the Pure recombinant type III, to pure recombinant type I collagen is from about 30% to about 40% of type III collagen up to about 70% to about 60% of type I collagen. The aim of the invention is to provide a collagen tissue sealer of pure recombinant type III, a pure recombinant type I collagen tissue sealer or a tissue collagen sealer of the ti po I and type III pure recombinant free of other types of collagen of the type Input test cases, which has the following characteristics and capabilities: (i) Hemostasis. The sealant acts as a hemostatic barrier and reduces the risk of leakage of serum, lymph and liquor. Since type III collagen possesses inherent hemostatic properties, its use in a Haemostatic device provides an improvement over known fibrin sealants. Collagen type I also has some hemostatic properties. (ii) Paste. Due to its adhesive properties, the sealant automatically connects the tissues forming a strong bond between them and adapts non-uniform wound surfaces. This sticking effect is increased by a combination of agents, as described below, and type III collagen and / or type I collagen. (Iii) Wound healing. The sealant promotes the growth of fibroblasts, which in combination with efficient haemostasis and adhesion between wounded surfaces provides an improved healing process. The use of the compositions according to the invention as a non-stick / wound healing composition is expected to result in a normal (regenerative) tissue instead of scar tissue, ie optimal wound healing. In addition, such compositions also reduce the inflammatory response. Accordingly, the object of the present invention is to provide collagen compositions of type III and / or type I polymerized as effective, safe biological adhesives, with strong properties. adhesives for biomedical applications, particularly those involving soft tissues. More specifically, the present invention is directed to compositions useful for sealing punctures or incisions in internal organs, the dermis and large blood vessels. The polymerized materials can assume a number of sizes and shapes consistent with their intended biomedical applications, which include use in ophthalmology, plastic surgery, orthopedics and cardiology. In another object of the invention, the collagen composition of type III and / or type I is further comprised of agents which confer additional desirable characteristics for a sealant or adhesive. For example, fibroma, fibrinogen, thrombin, calcium ion, factor XIII can be included in the composition to better effect the formation of a three-dimensional network of polymerized collagen. In yet another object of the invention, the recombinant type III collagen composition incorporates a drug that has a wound healing ability. In one embodiment, the drug is a factor of connective tissue growth and is incorporated into the composition for effect a slow release of the drug to the wound, 4. DETAILED DESCRIPTION OF THE INVENTION 4.1 Definitions 5 As used herein, the term "biologically compatible" refers to type III collagen and / or modified type I recombinant according to the present invention (ie, a recombinant collagen type III polymerized product) which is incorporated or implanted in or placed • 10 adjacent to the biological tissue of a subject and more particularly, does not deteriorate appreciably over time or induces an immune response or harmful tissue reactions after such incorporation or implantation or placement. As used here, the term "collagen from Pure recombinant type I "refers to collagen of human type I manufactured by recombinant techniques, which is substantially free of other types of collagen, the term excludes type I collagen isolated from natural sources. the term "pure recombinant type III collagen" refers to human type I collagen manufactured by recombinant techniques, which is substantially free of other types of collagen.The term excludes collagen type III isolated from natural sources. and III. The type of collagen useful in forming the collagen product 5 biologically compatible with the adhesive and hemostatic properties of this invention is collagen type I and III recombinant. The monomeric soluble type I and III collagen is obtained by recombining processes, including the processes that involve production of type III collagen in transgenic animals. Recombinant processes are set forth in U.S. Patent No. 5,593,859, which is incorporated herein by reference. Preferably, collagen of type I or III will be manufactured in a manner Recombinant by culturing a cell that has been transfected with at least one gene encoding the polypeptide comprising collagen type I or II and the genes encoding the subunits and β of the post-translational polymeric enzyme 4-hydroxyslase and purifying the collagen monomer resulting therefrom. Preferably, the monomeric soluble type I and III collagen material exhibits a viscous consistency and several degrees of transparency and clarity.

