JP2001511431A - Type I and type III collagen adhesive compositions - Google Patents

Abstract

  (57) [Summary] Polymerized type I and / or type III collagen-based compositions and their preparation for medical use as adhesives and sealants are described. Prior to polymerization, collagen monomers are prepared recombinantly, so that no chemical modification of the collagen is required for the formation of such monomers. Type I and / or III collagen compositions are useful as medical adhesives for bonding soft tissues or in sealant films for various medical applications. In yet another aspect of the invention, these polymerized type I and / or type III collagen compositions induce wound healing or provide additional advantageous properties desired in tissue adhesives or sealants. Containing reagents.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001] This application is related to US Provisional Patent Application No. 60/0, filed Jul. 28, 1997.
No. 53,872 and claims priority.

[0002] 1. FIELD OF THE INVENTION The present invention relates to polymerized recombinant type I and / or type III collagen-based compositions and their combinations 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 sealant films for various medical applications, which are useful for adhering post-surgical procedures. Wound closure device and tendon wrap that do not interfere with formation
Is included. In a further aspect of the invention, the polymerized type I and type III collagen compositions include reagents that induce wound healing or provide additional beneficial properties desired in tissue adhesives or sealants.

[0003] 2. BACKGROUND OF THE INVENTION Mechanical, Chemical, Synthetic and Self-Adhesive Techniques The ability to bind biological tissue is the purpose of biological research. Attempts to provide the desired adhesion via mechanical bonding have not proven to be convenient or durable (Bu
onocore, M .; , Adhesion in Biological Sy
stems, R.A. S. Manly ed., Academic Press, New York, 1970, Chap. 15). For example, the choice of conventional methods for closing an incision in soft tissue after surgery, injury, etc., has been suturing and stapling. However, these techniques and methods are limited, for example, by tissue incompatibility with suturing or stapling, which can cause pain and can be difficult to treat for fistula granulomas and neuromas. Sutures and stapling may also tend to cut through weak soft tissue or poorly vascularized tissue.
Sutures also leave pathways that may allow for leakage of fluids and organisms.
Any suture needle is larger than the thread passed through the needle. This causes the problem that the needle path is larger than can be filled by the thread.

In addition, limitations are ameliorated by the required dexterity and vision of the surgeon and excessive time, which require the use of sutures and stapling in microsurgery. Finally, even when applied correctly, the bond in the gap between the stapling or suture may be inherently weak or may become structurally weak over time and leak.

[0005] Several researchers have studied laser closure of wounds (White
1986; White, J. et al. V. Oz and Bass et al., 19;
89; White et al., 1987). Welded microstructures using lasers of different wavelengths (
The early contribution, focusing on welding tissue, was applied directly to the edge of the wound. By studying the microstructure of tissue fusion, Shober and co-workers have proposed that a "uniform change in collagen with altered individual fibril fitting" results (Schober et al., 1986). These investigators, as well as others, have proposed concentrated heating of collagen fibrils above a threshold level that allows their cross-linking (Goosey et al., 1980; Chacon et al., 1988;
Tanzer, M .; L. , 1973). Unfortunately, the heating required to cause this reaction causes thermal damage to the side branch. Even small distortions can be functionally significant, for example in eye tissue. Also, in the event of a laser welding failure, the edges of the tissue can be damaged by the initial procedure and cannot be re-exposed to laser energy (Oz, 1990).

[0006] Further studies have attempted to improve heat-activated cross-linking by placing dyes on the wound. Matching of dye absorption by laser wavelength reported that the adhesive effect was achieved without laser power and thermal damage to side branches (Chuck et al.,
1989; Foote, C.I. S. Oz M., 1976; C. And Chuck
1989). The association of dyes with proteins to form tissue "solders" has also been studied. The choice of protein is fibrinogen,
And especially autologous fibrinogen to avoid the problem of viral disease metastasis through the use of blood components from pooled donors. In previous applications, fibrinogen was obtained as a fraction of whole blood. It is not pure fibrinogen but also contains other blood components (eg, clotting factors). Application of such a protein-dye mixture in various animal models provides an improvement over dye alone (Oz et al., 1990; Moazami et al., 1990). Unfortunately,
Luminal application is obviated because of the need to isolate the required protein (fibrinogen) from the patient before the procedure to avoid the risk of infection from donor plasma. Studies with albumin have not been found to be satisfactory substitutes since they do not produce comparable strength bonds.

