Collagen peptide-based pharmaceutical composition and use thereof

Abstract

The present invention is in the field of pharmaceutical chemistry, biotechnology and pharmacy. The present invention provides compositions comprising one or more collagen-mimetic peptides, optionally linked to one or more therapeutic compounds or one or more imaging compounds; methods for using such compositions for the treatment, prevention, amelioration, cure and / or diagnosis of certain diseases and physical disorders of humans and veterinary animals, in particular diseases, disorders and injuries of the nervous system and of the cardiovascular system. The present invention also provides the use of such compositions in the treatment, prevention, amelioration, cure and / or diagnosis of a variety of other diseases, disorders and conditions in tissues, organs and organ systems. The present invention also provides medical devices comprising one or more such compositions of the present invention.

Description

Cross-references to related applications and their inclusion by reference

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 594,245, filed October 30, 2023, entitled “Pharmaceutical Compositions Based on Collagen Peptides and Use Thereof,” and whose inventors are Richard E. Schlumpf, Brian J. Del Buono, Robert O. Baratta, and David J. Calkins, the entire contents of which are incorporated herein by reference. Statement Regarding Federal Government-Sponsored Research and Development

[0002] not applicable. Names of the parties to the joint research agreement

[0003] not applicable. References to sequence lists submitted electronically

[0004] not applicable. Technical Field

[0005] This invention pertains to the fields of medicinal chemistry, biotechnology, pharmaceuticals and medical devices, as well as the use of pharmaceutical compounds and medical devices for the treatment, prevention and improvement of diseases, conditions and physical discomforts in humans and animals, particularly certain neurological diseases and conditions, including those involving the central and peripheral nervous systems, and certain cardiovascular diseases and conditions. Background Technology

[0006] Collagen is the most abundant protein in vertebrates and is the basic structural protein of vertebrate tissues. It is found in almost all tissues, including skin and other epithelial tissues (including the lining of most intracavitary organs such as the gastrointestinal tract), tendons, bones, blood vessels, cartilage, ligaments, and teeth. In humans, collagen accounts for about one-third of total protein and about three-quarters of the dry weight of skin (see Shoulders, MD and Raines, RT, Ann. Rev. Biochem. 78:929-958 (2009); Gelse, K., et al., Adv. Drug Deliv. Rev. 55:1531-1546 (2003)).

[0007] Collagen is a fibrous protein composed of triple helices, typically consisting of two identical chains and a third chain with a slightly different chemical composition. Mammals produce at least 46 different collagen polypeptide chains, which combine to form variants or "types" of collagen. To date, 28 types of collagen have been described. Collagen types are generally grouped according to their structural form: fibrous (types I, II, III, V, and XI) account for approximately 90% of all collagen found in mammals; non-fibrous (basement membrane or type IV, and other non-fibrous collagen types with interrupted helical structures, see above). The five most common collagen types and their tissue distribution are as follows:

[0008] Type I: Skin, tendons, organs, bones, blood vessels, and connective tissue;

[0009] Type II: Cartilage;

[0010] Type III: reticular connective tissue, often associated with type I collagen;

[0011] Type IV: Basement membrane of epithelial tissue and certain solid tumors; and

[0012] V-shape: hair, placenta, outer cell membrane.

[0013] In each of these variants, the collagen polypeptide chain consists of approximately 300 amino acid repeats of proline (Pro), 4(R)-hydroxyproline (Hyp), and glycine (Gly), typically in an XY-Gly sequence, where X is usually a Pro residue and Y is usually a (Hyp) residue; in vertebrates, the typical repeating motif in collagen is ProProGly (see Hulmes, DJS, “Collagen Diversity, Synthesis and Assembly,” in: Collagen: Structure and Mechanics, P. Fratzl, ed., New York: Springer, pp. 15–47 (2008)). Subsequently, in vivo, the hydroxylation of the Pro residues occurs enzymatically after collagen biosynthesis but before the chain begins to form a triple helix. Therefore, hydroxylation of at least one Pro residue in the ProProGly motif (typically forming ProHypGly) appears to be important for both the proper folding and stability of the collagen triple helix, both of which are essential for the normal structure and function of collagen in vivo (see Shoulders, MD and Raines, RT, Ann. Rev. Biochem. 78:929-958 (2009)). For example, (ProHypGly) 10The unwinding temperature of the triple helix of (SEQ ID NO: 396) chain is 58°C, while (ProProGly) 10 The unwinding temperature of the triple helix of the (SEQ ID NO: 397) chain is only 24°C (Sakakibara et al., Biochim. Biophys. Acta, 303:198-202 (1973)), and (ProHypGly) 10 The rate at which the (SEQ ID NO: 396) chain folds into a triple helix is ​​significantly greater than that of (ProProGly). 10 The corresponding rate of the chain (SEQ ID NO: 397) (Chopra and Ananthanarayanan, Proc. Natl. Acad. Sci. USA, 79:7180-7184 (1982)).