Polymerization of Collagen Monomers of Type I and III. The collagen solution of type I and Recombinant F III can then be subjected to polymerization or crosslinking conditions to produce the polymerized collagen composition of the present invention. The polymerization can be carried out using irradiation, for example, UV, gamma, or fluorescent light. UV irradiation can be carried out in F the short wavelength range using a standard 254 nm source or using UV laser sources. With a standard 254 nm source, 4-12 watts, the polymerization occurs from 10 to 40 minutes, preferably from 20 to 30 minutes, at an exposure distance of 2.5-10 cm, preferably from 2.5 to 4. cm away. Excessive exposure to UV will begin to depolymerize the collagen polymers. Polymerization using gamma irradiation can be performed using 0.5 to 2.5 Mrads. Excessive exposure to gamma irradiation will also depolymerize the collagen polymers. Polymerization in the presence of oxygen can be accomplished by adding an initiator to the fluid before exposure. Non-limiting examples of initiators include sodium persulfate, sodium thiosulfate, chloride tetrahydrate ferrous, sodium bisulfite and oxidative enzymes such 4. '* ,-* - twenty* • such as peroxidase or catechol oxidase. When Ae employ initiators, polymerization occurs in 30 seconds to 5 minutes, usually 1 to 3 minutes. The polymerizing agent is preferably UV irradiation. However, the polymerization or crosslinking of the monomeric substituents can be carried out simply by exposing the material to atmospheric oxygen, although the polymerization rate is appreciably slower than in the case of UV irradiation or chemical agents. Other agents may also be useful in the polymerization process. For example, to improve the cohesion resistance of the adhesives formed from the compositions of this invention, dysfunctional monomeric crosslinking agents can be added to the monomer compositions of this invention to effect polymerization. Such crosslinking agents are known in the art, such as, for example, US Patent No. 3,940,362 (Overhults), which is incorporated herein by reference. 4. 3 Collagen Compositions of Type I and III The compositions of the present invention are comprised of collagen type I and III polimepzado, where the composition is manufactured by an i-fVJ process comprising the steps of: (1) production of collagen monomers from A * jffte * I and III by the recombinant method described above; , and (2) polymerization of such monomers. For the purposes of optimizing the sealant and adhesive properties of the recombinant collagen product by optimizing the structural stability of the product, as well as the hemostatic characteristics of the product, the product is preferably comprised of a combination of collagen type I and type III pure recombinant. The ratio of pure recombinant type III collagen to pure recombinant type I collagen is about 30% and greater of type III collagen to about 70% or less of type I collagen. More preferably, the ratio of collagen of the pure recombinant type III to pure recombinant type I collagen is from about 30% to about 50% of type III collagen up to about 70% to about 50% of type I collagen. More preferably, the collagen ratio of the Recombinant type III to recombinant type I collagen is from about 30% to about 40% type III collagen about 70% to about 60% type I collagen. - fee \ ß £ The compositions of the present invention may also be comprised of other agents which are useful for sticking or sealing fabrics. For example, in addition to the recombinant type I and / or type III collagen protein, the composition will preferably comprise Factor XIII and / or fibrin / fibrinogen / fibronectin and / or plasminogen. Advantageously, the composition will also include coagulant enzyme, i.e., thrombin, especially in combination with divalent calcium, such as the calcium chloride. The concentration of calcium chloride will then vary, for example, between 40 mM to 0.2 M depending on the specific purpose - of the tissue adhesive composition, high concentrations of calcium chloride inhibit the growth of the fibroblasts and therefore, are preferred for anti-adhesion applications (together with the absence of fibronectm, which stimulates the growth of fibroblasts). In addition, it may be valuable to include a fibrinolysis inhibitor, such as a plasmin inhibitor, for example, aprotinin, applotinin, alpha-2-antiplasmin, alpha-2-macroglobulin, alpha-1-antitrypsin, epsilon-aminocaproic or tranexamic acid, or an inhibitor of the plasmin activator, for example PAI-1 or PAI-2. Although the proportions of previously known ingredients in adhesive compositions tissue of the invention can be selected with the guide v? "> * 2« 3 of the prior art compositions, the necessary amount of polymer to increase the viscosity; they can easily be determined by one skilled in the art depending on the particular polymer and the intended form of use. Thus, if the concentration and / or molecular weight of the viscosity-increasing polymer is too low, the increase in viscosity will be insufficient, and too high a concentration and / or molecular weight will inhibit fibrin polymerization and adhesion to the fibrin. tissue. By increasing the thrombin concentration, the polymerization of the composition of the present invention can be accelerated with a consequent influence on the time until the cement or glue hardens. At low concentrations of thrombin, for example, the fibrin in the composition will remain more or less fluid for several rt) inutes after application. An additional beneficial effect of increasing the viscosity with a polymer that increases the viscosity according to the invention is therefore the possibility of using lower concentrations of thrombin, which is required in situations where the parts to be sealed require a subsequent adaptation even on non-horizontal surfaces. In the same way, the compositions of the present the agents described herein, are a fusion protein where collagen of the? -F4 type III type and, for example, fibrin, combine to form a molecule. Such fusion proteins can be manufactured according to the recombinant techniques described herein. In a further embodiment of the invention, the composition of the present invention includes agents useful in wound healing, either by inducing or promoting tissue Aormation, or alternatively, limiting the formation of fibrotic adhesions. Such agents include antibiotics or growth factor, such as connective tissue growth factor, which is described in, for example, U.S. Patent Nos. 5,408,040 and 5,585,270, the references are incorporated herein by reference. 