A comparison of protein-dye versus suture closure finds protein-dye groups that produce little inflammatory response, resulting in better collagen production, better average peak stress in rupture and better cosmetology (Wider et al., 1991). Ophthalmic application of such tissue brazing is to seal the zygotic blister using a similar mixture (W
eisz et al., 1989), pleurotomy (Odrich et al., 1989), retinal resection closure (Wolf et al., 1989), and heated keratoplasty (
Wapner et al., 1990).

[0008] Due to the deficiencies and limitations of these mechanical means, much attention has been paid to synthetic polymers (eg, cyano) as biomedical adhesives, whether suturing, stapling or recently applied laser technology. Acrylates).
These plastic materials, however, have been observed to cause an inflammatory tissue response. Furthermore, the ability of these materials to establish durable bonds under physiological conditions is not yet fully understood.

The known toxicity associated with synthetic adhesives is being investigated towards the development of biologically derived adhesives as binding materials. Among such adhesives, fibrin-based glues have received considerable attention (for a general discussion of fibrin adhesives, see, for example, Epstein, GH et al., An.
n. Otol. Rhinol. Laryngol. 95: 40-45 (1986
Kram, H .; B. Arch. Surg. 119: 1309-1311
(1984); Scheele, J .; Surgery 95: 6-12 (1
Jan. 984); and Siedentop, K .; H. Et al., Laryngosc
oo 93: 1310-1313 (1984)). Commercially available fibrin tissue adhesives are derived from human plasma and therefore carry potential health risks such as adverse immunogenic reactions and transmission of infectious agents (eg, hepatitis B virus). Furthermore, the bonding strength provided by such adhesives is relatively weak compared to collagen adhesives (De Toledo, AR, et al., Assoc. For Res. In Vision and Ophthalmology, Ann).
ual Meeting Abstract, Vol. 31, 317 (1990)). Accordingly, there is a need for a safe and effective biocompatible tissue adhesive for biomedical applications.

[0010] More recently, product combinations have been devised for use as tissue adhesives. See, for example, Staindl (Ann. Otol (1979) 88: 4).
13-418) are three separately prepared substances (human fibrinogen cryoprecipitate, thrombin in which calcium ions are present, and factor XIII concentrate)
Describes the use of a combination of the following to obtain an adhesive that is applied to skin graft application, eardrum formation, repair of dural defects, homeostasis after tonsillectomy, and tracheoplasty. In this simultaneous frame, the Immuno-AG, Vie
nna, Austria has begun the manufacture and sale of a two-component "fibrin-sealed" system, where one component comprises highly concentrated human fibrinogen, factor XIII, prepared from pooled blood, and Contains other human plasma proteins, and the other component supplements thrombin and calcium ions. This 2
The two components are added together in the presence of a fibrinolysis inhibitor. After applying
The process of coagulation and fibrin crosslinking occurs. Ultimately, the seal may dissolve during the healing of wounds or trauma associated with tissue reconstruction. Redl, H .; "Bi
omaterials 1980 ", Winter, G .; D. Ed., (1982
), John Wiley & Sons, Ltd. , 669-675, describe the development of an applicator device for this system that simultaneously mixes and applies the two components of the system. These combination systems and their uses have been extensively described: Seerich, T .; , J Head and Neck Pathol
(1982) 3: 65-69; O'Connor, A .; F. Et al. Otolary
ngo1 Head Neck Surg (1982) 90: 347-348;
See Marquet, J.M. , J Head and Neck Pathol (19
82) 3: 71-72; Thorson, G .; K. Et al., J Surg Onco
1 (1983) 24: 221-223. McCarthy, P .; M. May
o Clin Pros (1987) 62: 317-319 reports the addition of barium ions to this fibrin adhesive system in the treatment of bleeding duodenal cavity to facilitate follow-up monitoring. Portmann M. , J Hea
d and Neck Pathol (1982) 3:96;
See also, ibid, 94-95.

[0011] Efforts have also recently focused on ways to avoid health problems caused by the use of plasma-derived products in commercially available tissue adhesive products and systems. To this end, attempts have been made with varying degrees of success in isolating the corresponding portion of the fibrinogen-containing component. See, for example, Feldman, M .; C
. Et al., Arch Otolaryngol-Head and Neck Su
rg (1988) 114: 182-185; Feldman, M .; C. Et al.
ch Ophthalmol (1987) 105: 963-967; Feldm
an, M .; C. Et al., MJ Otolog (1988) 9: 302-305; S
ilberstein, L .; E. FIG. Et al., Transfusion (1988) 28
: 319-312. The use of autofibrinogen preparations is also obviously limited.