[0014] Type I collagen is the most abundant and well-studied type of collagen. In humans and most other animals, it constitutes more than 90% of the organic mass of bones and is the main collagen in tendons, skin, ligaments, cornea, and many interstitial connective tissues (with very few exceptions such as hyaline cartilage, brain, and vitreous humor). The triple helix of type I collagen is usually formed by two identical α1 chains and one α2 chain forming a heterotrimer. The triple helix fibers are mainly incorporated in the body into complex fibrils containing other types of collagen, as mentioned above, which vary according to tissue type and location (Fleischmajer, ED et al., J. Struct. Biol. 105: 162–169 (1990); Niyibizi, C. and Eyre, DR. Connect. Tissue Res. 20: 247–250 (1989)). In most organs, especially in tendons and fascia, type I collagen provides tensile stiffness, while in bones, it determines biomechanical properties related to load-bearing capacity, tensile strength, and torsional stiffness.

[0015] In connective tissues such as bone, tendons, cartilage, ligaments, skin, blood vessels, and teeth, individual collagen molecules are tightly wound together in triple helices. These helices are organized into fibrils with high tensile strength through the cross-linking of individual triple helical fibers (Lodish, H. et al., “Collagen: The Fibrous Proteins of the Matrix”, Molecular Cell Biology, 4th ed., Section 22.3, New York: WH Freeman (2000)). Altering the arrangement and cross-linking of collagen fibrils enables vertebrates to withstand one-dimensional (tendons), two-dimensional (skin), or three-dimensional (cartilage) stresses.

[0016] Collagen plays a crucial role in maintaining the structural integrity of tissues and organs in the body. In all solid organs, collagen is a major component of the interstitial matrix and basement membrane, while in all connective tissues, particularly bone and cartilage, it forms the main functional framework. However, beyond biomechanical functions, collagen participates in a variety of other functions. For example, specific cell surface and intracellular receptors interact with collagen, and the signaling of these receptors is involved in cell adhesion, differentiation, growth, and other cellular activities, as well as cell survival in vivo and in vitro (Vogel, WF, Eur. J. Dermatol. 11: 506-514 (2001); Gelse, K., et al., Adv. Drug Deliv. Rev. 55:1531-1546 (2003)). Collagen also participates in the capture, local storage, and delivery of growth factors and cytokines in various tissues where collagen is present. Through these receptor interactions and storage and delivery functions, collagen plays a crucial role in organ development, wound healing, and tissue repair (Chattopadhyay, S. and R. Raines, Biopolymers 101: 821-833 (2014); Yamaguchi, Y. et al., Nature 346: 281-284 (1990); Hay, ED, J. Cell Biol. 91:205s-223s (1981); Bautista, CM et al., Metabolism 39: 96-100 (1990); Zhu, Y. et al., J. Cell Biol. 144: 1069-1080 (1998); Schlegel, KA Chattopadhyay, S. et al., J. Tissue Eng. Regen., Biomaterials 25:5387-5393 (2004); Kumar, VA, et al., Biomacromol. 15: 1484-1490 (2014)).These functions also qualify collagen as a candidate transport medium for delivering therapeutic compounds (see, for example, Chattopadhyay, S. et al., J. Tissue Eng. Regen. Med. 10:1012-1020 (2012); Schuppan, D. et al., Gastroenterol. 114: 139-152 (1998); Frenkel, SR et al., J. Bone Jt. Surg. 79-B: 831-836 (1997); Albu, MG et al., “Collagen-Based Drug Delivery Systems for Tissue Engineering”, in: Biomaterials Applications for Nanomedicine, Pignatello, R. (ed.), ISBN: 978-953-307-661-4, DOI: 10.5772 / 22981, Rijeka, Croatia: InTech, URL: [https: / / www.intechopen.com / books / biomaterials-applications-for-nanomedicine / collagen-based-drug-delivery-systems-for-tissue-engineering (2011)](https: / / www.intechopen.com / books / biomaterials-applications-for-nanomedicine / collagen-based-drug-delivery-systems-for-tissue-engineering (2011)), and for use in wound healing by directly promoting tissue repair or regeneration (Wakitani, S. et al., J. Bone Jt. Surg. 71-B: 74–80 (1989); Kumar, VA et al., Biomacromol. 15: 1484-1490 (2014)). Collagen (more specifically, damaged collagen) is also associated with tumor progression and metastasis in humans and other vertebrates (for a review on this issue, see Fang, M. et al., Tumor Biol. 35:2871-2882 (2014)).