4. 4 Fields of Use The polymerized type III and / or type I collagen product can be useful for producing sealing systems and mechanical adhesives. Tissue Adhesive Systems. * Fields of application include among others: surgery of the ear, nose and throat, general surgery, dentistry, neurosurgery, plastic surgery, thorax and vascular surgery, abdominal surgery, orthopedics, surgery accidents, gynecology, urology and ophthalmology. The collagen sealants of this invention have also been used for the local application of drugs, such as antibiotics, growth factors and cytostatic agents Sealing Films In one aspect of the invention, the polymerized collagen products can be made in the form of a sealing film.A collagen-based film will be flexible and elastic with the consistency and feel of a plastic film, and in addition the film should exhibit a high biological compatibility.The use of sealing films include: Prevention of adhesion formation after tendon surgery (ie use as a wrap around the tendons), use as a synthetic tympanic membrane, substitute facial tissue and dressing component of bandages and wounds. Potential use of sealing films include: treatment of corneal abrasions, wound closure, coating of catheters and instruments, use as a material to prevent the formation of adhesion in tissues other than tendons (eg, peritoneal cavity). Additional embodiments of the present invention include sealant and adhesive formulations, "i-y *. which can be used as specific systems for the release of numerous drugs and pharmaceutical compositions, including growth factors, antibiotics, and other biologically beneficial compounds. Such materials can be added to the adhesive or collagen sealant to promote cell migration, cell adhesion and wound healing. Angioplasty and angiography. Angiography is a diagnostic procedure by which a dye is injected into the artery, preferably the femoral artery, to detect the presence or absence of coronary disease. Angioplasty, also known as PCTA, is a therapeutic procedure which involves inflating a balloon into an artery, such as the coronary artery, for the purpose of relieving arterial blockages. After a puncture in the femoral artery, the balloon catheter is inserted through the femoral artery and navigates through the coronary artery blocked by atherosclerosis (plaque). Once in position, the balloon is inflated and deflated several times in an effort to open the artery by pushing the fatty material against the walls of the vessel, allowing blood to circulate to the affected regions of the heart muscle. Several s of balloon catheters are ¡? ~ *, 27 commonly used in angiosplasty and angiography, including catheters on wire, which run F over an independent guidewire to the site of the disease; 2) fi bre wire catheters, which combine a balloon catheter with a guide wire in a device; 3) quick exchange or single-operator exchange catheters, which run over wire catheters and can be exchanged more conveniently than catheters running on standard wire; and 4) perfusion catheters, which allow blood flow during the procedure. A rotating tip catheter removes the accumulated plaque on the arterial walls. These devices use a technique called differential cutting. The calcified material is converted into microscopic particles without damaging the artery due to the elastic nature of the arterial walls. Angioplasty is a more invasive and complicated procedure than angiography, since it requires the insertion of a larger sheath than that used in angiography. The sheath is used as a vehicle to introduce the catheter into the artery. Additionally, angioplasty also requires the use of blood thinners, such as heparin, to prevent coagulation during and after the surgical procedure. The anticoagulant agent prevents the body's natural sealing / coagulation mechanism and, thus, sealing punctures requires a significant period of time. According to the present invention, after removing the catheter or other invasive devices from the artery, an adhesive applicator may optionally be inserted into the sheath and placed in a position near or in contact with the puncture of the artery. During the procedure, manual or mechanical pressure is applied to reduce blood flow at the puncture site. If possible, remove excess blood / fluid from the puncture site. Subsequently, recombinant type III and / or type I collagen monomer of the present invention can be applied to the puncture on the external surface of the artery and / or within the puncture channel. The monomer is then polymerized and / or crosslinked by the techniques described herein, for example, UV irradiation, so that the polymerization takes place within 0 to 300 seconds, preferably 0 to 120 seconds, more preferably 0 to 30 seconds. seconds, and even more preferably 3 to 10 seconds. Applying the composition of collagen monomer on the outside of the artery, the incidence of embolism (blockage of the artery or system 29 circulatory) is virtually eliminated. Alternatively, a type III collagen and / or polymerized type I collagen can be used, the polymerization step can be avoided. Due to the bond strength of the adhesive of the present invention, only small amounts of adhesive are required to seal the perforated artery. In addition, because the surgical adhesive according to the present invention can polymerize almost immediately, the adhesive can polymerize on the surface and / or along the puncture channel of the artery without penetrating into the artery. Consequently, large pieces or particles of material will not enter the circulatory system, thereby substantially reducing the risk of embolism. Due to the fast and strong bonding of the preferred adhesives of the invention, the patient needs to be immobilized only for a minimum period of time. 4. 