Collagen as a biomaterial. Collagen, the major connective tissue protein in animals, has a number of properties not found in synthetic polymers. Frequently quoted properties of collagen include good compatibility with living tissue, promotion of cell growth, and absorption and anabolism of implants (Shimizu, R. et al.
Biomat. Med. Dev. Art. Org. , 5 (1): 49-66 (1
977)). Various applications of this material have been tested: for example, dialysis membranes of artificial kidneys (Sterzel, KH et al., Ameri. Soc. Artif. Int.
. Organs 17: 293 (1971)), artificial cornea (Rubin, AL).
. Et al., Nature 230: 120 (1971) and U.S. Pat. No. 4,581.
030), the vitreous (Dunn, M. et al., Amer. Soc. Artif. I).
nt. Organs 17: 421 (1971)), artificial skin and blood vessels (Kr
ajicek, M.A. J. et al. Surg. Res. No. 4,290 (1964)) as a hemostatic agent (U.S. Pat. No. 4,215,200) and a soft contact lens (U.S. Pat. Nos. 4,264,155; 4,264,493; and 4). , 3
49,470; 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)).
.

However, natural collagen fibers are essentially insoluble in mature tissues due to covalent intermolecular crosslinks that convert collagen into an endless crosslinked network. Dispersion and solubility of native collagen can be achieved by treatment with various proteolytic enzymes, which break the intermolecular bonds and affect the basic, rigid, triple helical structure that gives the desired properties of collagen. Without removing the non-helical immunogenic terminal region (US Pat. Nos. 3,934,852; 3,121
No. 3,131,130; No. 3,314,861; No. 3,
530,037; 3,949,073; 4,233,360 and 4,488,911 also refer to general methods for preparing purified soluble collagen. ).

[0014] Various methods and materials have proposed collagens that have been modified to provide better biomedical adhesives (eg, De Toledo, AR, et al., Assoc. For Res. In Vision and). Ophthal
logic, Annual Meeting Abstract, Volume 31, Volume 3
17 (1990); Lloyd et al., "Covalent Bonding of Collagen and Acrylic Polymers", Amer.
ican Chemical Society Symposium on B
iomedical and Dental Applications of Polymers, Polymer Science and Techno
, Vol. 14, Plenum Press (Gebelin and Kob)
litz), New York, 1980, pp. 59-84; Shimizu 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) for a discussion of general collagen and synthetic polymers). In many cases, unmodified collagen-based adhesives have various disadvantages, including: (1) the heat-generating crosslinking / polymerization reaction of the exothermic reaction, (2) the long reaction. Time, and (3) reactions that cannot be performed in the presence of oxygen and in the physiological pH range (Lee ML et al., Adhesion in Biological Systems, RS
Manly, Academic Press, New York, 1970,
Chap. 17). Furthermore, many unmodified collagen-based adhesives contain toxic substances and are not suitable for biomedical use (eg, Buonoco
re, M .; G. FIG. (1970) and U.S. Patent No. 3,453,222).

[0015] In addition, collagen-based adhesives also have an immunological relationship, wherein the adhesives are derived from animal and typical bovine sources. Studies on the use of such collagens as injectable devices report a minor inflammatory response. More recently, the potential problem of transmitting disease to humans associated with bovine spongiform encephalopathy ("mad cow disease") has attracted attention, especially in Europe, where bovine source materials are limited.

Despite these deficiencies, certain collagen-based adhesives, which reportedly have adequate adhesive strength and utility in many medical applications, especially for soft tissues, have been described. ing. U.S. Pat. No. 5,219,895)
. These reports are consistent with the use of types I and II in collagen-based adhesives; where the purified type I and type II collagens are chemically modified to form monomers, which are Dissolve in the conditions and then polymerize to form a composition with adhesive and sealant properties. These reports are limited to collagen-based adhesives, which are composed of collagen from natural sources; and at the same time report mixtures of collagen. For example, type I proteins, as isolated from natural sources, typically contain about 10-20% of type III and other collagen and 90-80%, depending on the tissue source used. It is composed of type I collagen.

[0017] These reports do not mention type III collagen, the unexpected hemostatic properties of type III collagen or the use of recombinant collagen which can avoid the first chemical modification step.

[0018] 3. SUMMARY OF THE INVENTION A biologically compatible type III and / or type I collagen product with sealant and adhesive properties is a recombinantly derived soluble type III and / or
It can also be formed using a type I collagen monomer; where the monomer polymerizes to form a type III and / or type I collagen composition having adhesive and sealant properties. Preferably, the collagen has been derived using human and recombinant techniques. Type III collagen was selected for its unexpectedly superior hemostatic properties compared to other collagen types. Type I collagen was chosen for its structural properties. This polymerization reaction is carried out by using a suitable polymerization initiator (
For example, it can be started with a chemical oxidant, UV irradiation, a suitable oxidase or atmospheric oxygen).