[0017] However, in addition to intact collagen molecules, collagen fragments may also have potential therapeutic uses and may actually function in a better way than natural collagen. For example, non-collagenous fragments of type IV, XV, and XVIII collagen have been shown to promote the growth of angiocytes and tumor cells and affect a variety of other cellular activities (Ortega, N. and Werb, Z., J. Cell Sci. 115: 4201-4214 (2002); Davis, GE et al., Am. J. Pathol. 156: 1489-1498 (2000); O'Reilly, MS et al., Cell 88: 277-285 (1997)). Similarly, as described in more detail below, the efficacy of fragments of type I collagen or synthetic collagen mimic peptides (CMPs) in treating diseases and medical conditions has been investigated, both as active pharmaceutical ingredients (APIs) and for delivering skin wound healing agents (see U.S. Patent Nos. 5,973,112, 7,122,521, 7,858,741 and U.S. Patent Publication No. US 2007 / 0275897 A1, the disclosures of all these patents being incorporated herein by reference in their entirety; see also, for example, Chattopadhyay, S. et al., J. Tissue Eng. Regen. Med. 10:1012-1020 (2012); Kumar, VA et al., Biomacromolecules 15:1484-1490 (2014)).

[0018] Collagen abnormalities are associated with a variety of human diseases, including eye diseases and conditions such as cataracts and glaucoma (Coudrillier, B., et al., PLoS ONE 10: e0131396 (2015); Huang, W., et al., Med. Sci. Monit. Basic Res. 19: 237-240 (2013); Dua, HS, et al., Br. J. Ophthalmol. 98: 691–697 (2014)), arthritis, rheumatism, osteoporosis, atherosclerosis, and cirrhosis. Collagen destruction is also associated with certain human and animal diseases, such as certain cancers (particularly endoscopic organ cancers and certain sarcomas); see, for example, Lauer, JL, and Fields, GB, “Collagen in Cancer”, published in [Journal Name - missing from original text]. The Tumor Microenvironment, New York: Springer, pp. 477-507 (2010). Collagen is also essential for wound healing and is known to be upregulated in areas of healing epithelial wounds (see, for example, U.S. Patent Nos. 5,973,112 and 7,122,521, which are incorporated herein by reference in their entirety; see also Chattopadhyay, S., et al., J. Tissue Eng. Regen. Med. 10:1012-1020 (2012); Chattopadhyay, S., et al., Org. Biomol. Chem. 10:5892-5897 (2012); Kumar, VA, et al., Biomacromol. 15:1484-1490 (2014)), including in the skin and cornea. In fact, certain peptides of collagen, collagen fragments, or natural collagen have been reported to show promising potential in treating certain wounds and diseases in humans and animals, particularly skin wounds (see, for example, U.S. Patent Nos. 5,973,112, 7,122,521, 7,858,741 and U.S. Patent Publication No. US 2007 / 0275897 A1, the entire contents of which are incorporated herein by reference; see also Kumar, VA et al., Biomacromolecules 15:1484-1490 (2014)). These collagen fragments or collagen mimic peptides are believed to specifically target areas of collagen damage associated with skin wounds by inserting into damaged collagen and reforming the natural collagen I triple helix (see, for example, Chattopadhyay, S., et al., J. Tissue Eng. Regen. Med. 10:1012-1020 (2012); Chattopadhyay, S., et al., Org. Biomol. Chem. 10:5892-5897 (2012)). Therefore, collagen has been used as a delivery medium for certain drugs with varying degrees of success (see, for example, B. An, et al., Adv. Drug Deliv. Rev. 97:69-84 (2016); V. Chak, et al., Intl. J. Pharm. Teaching and Practices 4:811 (2013)).Collagen mimic peptides have also been used for topical application to deliver bound therapeutic compounds (neuropeptides called substance P) to skin wound areas; this CMP-substance P conjugate has been shown to accelerate wound healing in mouse skin models (Chattopadhyay, S., et al., J. Tissue Eng. Regen. Med. 10:1012-1020 (2012)). Certain extracellular matrix (ECM) components, including collagen, are also involved in maintaining the normal structure and function of the nervous system, particularly the peripheral nervous system, and disruption or damage to these ECM components often leads to neuronal disorder and / or death (see, for example, Koopmans G, Hasse B, Sinis N. The role of collagen in peripheral nerve repair (Chapter 19). International Review of Neurobiology. Vol. 87: Academic Press, Elsevier; pp. 363–79 (2009); Gao X, et al., Rev. Neurosci. 24(4):443-53 (2013); Campbell IC et al., J. Biomech. Eng. 136(2):021005 (2014); Vecino E et al., J. Cytol. Histol. S3:007 (2015); Vecino E., and Kwok, JCF, “The Extracellular Matrix in the NervousSystem: The Good and the Bad”). "Aspects", in Composition and Function of the Extracellular Matrix in the Human Body, F. Travascio, ed., Intech Open, ISBN 978-953-51-2416-0 (2016), accessed on November 8, 2019 at http: / / dx.doi.org / 10.5772 / 62527.