5 Administration Formulations. The tissue treatment composition of the present invention can be presented in the same type of preparations of the fibrin sealants of the prior art. The components can be provided in the form of a frozen solution or as lyophilized powders, to be diluted before 30 minutes. used with the appropriate aqueous solutions, for example containing aprotinin and calcium ions, respectively. With respect to the compositions of the present invention comprising pharmaceutical agents, such as an antibiotic, a growth factor, etc., by incorporating the agent into the tissue adhesive to be enclosed in the collagen network formed after the application of the tissue adhesive Therefore, it will be ensured that the drug is maintained at the site of application while being released in a controlled manner from the composition, for example when used as eye drops, a preparation for wound healing, etc. As mentioned above, the pharmaceutically active substance to be released from the tissue adhesive composition of the present invention may be the viscosity-increasing polymer itself or a substance coupled thereto. As a specific example of such viscosity-increasing polymer that satisfies the requirement to increase viscosity, as well as having therapeutic and pharmaceutical utility, and for which one may wish to sustain bioavailability, is hyaluronic acid and its salts and derivatives thereof. They are easily soluble in water and, as mentioned above, have an extremely short shelf life. The tissue treatment composition of this invention thus constitutes an advantageous slow release preparation for proteoglycans such as hyaluronic acid and its salts and derivatives, and considerably increases the bioavailability thereof. Notably, the compositions of the present invention are not restricted to the adhesive properties, but non-adhesive compositions are also included, especially when the composition is intended to serve primarily to heal wounds. The latter compositions may in particular include non-adhesive proteins such as albumin and / or growth factors. Substantially non-adhesive compositions can also be obtained when the polymer part of the composition inhibits the aclhesive properties of the protein part. In this context it should be emphasized that the invention comprises adhesive and substantially adhesive compositions, although this, for reasons of simplicity, has often been referred to as an "adhesive" in this specification. Application of the Compositions. The compositions of the present invention can be applied using a variety of dispensing devices. For example, the surgical adhesive can '32 be applied using the devices set forth in U.S. Patent Nos. 4,900,303 (Lamelson) and 5,372,585 (Tiesenbrun) which simultaneously verify the application process through an optical observation system. The composition of the present invention can also be applied by the devices set forth in U.S. Patent No. 5,129,882 (Weldon et al). The subject matter of these patents is incorporated herein by reference. The composition according to the present invention can also be applied in conjunction with other sealing means. For example, the adhesive can be applied to the puncture sites that have been closed using sutures or surgical caps, such as in the sealing of a puncture or incision in internal organs, for example, the liver, gallbladder, intestines, stomach , kidney, heart, urinary bladder, ureter, lung, esophagus and the like. The adhesive in this case will provide a complete seal, thereby reducing the risk of leakage of blood from the organ or vessel, for example, leakage of the hepatic puncture sites. The surgical adhesive of the present invention can be further used in conjunction with other sealing means, such as plugs, and the like. Such techniques are disclosed in the Patents ^, - * • »£ i 33 US Nos. 4,852,568 (Kensey), 4,890,612 (Kensey), 5,053,046 (Janese), 5,061,272 (Kensey), 5,108,421 (Fowler), 4,832,688 (Sagae et al), 5,192,300 (Fowler), 5,222,974 (Kensey et al.), 5,275,616 (Kensey). Fowler), 5 5,282,827 (Kensey et al.), 5,292,332 (Lee), 5,324,306 (Makower et al.), 5,370,660 (Weinstein et al.), And 5,021,059 (Kensey et al.). The subject matter of these patents is incorporated herein by reference. Notably, the compositions of this invention can be used to join two surfaces by applying the particular composition to at least one of the surfaces. Depending on the particular requirements of the user, the adhesive compositions of this invention can be applied by known means such as a glass stirrer, sterile brush or medicine dropper; However, in many situations, • prefers a pressurized aerosol dispensing package in which the adhesive composition is in solution with a compatible anhydrous propellant. The aerosol application of the monomers is particularly advantageous for use in haemostasis. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (20)