In order to optimize the sealant and adhesive properties of the recombinant collagen product by optimizing the product's structural stability and hemostatic properties or its products,
The composition preferably comprises a combination of pure recombinant type I and type III collagen. The ratio of pure recombinant type III collagen to pure recombinant type I collagen is about 30% or less for type I collagen to about 30% or more for II.
It is type I collagen. More preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen is from about 70% to about 50% type I collagen to about 30% to about 50% type III collagen. . Most preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen is from about 70% to about 60% type I collagen to about 30% to about 40% type III collagen. .

It is an object of the present invention to provide a pure recombinant type III collagen tissue sealant, a pure type I tissue sealant or a pure recombinant type I and type III collagen tissue sealant. , They are free of other type I collagen and have the following properties and capabilities: (i) hemostasis. This sealant acts as a hemostatic barrier and reduces the risk of serum, lymph and fluid leakage. Where collagen type III has intrinsic hemostatic properties, its use in hemostatic devices provides an improvement over known fibrin sealants. Type I collagen also has certain hemostatic properties. (Ii) adhesion. Due to this adhesive property, the sealant atraumatically adheres by forming a strong bond therebetween, and conforms to uneven wound surfaces. This adhesive effect is increased by the combination of the reagents and collagen type III and / or type I as described below. (Iii) Wound healing. The sealant promotes fibroblast proliferation, which provides an improved healing process in a combination of efficient hemostasis and adhesion between the wound surfaces. Use of a composition according to the present invention, such as an anti-adhesive / wound healing composition, is expected to obtain normal (healing) tissue other than scar tissue (ie, optimal wound healing). Further, such compositions also reduce the inflammatory response.

Accordingly, an object of the present invention is to polymerize II as a safe and effective biological adhesive with appropriate adhesive strength for biomedical applications, especially for soft tissues.
It is to provide a type I and / or type I collagen composition. More specifically, the present invention relates to compositions useful for sealing punctures and incisions in organs, dermis and large blood vessels. Polymerized materials can assume a number of sizes and shapes consistent with their intended biomedical applications, including ophthalmology, plastic surgery,
Includes use in orthopedic and cardiac surgery.

In another object of the present invention, the type III and / or type I collagen composition further comprises a reagent for the sealant or the adhesive, which gives further desired properties. For example, fibrin, fibrinogen, thrombin, calcium ions, factor XIII can be included in the composition to better form a three-dimensional network of polymerized collagen. In yet another object of the present invention, the recombinant I
Type II collagen compositions incorporate drugs that have wound healing capabilities. In one embodiment, the drug is a connective tissue growth factor and is incorporated into a composition that slowly releases the drug to the wound.

[0023] 4. DETAILED DESCRIPTION OF THE INVENTION 4.1 (Definition) As used herein, the term "biologically compatible" refers to the term "biologically compatible collagen type III recombinant product" of the present invention. Refers to recombinant type III and / or I collagen that has been modified according to, has been incorporated into, implanted in, or located near a biological tissue of a subject, and More specifically, it does not appreciably degrade over time or induces an immune response or adverse tissue reactions following such integration or implantation or placement.

As used herein, the term “pure recombinant type I collagen” refers to human type I collagen produced by recombinant techniques substantially free of other types of collagen. I do. This term excludes type I collagen isolated from natural sources.

As used herein, the term “pure recombinant type III collagen” refers to human I produced by recombinant techniques substantially free of other types of collagen.
Reference is made to type collagen. This term excludes type III collagen isolated from natural sources.

4.2 Preparation of Polymerized Recombinant Type I and III Collagen Production of Type I and Type III Collagen Monomers Biologically Compatible Collagen Products with Adhesive and Hemostatic Properties of the Invention Useful types of collagen to form are collagen type I and type III collagen. Monomeric soluble type I and type III collagen is obtained by a recombinant process, including processes involving the production of type III collagen in transgenic animals. The above recombination process is described in US Pat. No. 5,593,859.
, Which is incorporated herein by reference. Preferably, type I or type III collagen uses at least one gene encoding a polypeptide comprising type I or type III collagen, and genes encoding the α and β subunits of the post-translation enzyme prolyl 4-hydroxylase. Thus, it is recombinantly produced by culturing the cells to be transfected and purifying the collagen monomers obtained therefrom. Preferably, the monomeric soluble type I and type III collagen material has consistency in consistency and clarity and clarity (
4 shows changes in transparency and clarity.