[0019] Treatments for diseases / conditions are expensive, difficult to deliver specifically, and may have harmful effects at sites far from the intended site of action. For example, many pharmaceutical compositions, including antibiotics, small molecule therapeutics (e.g., anticancer compounds), and biologics (e.g., monoclonal antibody therapeutics), are administered parenterally in a non-targeted manner; they must diffuse or otherwise locate the site of pain before providing therapeutic benefit. This “shotgun” approach inevitably requires higher dosing rates and may result in longer treatment durations and lower patient compliance compared to treatments that would deliver therapeutic compounds and compositions in a more targeted manner, allowing for controlled or programmable release at or near the site of pain in humans or animals. In particular, eye diseases, conditions, and physical ailments often prove difficult to treat and / or remedy. For example, presbyopia is an eye condition that is often associated with aging (Lafosse, E. et al., Cont. Lens Ant. Eye 43(2): 103-114 (2020); Balgos, MJTD et al., Taiwan J. Ophthalmol 8:121-140 (2018); Katz, JA et al., Clin. Ophthalmol. 15:2167-2178 (2021)), and is usually treated with optical correction, including mechanical correction (e.g., by eyeglasses or contact lenses) or refractive surgery (e.g., conductive keratoplasty, LASIK / LASEK surgery, photorefractive keratotomy, or intraocular lens implantation).Myopia is a common eye condition affecting all ages, including children (Hou, W. et al., Eye Contact Lens 44(4):248-259 (2018); Lau, JK et al., Invest. Ophthalmol. 61(2):22 (2020); Tideman, JWL et al., Acta Ophthalmol. 96:301-309 (2018)) and adults (Kim, HK et al., Int. J. Ophthalmol. 14(8):1231-1236 (2021); Wang, B. et al., PLoS ONE 12(4):e0175913 (2017); Pugazhendhi, S. et al., Clin. Ophthalmol. 14:853-873). (2020)), which is treated with corrective lenses and / or medications, such as atropine or pirenzepine (Gwiazda, J., Optom. Vis. Sci. 86(6):624-628 (2009)) and other drug treatments (Wang, W.-Y. et al., Biomed. Pharmacother. 133:111092 (2021)). Especially for children and the elderly, due to inconvenience, discomfort and other reasons, patients often cannot fully comply with these corrective measures, resulting in continued vision loss, and in some cases even worsening.

[0020] Previous work by some of the inventors has demonstrated and / or suggested that certain CMP-containing formulations may be used to treat certain anterior eye diseases, such as dry eye and other corneal diseases, conditions and injuries (see, for example, U.S. Patent No. 10,632,168; Baratta, RO et al., Front. Pharmacol. 12:705623 (2021); Baratta, RO et al., Surv. Ophthalmol. 67:60-67 (2022)) and certain posterior eye diseases and conditions, such as glaucoma, macular degeneration, optic neuropathy, etc. (see, for example, U.S. Patent Publication No. US 2020 / 0353056 A1; McGrady, NR et al., Front. Pharmacol. 12:764709 (2021); Ribeiro, M. et al., Int. J. Mol. Sci 23: 2911 (2022); Ribeiro et al., Int. J. Mol. Sci. 23: 7004 (2022)); the entire contents of each of these references are incorporated herein by reference. However, this prior work has largely failed to address the potential impact of CMP-containing formulations on the treatment and / or prevention of other ocular conditions such as myopia, presbyopia, and keratoconus, most of which require mechanical and / or surgical interventions that are often incomplete, uncomfortable, or inconvenient for patients and often require adjustment and / or re-intervention as patients age.

[0021] Age-related neurodegenerative diseases associated with acute damage to the central nervous system (CNS) are an increasingly debilitating burden on those struggling with them, caregivers of such patients, and healthcare systems that must provide long-term care. Most pharmacological or gene therapy approaches that protect or repair neurons and their circuits in the CNS modulate the action of one or more receptor-ligand or intracellular signaling pathways associated with pathogenic pathways (to slow progression) or nutritional support (to combat disease progression). However, the extracellular space may offer equally abundant but underdeveloped opportunities for novel therapeutic approaches. The extracellular space is a vital component of all tissues and organs, including the brain. A more accurate description might be extracellular material, as this space is filled, among other components, with a complex and highly dynamic network of structural and bioactive proteins that make up the ECM, particularly collagen, which is primarily produced by astrocytes, neurons, and vascular cells in the CNS. Historically, collagen has been primarily viewed as a scaffold protein that increases biomechanical stability. However, the ECM is also involved in maintaining cell and tissue homeostasis and regulating a variety of cellular activities that play a key role in maintaining the health and normal function of cells, tissues, organs, and organ systems.