*!**-, 3. 4 CLAIMS Having described the invention as above, the content of the following claims is claimed as property.

1. A tissue adhesive or sealant composition, characterized in that it comprises a polymerized type III collagen, wherein the sealant adhesive composition is produced by recombinantly manufacturing pure type III collagen monomers in a cell and polymerizing the monomers with an agent.

2. The composition according to claim 1, characterized in that the composition is biologically compatible.

3. The composition according to claim 1, characterized in that the recombinant manufacture of a type III collagen monomer comprises the following steps: (a) culturing a cell, which has been transfected by at least one gene coding for a polypeptide comprising type III collagen and at least one gene encoding a polypeptide selected from the group of the a or β subunit of prolyl-4-hydroxylase; and (b) purifying type III collagen.

4. The composition according to claim 1, characterized in that the monomers are polymerized using irradiation.

5. The composition according to claim 4, characterized in that the irradiation is UV irradiation.

6. The composition according to claim 1, characterized in that the composition is further comprised of one or more agents selected from the group of fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor.

7. A process for manufacturing a sealant or tissue adhesive, characterized in that it comprises the steps of: (a) manufacturing a collagen monomer of type III by recombinant means; and (b) polymerizing collagen monomers of type III.

8. A tissue sealant adhesive composition, characterized in that it comprises a polymerized type I collagen, wherein the adhesive or sealant composition is produced by recombinantly manufacturing pure type I collagen monomers in a cell and polymerizing such monomers with an agent. • V "36

9. The composition according to claim 8, characterized in that the composition is biologically compatible.

10. The composition according to claim 8, characterized in that the recombinant manufacture of a collagen monomer of type I comprises the following steps: (a) culturing a cell, which has been transfected by at least one gene coding for a polypeptide comprising collagen of type I and at least one gene encoding a polypeptide selected from the group of the a or β subunit of prolyl-4-hydroxylase; Y (b) purifying type I collagen.

11. The composition according to claim 8, characterized in that the monomers are polimepped using irradiation.

12. The composition according to claim 11, characterized in that the irradiation is UV irradiation.

The composition according to claim 8, characterized in that the composition is further comprised of one or more agents selected from the group of fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor. 37

14. A process for manufacturing a sealant or tissue adhesive, characterized in that it comprises the steps of: (a) manufacturing a collagen monomer of type III by recombinant means; and (b) polymerizing collagen monomers of type III.

15. A tissue adhesive or sealant composition, characterized in that it comprises a pure polymerized type III collagen and a polymerized, pure type I collagen.

16. The composition according to claim 15, characterized in that the composition is biologically compatible.

The composition according to claim 15, characterized in that the ratio of collagen of pure recombinant type III to pure recombinant type I collagen is about 30% or greater of type III collagen of about 70% or less of collagen of the type I.

18. The composition according to claim 15, characterized in that the monomers are polymerized using irradiation.

19. The composition according to claim 15, characterized in that the irradiation is UV irradiation. The composition according to claim 15, characterized in that the composition is further comprised of one or more agents selected from the group of fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor. Compositions based on collagen type I and / or III polymerized for medical use as adhesives and sealants and the preparation thereof are described. Prior to polymerization, the collagen monomers are prepared recombinantly, whereby chemical modifications of the collagen to form such monomers are not necessary. Collagen compositions of type I and / or III are useful as medical devices for bonding soft tissues or in a sealing film for a variety of medical uses. In a further aspect of the present invention, the collagen composition of type I and / or III includes agents which induce wound healing or provide additional beneficial characteristics desired in a sealant and tissue adhesive.