Polymerization of Type I and Type III Collagen Monomers The type I and type III recombinant collagen solutions can be subjected to substantially polymerization or cross-linking conditions to produce the polymerized collagen composition of the present invention. . The polymerization can be performed, for example, by using UV, gamma, or fluorescent irradiation. UV
Irradiation can be achieved in the short wavelength range using a standard 254 nm source or a UV laser source. Using a standard 254 nm source, 4-12 watts, the polymerization is 10-40
Minutes, preferably 20-30 minutes, exposure distance of 2.5-10 cm, preferably 2
. Occurs at a distance of 5-5 cm. Excessive UV exposure begins to depolymerize the collagen polymer. Polymerization using gamma irradiation can be performed using 0.5-2.5 Mrad. Excessive gamma exposure also depolymerizes the collagen polymer. Polymerization in the presence of oxygen can be performed by adding an initiator to the fluid prior to exposure.
Non-limiting examples of initiators include sodium persulfate, sodium thiosulfate, ferrous chloride tetrahydrate, sodium bisulfate, and oxidases (eg, peroxidase or catechol oxidase). If an initiator is used, the polymerization can take from 30 seconds to
It takes place within 5 minutes, usually 1 to 3 minutes.

[0028] The polymerizing agent is preferably UV radiation. However, while the polymerization or crosslinking of the monomer substituents can be performed by simply exposing the material to atmospheric oxygen, the rate of polymerization is much slower than with UV irradiation or chemical reagents.

[0029] Other agents may also be useful in the polymerization process. For example, to improve the cohesive strength of the adhesive formed from the composition of the present invention, a bifunctional monomeric crosslinker may be added to the monomer composition of the present invention that affects polymerization. Such crosslinkers are described in the art, for example, in US Pat. No. 3,940,362 (Ove).
rhults), which is incorporated herein by reference.

4.3 (Type I and Type III Collagen Compositions) The compositions of the present invention comprise polymerized type I and type III collagen, wherein:
The composition is produced by a process that includes the following steps: (1) production of type I and type III collagen monomers by the recombinant method described above; and (2)
Polymerization of such monomers.

For the purpose of optimizing the sealant and adhesive properties of the recombinant collagen product by optimizing the structural stability of the product and the hemostatic or product properties of the product, The product preferably comprises a combination of pure recombinant type I and type III collagen. The ratio of pure recombinant type III collagen to pure recombinant type I is about 30% or more of type III collagen to about 70% or less of type I collagen. More preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen is from about 30% to about 50% of type III collagen to about 7%.
0% to about 50% type I collagen. Most preferably, pure recombinant III
The ratio of type I collagen to pure recombinant type I collagen is from about 30% to about 40% II.
Type I collagen to about 70% to about 60% type I collagen.

[0032] The compositions of the present invention may be further comprised of other agents useful for adhering or sealing tissue. For example, in addition to the recombinant type I and / or type III collagen proteins, the composition preferably comprises factor XIII and / or fibrin / fibrinogen / fibronectin and / or plasminogen. Advantageously, the composition also includes a clotting enzyme (ie, thrombin,
In particular, in combination with divalent calcium (eg, calcium chloride). The concentration of calcium chloride can then vary, for example, between 40 mM and 0.2 M, depending on the particular purpose of the tissue adhesive composition, with high concentrations of calcium chloride inhibiting fibroblast proliferation. And therefore preferred in anti-adhesion applications (in the absence of fibronectin, which stimulates fibroblast proliferation). In addition, fibrinolysis inhibitors (
For example, plasmin inhibitors (eg, aprotinin, aprotinin, α
-2-antiplasmin, α-2-macroglobulin, α-1-antitrypsin, ε-
It may be of value to include aminocaproic acid or tranexamic acid)), or a plasmin activator inhibitor (eg, PAI-1 or PAI-2).

While the proportions of the previously known components in the tissue adhesive composition of the present invention can be selected with guidance of the prior art compositions, the required amount of viscosity enhancing polymer depends on the particular polymer and the intended It can be readily determined by one skilled in the art depending on the mode of use. Thus, if the concentration and / or molecular weight of the viscosity enhancing polymer is very low, the increase in viscosity is insufficient, and very high concentrations and / or molecular weight inhibit fibrin polymerization and adhesion to tissue.