[0022] Therefore, there is a need in the art for formulations and methods of administration that overcome many of the drawbacks of conventional treatments for certain eye diseases and conditions in humans and animals. Such formulations and methods of administration would allow for the use of lower doses of the drug and more targeted delivery of the drug to the intended site of action, as well as reduce treatment problems or delays due to patient non-compliance. Finally, there is a need in the art for methods of producing such compositions that would meet the needs of medical and patient populations to maximize therapeutic efficacy while reducing costs. Summary of the Invention

[0023] The inventors deduced that, since collagen destruction is associated with a variety of diseases and conditions in humans and other animals, conjugating various therapeutic and / or diagnostic compounds with collagen or collagen-mimicking peptides would provide an elegant, rapid, and reproducible method to overcome many of the aforementioned limitations in the treatment and diagnosis of certain physical diseases and conditions, as well as in drug delivery. Therefore, the present invention provides compositions suitable for methods of treating and diagnosing physical conditions and diseases, and provides drug delivery systems, medical devices, and methods of manufacturing thereof. Thus, the present invention fulfills the aforementioned needs in the art.

[0024] In one aspect, the present invention provides compositions comprising one or more collagen mimic peptides (CMPs), wherein in certain embodiments, the collagen mimic peptides have been conjugated with one or more therapeutic compounds and / or one or more diagnostic compounds to form CMP conjugates and compositions. Such CMPs and CMP conjugates, as well as compositions comprising such CMPs and / or CMP conjugates, can be used to treat, prevent, improve, and diagnose various diseases, conditions, and physical conditions in humans and animals. In certain embodiments of this aspect, the present invention provides compositions comprising such CMPs and / or CMP conjugates and one or more pharmaceutically acceptable carriers, excipients, or complexes, and optionally one or more additional therapeutic or diagnostic agents, to provide therapeutic and diagnostic compositions that can be used to treat, prevent, improve, or diagnose certain diseases and conditions in humans and animals.

[0025] In another aspect, the present invention provides a method for treating, preventing, improving, or diagnosing certain diseases and conditions in humans and animals by applying the conjugates and / or compositions of the present invention to humans or animals suffering from or susceptible to such diseases or conditions. Diseases and conditions suitable for treatment, prevention, cure, improvement, or diagnosis according to this aspect of the invention include eye diseases or conditions, skin diseases or conditions, cancer, gastrointestinal diseases or conditions, genitourinary tract diseases or conditions, fibrotic diseases or conditions, cardiovascular diseases or conditions, bone diseases or conditions, and rheumatic diseases or conditions.

[0026] In another aspect, the present invention provides a medical device coated with or comprising one or more conjugates or compositions of the present invention. In a related aspect, the present invention provides a method for treating, curing, preventing, or improving a disease or symptom in a human or animal, comprising implanting one or more medical devices of this aspect of the present invention into a human or animal under conditions of treating, curing, preventing, or improving the disease or symptom.

[0027] In other respects, the present invention provides methods for manufacturing the compositions, conjugates, and medical devices of the present invention.

[0028] Other objects, advantages and features of the invention will become apparent to those skilled in the art upon reading the description, drawings, examples and claims set forth herein. Attached Figure Description

[0029] Figure 1 These are a series of inverted bright-field micrographs of dorsal root ganglion (DRG) neurons plated onto tissue culture plates that have been treated with type I collagen (…). Figure 1 A, 1E) or type I collagen digested with MMP-1 ( Figure 1 B-1D and 1F-1H) were coated overnight. After coating, the plate was coated with a medium (PBS; Figure 1 B, 1F) or CMP 10A (SEQ ID NO:473) Figure 1 C, 1G) or CMP 9C (SEQ ID NO:5) Figure 1 The DRG explant culture was treated with D and 1H, and then plated with DRG explants. The DRG explant culture was incubated at 37°C, and then plated for 24 hours (t=24 hours later). Figure 1 A-1D) and t=48 hours ( Figure 1 Photographs were taken using E-1H.

[0030] Figure 2 This is from the text above. Figure 1 A pair of high-magnification micrographs of DRG explants were used to measure explant growth. Figure 2 A: Real-time visualization of DRG explants 48 hours after plating on intact collagen using multifocal high-magnification phase-contrast microscopy to optimize the identification and measurement of the longest neurite (arrow). Figure 2 B: Confocal micrograph of an explant-like structure showing a neurite (red) extending from a NeuN-labeled DRG neuron (green) with an immunolabeled β3-tubulin layer. Scale bar: 250 µm Figure 2 A) or 100 µm ( Figure 2 B).

[0031] Figure 3These are a pair of bar graphs showing neurite growth in DRG neuron explants after CMP treatment. These graphs show the longest neurite growth at 48 hours. Figure 3 A) and the area of ​​the explant growth region ( Figure 3 B), whose values ​​relative to intact collagen were normalized (n=178). Compared with intact collagen (#), explants plated on digested collagen and treated with only the medium (n=139) showed significantly reduced neurite length (p=0.014) and region area (p<0.001).

[0032] Figure 4 It is an intravitreal injection of a Tide Fluor™2-conjugated (Pro-Pro-Gly)7 (SEQ ID NO:1) CMP (“TF2-CMP”) solution. Figure 4 A, 4B) or ocular surface application ( Figure 4 A series of confocal fluorescence micrographs of mouse eyes (C, 4D). Mice were sacrificed on day 3 after topical application of TF2-CMP or on the day of intravitreal injection to determine the localization of TF2-CMP. Figure 4 A and 4C show the extranuclear localization of TF2-CMP in the ganglion cell layer of the retina (arrow: retinal vessels; triangular head: ganglion cell nucleus), while Figure 4 B and 4D show the location of TF2-CMP in or near the internal limiting membrane (vitreous surface) of the retina.