[0034] By increasing the thrombin concentration, the polymerization of the composition of the present invention can be accelerated by a significant effect on time until the adhesive is deployed. For example, at low thrombin concentrations, the fibrin of the composition remains more or less liquid for several minutes after application. Thus, a further beneficial effect of increasing the viscosity with the viscosity-enhancing polymer according to the invention is that it may be used at low concentrations of thrombin, even if the part to be sealed is even on non-horizontal surfaces. Needed in situations that require later adaptation.

Similarly, a composition of the invention may be a fusion protein, rather than comprising a combination of the agents described herein, wherein type I and / or III
Type collagen as well as, for example, fibrin are combined to form one molecule. Such a fusion protein can be produced according to the recombinant techniques described herein.

In a further embodiment of the invention, the compositions of the invention are useful in wound healing by inducing or promoting the formation of tissue, or alternatively, by limiting the formation of fibrous adhesions. Including reagents. Such agents include antibiotics or growth factors (eg, connective tissue growth factor), which are described, for example, in US Pat. Nos. 5,408,040 and 5,585,270, The above references are cited herein by reference.

4.4 (Field of Use) Polymerized type III and / or type I collagen products can be useful for producing mechanical sealants and adhesive systems.

Tissue adhesive systems Areas of application are among others: ENT surgery, general surgery, dentistry, neurosurgery, plastic surgery, thoracic and vascular surgery, abdominal surgery, orthopedic surgery, disaster surgery, gynecology , Urology, and ophthalmology. The collagen sealants of the present invention have also been used for topical application of drugs such as antibiotics, growth factors and cell growth inhibitors.

(Sealing Film) In one aspect of the present invention, the polymerized collagen product may be formed in the form of a sealing film. Collagen-based films are consistently flexible and elastic, have the feel of a plastic film, and should show high biocompatibility. The use of a sealing film prevents the formation of adhesions after tendon surgery (ie, use as a wrap around the tendon), use as a synthetic eardrum, replacement of facial tissue tissue and replacement of wound dressing components. Including. Additional examples of potential uses for sealing films include:
Includes treatment of corneal detachment, wound closure, coating of catheters and instruments, and use as a substance to prevent adhesion formation in tissues other than tendons (eg, peritoneal cavity).

[0040] A further embodiment of the present invention includes a sealant and adhesive formulation, which comprises:
It can be used as a specific system for the delivery of numerous drugs and pharmaceutical compositions, including growth factors, antibiotics, and other biologically beneficial compounds. Such materials can be added to collagen adhesives or sealants to promote cell migration, cell adhesion, and wound healing.

Angioplasty and Angiography Angiography is a diagnostic procedure whereby a pigment is added to an artery, preferably a femur, to detect the presence or absence of coronary vascular disease. Injected into artery.
Angioplasty (also known as PCTA) is a therapeutic procedure, which involves inflation of a balloon in an artery (eg, a coronary artery) for the purpose of removing arterial obstruction. After puncturing the femoral artery, a balloon-catheter is introduced through the femoral artery and steered through the coronary arteries obstructed by atherosclerosis (plaque). Once deployed, the balloon is inflated and deflated several times in an attempt to open the artery by pushing fat against the vessel wall, allowing blood to circulate in the affected area of the myocardium. Various types of balloon catheters are commonly used for angioplasty and angiography, which are over-the-wire catheters.
ter) (Independent guide wire is placed at the site of disease (ride over
2) fixed wire catheter (combines a balloon catheter with a guidewire in one device); 3) rapid exchange or single operator exchange catheter (with a repetitive wire catheter that can be exchanged more conveniently than a standard repetitive wire catheter). Yes); 4) Includes a perfusion catheter (which allows blood flow during this procedure). The rotating tip catheter removes plaque formed on the artery wall. These devices utilize a technique called differential cutting. The calcified material is made into microscopic particles without damaging the artery due to the elastic properties of the artery wall.

Angioplasty is a more invasive and complex procedure than angiography. Since it requires the insertion of a sheath larger than that used in angiography. The sheath is used as a means to introduce the catheter into the artery. In addition, angioplasty also requires the use of a blood diluent (eg, heparin) to prevent clotting during and after the surgical procedure. Anticoagulants prevent the body's natural sealing / clotting mechanism, and thus sealing a puncture requires a significant amount of time.

According to the present invention, after removing the catheter and other invasive devices from the artery, an adhesive applicator can be inserted into the sheath, if necessary, and placed at a location near or in contact with the puncture in the artery. Is done. During the procedure, manual or mechanical pressure is applied to the artery to reduce blood flow at the puncture site. If possible, excess blood / fluid is removed from the puncture site. Subsequently, the type III and / or
Alternatively, the type I recombinant collagen monomer may be applied to the puncture on the external surface of the artery and / or within the puncture scar. The monomers may then be polymerized and / or cross-linked by the techniques described herein (eg, UV irradiation), thus polymerizing
It occurs within ~ 300 seconds, preferably within 0-120 seconds, more preferably within 0-30 seconds, and even more preferably within 3-10 seconds. By applying the collagen monomer composition to the outside of the artery, the incidence of embolism (obstruction of the artery or circulatory system) is substantially eliminated. Alternatively, polymerized type III and / or type I collagen can be used and the polymerization step can be avoided. Due to the bonding strength of the adhesive of the present invention, only a small amount of adhesive is needed to seal the punctured artery. Further, since the surgical adhesive according to the present invention can polymerize almost instantaneously, the adhesive can be applied on the surface of the artery and / or along the puncture of the artery without penetrating the interior of the artery. It can polymerize. Thus, large pieces or particles of material do 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 present invention, the patient need only be immobilized for a minimum amount of time.

4.5 (Administration) (Formulation) The tissue treatment composition of the present invention may be presented in a preparation of the same type as the prior art fibrin sealant. This component can be provided in ultra-low temperature frozen solution form or as a lyophilized powder, diluted with a suitable aqueous solution (eg, containing aprotinin and calcium ions, respectively) prior to use.

For compositions of the present invention that include a pharmaceutical reagent (eg, an antibiotic, growth factor, etc.), the collagen network formed upon application of the tissue adhesive by incorporating the reagent into the tissue adhesive Will be included. This ensures that the drug is retained at the site of application while being controllably released from the composition (eg, when used as eye drops, wound healing preparations, etc.). Also, as described above, the pharmaceutically active substance to be released from the tissue adhesive composition of the present invention may be a polymer having improved viscosity in itself or in a substance bound thereto.
Specific examples of such viscosity-enhancing polymers that meet the viscosity-enhancing requirements, and that may have therapeutic and pharmacological utility and that are desired to be biocompatible, include hyaluronic acid and Its salts and derivatives, which are readily soluble in water and, as noted above, have very short biological half-lives. Thus, the tissue treatment compositions of the present invention constitute an advantageous slow release preparation for proteoglycans (eg, hyaluronic acid and its salts and derivatives) and significantly increase their biocompatibility.

In particular, the compositions of the present invention are not limited in their adhesive properties, but also include non-adhesive compositions, especially if the composition is primarily intended for wound healing. The latter composition may specifically include non-adhesive proteins, such as albumin and / or growth factors. Substantially, a non-adhesive composition can also be obtained when the polymer portion of the composition interferes with the adhesive properties of the protein portion. It should be emphasized that the present invention includes both adhesive and substantially non-adhesive compositions herein, but often referred to herein as "adhesives" for reasons of simplicity. Called as.

(Application of Composition) The composition of the present invention can be applied using various dispensing devices. For example, surgical adhesive monitors the application process via an optical observation system,
U.S. Pat. Nos. 4,900,303 (Lemelson) and 5,372,303.
No. 585 (Tiesenbrunn).
The compositions of the present invention may also be applied by the device described in US Pat. No. 5,129,882 (Weldon et al.). The subject matter of these patents is incorporated herein by reference.

The composition according to the invention can also be applied in combination with other sealing means. For example, the adhesive may be applied to a puncture site (eg, visceral (eg, liver, gall bladder, intestine, stomach, kidney, heart, bladder, ureter, lung, esophagus, etc.) that is closed using surgical sutures or tapes. Puncture or incision). The adhesive in this case provides a complete seal, whereby fluid leakage from organs or blood vessels (for example,
The risk of leakage from the site of liver puncture). The surgical adhesive of the present invention may additionally be used in combination with other sealing means (eg, plugs, etc.). Such techniques are disclosed in U.S. Patent Nos. 4,852,568 (Kensey),
Nos. 612 (Kensey), 5,053,046 (Janese), 5,061,274 (Kensey), and 5,108,421 (Fowl)
er), No. 4,832,688 (Sagae et al.), No. 5,192,300
No. (Fowler), No. 5,222,974 (Kensey et al.), No. 5,27
No. 5,616 (Fowler et al.), No. 5,282,827 (kensey et al.), No. 5,292,332 (Lee), No. 5,324,306 (Mak)
No. 5,370,660 (Weinstein et al.) and 5,021,059 (Kensey et al.). The subject matter of these patents is incorporated herein by reference.