[0033] Figure 5 The results of CMP treatment in mice on neural structures in the corneal nerve bed are shown. Figure 5 Image A depicts representative pseudo-color images of corneal nerve beds from untreated, mediated, and CMP 03A (SEQ ID NO:1) mice. These images show the extent of collagen fragmentation in the subbasal plexus (left-handed column) and epithelial terminals (central column). Dashed boxes in the micrographs of the central column indicate the location of magnified insert images (right-handed column). Figure 5 B is a bar graph showing the level of nerve growth in a micrograph taken from the central subbasal layer contrasting with the epithelial layer.

[0034] Figure 6 These are a series of fluorescence micrographs showing that CMP promotes axonal repair after nerve compression. Figure 6A: A stitched-together montage of confocal micrographs of longitudinal sections of the optic nerve two weeks after compression, from an eye that received a mediator (DMOS) intravitreal injection three days after injury. Axons containing cholera toxin B (CTB, false-color white) extend to the site of compression (dashed line) but generally do not extend distally toward the brain (arrow). Astrocytes labeled with glial fibrillary acidic protein (GFAP, red) are shown for comparison. Figure 6 B: In contrast, CTB-containing axons in nerve slices from the eye that received CMP 13A (SEQ ID NO:6) extended beyond the compression site and were even visible at more distal locations along the nerve. The repaired axons largely overlapped with the localized plaques of CMP 13A (green), visualized by the fluorophores to which they were attached. Scale bar = 200 μm ( Figure 6 A) or 100 μm ( Figure 6 B).

[0035] Figure 7 These are a series of bar graphs showing the effect of CMP treatment on axonal recovery after nerve compression. These graphs show the number of CTB+ axonal segments at a specific distance distal to the injury site two weeks after nerve compression in each sample. Figure 7 The symbols in A and 7B), which are numerically normalized relative to nerve width. At each location, the median score of axons (dashed lines) in the nerves of the eye treated with CMP 13A (SEQ ID NO:6) is given. Figure 7 A) Exceeding the median value in nerves from the eye treated with DMSO (DMSO) Figure 7 B); This trend is summarized in Figure 7 In the line drawing illustration in B. Figure 7 C describes something higher than that from Figure 7 The median (first 50%) number of CTB+ axon segments in A and B was averaged at the distance from the indicated compression site. Figure 7 D and 7E depict the mean length (7D) and the length of the longest segment (7E) of axonal segments from eyes treated with CMP or mediators, indicating that axonal segments from nerves from eyes treated with CMP extend further than those from eyes treated with mediators, as reflected in the mean length and the 25 longest segments in each group (*, p<0.001).

[0036] Figure 8 It is derived from wild-type (“WT”) mice or from ApoE mice. - / -A series of fluorescence micrographs of mouse liver sections were used to examine the localization of mediators (PBS) or fluorescently labeled CMPs administered via tail vein injection. Sections were stained with DAPI (for nuclear visualization) and examined by fluorescence microscopy at emission wavelengths of 405 nm (for DAPI) or 488 nm (for fluoro-CMP). Row A: DAPI visualization only; Row B: DAPI+CMP visualization; Row C: CMP visualization only; Row D: A higher magnification view from the inset (white box) in row C. Column 1: WT mice treated with mediators only; Column 2: WT mice treated with 100 µM CMP; Row 3: ApoE mice treated with 100 µM CMP. - / - Mice.

[0037] Figure 9 yes Figure 8 The images depicted in rows C and D, and column 3, are higher magnification fluorescence micrographs. The arrows indicate fluorine-labeled CMP in ApoE. - / - Accumulation in the basement membrane of blood vessels in liver slices from mice.

[0038] Figure 10 It comes from wild-type mice ( Figure 10 A) or from ApoE - / - mice ( Figure 10 B) A pair of fluorescence micrographs of aortic arch sections, used to examine the localization of fluorescently labeled CMPs after injection via the tail vein.

[0039] Figure 11 It comes from the wild type ( Figure 11 A) or ApoE - / - mice ( Figure 11 B) High-power view of the aortic arch, such as Figure 10 The process involved examining CMP localization, which also revealed occlusion of the aortic arch co-located with the CMP (arrow), indicating that ApoE mice, compared to wild-type mice, exhibited... - / - Collagen in mice is destroyed.