In particular, the compositions of the present invention can be used to bond two surfaces together by applying a particular composition to at least one of said surfaces. Depending on the specific requirements of the user, the adhesive composition of the present invention can be applied by known means (eg, a glass stir bar, sterile brush or infusion device), but in many situations, The pressurized aerosol distribution package preferably has the adhesive composition in solution with a compatible anhydrous propellant. Aerosol application of this monomer is particularly advantageous for applications in hemostasis.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C08H 1/00 C12P 21/02 C 4J011 C08L 89/00 C08F 2/46 4J040 C09J 189/00 2/48 C12N 15/09 A61K 37/12 C12P 21/02 37/02 // C08F 2/46 37/465 2/48 C12N 15/00 A (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU) TJ, TM), AL, AM, AU, AZ, BA, BB, BG, BR, BY, CA, CN, CU, CZ, EE, GE, GH, HR, HU, ID, IL, IS, JP, KG, KP, KR, KZ, LC, LK, LR, LT, LV, MD, MG, MK, MN, MX, NO, NZ, PL, RO, RU, SG, SI, SK, SL, TJ, TM , TR, TT, UA, UZ, VN, YUF term (reference) 4B024 AA01 BA08 BA80 CA02 DA02 GA11 HA03 4B064 AG01 CA10 CA19 CC24 DA01 4C084 AA02 AA03 BA44 CA53 DA40 DC10 DC20 DC50 MA02 MA67 NA14 ZA362 ZA892 4H10A30A CA40 EA34 FA72 FA74 4J002 AD001 GB01 GJ01 4J011 QA48 QB28 UA01 WA09 4J040 BA161 JA09 JB07 JB08 KA12 KA13 MA15 NA02 NA04

Claims (20)

[Claims] 1. A tissue adhesive or sealant composition containing polymerized type III collagen, wherein the adhesive or sealant composition recombinantly produces pure type III collagen monomer in cells. And a process of polymerizing the monomer with a reagent. 2. The composition according to claim 1, wherein said composition is biologically compatible. 3. The composition of claim 1, wherein the recombinant production of the type III collagen monomer comprises the following steps: (a) at least one gene encoding a polypeptide comprising type III collagen and prolyl. Culturing cells transfected with at least one gene encoding a polypeptide selected from the group of α or β subunits of 4-hydroxylase; and (b) purifying the type III collagen A composition comprising: 4. The composition of claim 1, wherein said monomer is polymerized using irradiation. 5. The composition according to claim 4, wherein said irradiation is UV irradiation. 6. The composition of claim 1, wherein said composition further comprises one or more reagents selected from the group of fibrin, fibrinogen, thrombin, factor XIII or connective tissue growth factor. 7. A process for producing a tissue sealant or adhesive, comprising:
A process comprising: (a) producing a type III collagen monomer by recombinant means; and (b) polymerizing the type III collagen monomer. 8. A tissue adhesive or sealant composition comprising polymerized type I collagen, wherein the adhesive or sealant composition recombinantly produces pure type I collagen monomer in cells. A composition produced by a process and polymerizing the monomer with a reagent. 9. The composition according to claim 8, wherein said composition is biologically compatible. 10. The composition of claim 8, wherein the recombinant production of the type I collagen monomer comprises the following steps: (a) at least one gene encoding a polypeptide comprising type I collagen and prolyl. Culturing cells transfected with at least one gene encoding a polypeptide selected from the group of the α or β subunits of 4-hydroxylase; and (b) purifying the type I collagen A composition comprising: 11. The composition according to claim 8, wherein the monomer is polymerized using irradiation. 12. The composition according to claim 11, wherein said irradiation is UV irradiation. 13. The composition of claim 8, wherein said composition further comprises one or more reagents selected from the group of fibrin, fibrinogen, thrombin, factor XIII or connective tissue growth factor. 14. A process for producing a tissue sealant or adhesive, comprising the following steps: (a) producing type I collagen monomer by recombinant means; and (b) said type I collagen monomer A process comprising the step of polymerizing. 15. A tissue adhesive or sealant composition comprising polymerized pure type III collagen and polymerized pure type I collagen. 16. The composition according to claim 15, wherein said composition is biologically compatible. 17. Pure recombinant III against pure recombinant type I collagen.
The ratio of type I collagen is about 70% or less and about 30% or more I type collagen.
16. The composition of claim 15, which is a type II collagen. 18. The composition according to claim 15, wherein said monomer is polymerized using irradiation. 19. The composition according to claim 15, wherein said irradiation is UV irradiation. 20. The composition according to claim 15, wherein said composition further comprises one or more reagents selected from the group of fibrin, fibrinogen, thrombin, factor XIII or connective tissue growth factor.