[0040] Figure 12 It comes from wild-type mice ( Figure 12 A, 12B) or ApoE treated with the antibiotic streptozotocin (STZ) - / - A series of fluorescence micrographs of mouse retinal sections, showing the presence of the antibiotic streptozotocin, a compound exhibiting preferential toxicity to pancreatic β-cells and inducing diabetes and accelerating the progression of atherosclerosis. The localization of fluorescently labeled CMPs (green) in the sections after tail vein injection, the treatment of labeled reactive glial astrocytes with GFAP, and the treatment of labeled blood vessels with isolectins.

[0041] Figure 13 It comes from wild-type mice ( Figure 13 A-13C) or as Figure 12 ApoE- / - mice treated with STZ (shown) Figure 13 A series of fluorescence micrographs of mouse brain sections (D-13F). The localization of fluorescently labeled CMPs (green) after tail vein injection was examined in the sections, as well as reactive glial astrocytes labeled with GFAP and blood vessels labeled with fluorescent biotin. Analysis was performed on sections from the cerebral cortex (…). Figure 13 A, 13D), hippocampal CA1 area ( Figure 13 B, 13E) and dentate gyrus ( Figure 13 Ligand localization in two mouse phenotypes using C and 13F sections. Scale bar = 100 μM

[0042] Figure 14 It comes from treatment with STZ followed by PBS mediator via tail vein injection. Figure 14 A-14C) or CMP ( Figure 14 A series of fluorescence micrographs of mouse brain sections from ApoE- / - mice treated with D-14F. PECAM-1 (CD31) expression in the sections was examined by immunolabeling, an indicator of intact blood vessels. Origin from the cerebral cortex was determined by fluorescence confocal microscopy. Figure 14 A, 14D), hippocampal CA1 area ( Figure 14 B, 14E) and dentate gyrus ( Figure 14 Anti-CD31 labeling of thin sections of C, 14F. Scale bar = 100 μM.

[0043] Figure 15 This shows the total CD31 staining area in the cerebral cortex, hippocampal CA1, and dentate gyrus (DG) regions of the brains of ApoE- / - mice treated with STZ and subsequently treated via tail vein injection of PBS mediator or CMP. Figure 15 A) and staining intensity ( Figure 15 B) A series of bar charts. Data are expressed as mean ± SEM. *p≤0.02; **p=0.001; ****p<0.0001; ns=not significant (p=0.09); two-way ANOVA.

[0044] Figure 16 It comes from treatment with STZ followed by PBS mediator via tail vein injection. Figure 16 A-16C) or CMP ( Figure 16A series of fluorescence micrographs of brain slices from ApoE- / - mice treated with D-16F and then treated with sulfonyl-NHS biotin one day before euthanasia to track vascular leakage. Biotin was localized in the thin slices using streptavidin Alexa-647, and the biotin immunolabeling of the slices was examined by fluorescence confocal microscopy. Scale bar = 100 μM.

[0045] Figure 17 This shows the biotin staining intensity in the cerebral cortex, hippocampal CA1, and dentate gyrus (DG) regions of the brains of ApoE- / - mice treated with STZ and subsequently treated via tail vein injection of PBS mediator or CMP. Figure 17 A) and CD31 staining intensity normalized relative to biotin intensity ( Figure 17 B) is a series of bar charts. Data are represented as mean ± SEM. Figure 17 A: *p≤0.02; **p=0.01; Two-way ANOVA. Figure 17 B: *p=0.04; Two-way ANOVA.

[0046] Figure 18 It comes from treatment with STZ followed by PBS mediator via tail vein injection. Figure 18 A-18C) or CMP ( Figure 18 A series of fluorescence micrographs of mouse brain sections from ApoE- / - mice treated with D-18F. Type IV collagen expression in the sections was examined by immunolabeling. Origin from the cerebral cortex was determined by fluorescence confocal microscopy. Figure 18 A, 18D), hippocampal CA1 area ( Figure 18 B, 18E) and dentate gyrus ( Figure 18 Anti-type IV collagen markers on thin sections of C, 18F. Scale bar = 30 μM.

[0047] Figure 19 This illustrates areas such as the cerebral cortex, hippocampal CA1, and dentate gyrus (DG) regions of the brains of ApoE- / - mice treated with STZ and subsequently via tail vein injection of PBS mediator or CMP. Figure 18 A series of bar graphs showing the immunofluorescence staining intensity of type IV collagen in the sections shown. Data are expressed as mean ± SEM. *p=0.04; **p=0.01; Mann-Whitney nonparametric t-test.

[0048] Figure 20 It comes from treatment with STZ followed by PBS mediator via tail vein injection. Figure 20 A-20D) or CMP ( Figure 20A series of fluorescence micrographs of mouse brain slices from ApoE- / - mice treated with E-20H. Examination of brain tissue from the CA1 region of the hippocampus using anti-GFAP immunolabeling (all images). Figure 20 A, 20B, 20E and 20F) and the dentate gyrus (DG) region ( Figure 20 Astrocytes from sections of C, 20D, 20G, and 20H were examined for vascular integrity using biotinylate immunolabeling. Figure 20 (B, 20D, 20F, 20H). Markings were determined by fluorescence confocal microscopy. Scale bar = 30 μM.

[0049] Figure 21 It shows things like Figure 20 The hippocampal CA1 cells of ApoE- / - mice shown were treated with STZ and subsequently with either a PBS mediator or CMP via tail vein injection. Figure 21 A) and dentate gyrus (DG) Figure 21 The number of astrocytes present in the slice of region B and the interaction between astrocytes and blood vessels. Figure 21 A(i) and 21B(i): GFAP staining intensity, a measure of the presence of astrocytes. Figure 21 A(ii) and 21B(ii): Quantification of the number of astrocytes in each image (*p=0.02). Figure 21 A(iii) and 21B(iii): Measurement of the shortest distance from the centroid of a given astrocyte to the nearest vessel in each image. ns = not significant, p = 0.07; *p = 0.02; ***p = 0.0004; ****p < 0.0001. Data are presented as mean ± SEM; Mann-Whitney nonparametric t-test. Detailed Implementation

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this invention, preferred methods and materials are described below.

[0051] According to a first aspect, the present invention provides compositions suitable for use in pharmaceuticals for treating or preventing diseases, conditions, structural abnormalities, or injuries in humans or animals requiring treatment or prevention, such as diseases, symptoms, structural abnormalities, or injuries. In some embodiments, the compositions provided by the present invention comprise (a) at least one collagen mimic peptide (CMP) linked to at least one additional therapeutic compound (TC) to form a CMP-TC conjugate, and (b) one or more pharmaceutically suitable carriers. In a related aspect, the present invention provides compositions suitable for use in diagnostic agents for diagnosing or detecting diseases, symptoms, structural abnormalities, or injuries in humans or animals requiring diagnosis. In some embodiments, the compositions provided by the present invention comprise (a) at least one collagen mimic peptide (CMP) linked to at least one diagnostic compound or reagent (DC) to form a CMP-DC conjugate, and (b) one or more pharmaceutically suitable carriers. In other relevant embodiments, the compositions provided by the present invention comprise (a) at least one collagen mimic peptide (CMP) and (b) at least one additional therapeutic compound, wherein the CMP and at least one additional therapeutic compound are mixed in a formulation, or "co-formulated," optionally together with one or more pharmaceutically suitable carriers. In similar embodiments, the compositions provided by the present invention comprise (a) at least one collagen mimic peptide (CMP) and (b) at least one diagnostic compound or reagent, such as a labeling compound or reagent, wherein the CMP and at least one diagnostic compound or reagent are mixed in a formulation, or "co-formulated," optionally together with one or more pharmaceutically suitable carriers, for use in one or more diagnostic methods of the present invention.

[0052] In some embodiments of the invention, the collagen mimic peptide comprises, is substantially composed of, or is composed of a specific tripeptide multimeric repeating amino acid sequence having the sequence (Xaa-Yaa-Gly). n (SEQ ID NO: 417), wherein Xaa is independently selected from the group consisting of proline, 4S-hydroxyproline, fluoroproline, chloroproline, lysine, cysteine ​​and methionine; wherein Yaa is independently selected from the group consisting of proline, 4R-hydroxyproline, fluoroproline, chloroproline, lysine, cysteine ​​and methionine; wherein Gly is a glycine residue; and wherein n is an integer ranging from 1 to 20, such as 3 to 15, 5 to 15 or 5 to 10, and preferably 5, 6, 7, 8, 9 or 10.

[0053] In other embodiments, the present invention provides collagen mimic peptides comprising one or more alternative amino acids in place of at least one amino acid listed in the tripeptide shown in SEQ ID NO: 417, including but not limited to alanine (Ala), glutamine (Gln), glutamic acid (Glu), asparagine (Asn), and aspartic acid (Asp).

[0054] In some embodiments of the invention, the collagen mimic peptide comprises, is substantially composed of, or is composed of an amino acid sequence that is or corresponds to a 21-mer of seven repeats of the three amino acid sequence comprising proline-proline-glycine ((Pro-Pro-Gly)7), i.e., the amino acid sequence: Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Pro-Gly-Pro-Pro-Pro-Gly-Pro-Pro-Pro-Gly-Pro-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 1).

[0055] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or is composed of, a seven-repeated 21-mer comprising three amino acid sequences, wherein hydroxyproline (Hyp), preferably a 4S-hydroxyproline residue, has replaced proline 1 in SEQ ID NO: 1, to produce a seven-repeated sequence of 4S-hydroxyproline-proline-glycine ((Hyp-Pro-Gly)7), i.e., the amino acid sequence: Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 2).

[0056] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or is composed of, a seven-repeated 21-mer comprising three amino acid sequences, wherein Hyp, preferably a 4S-hydroxyproline residue, has replaced proline 2 in SEQ ID NO: 1, to produce a seven-repeated sequence of 4S-hydroxyproline-proline-glycine ((Pro-Hyp-Gly)7), i.e., the amino acid sequence: Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 3).

[0057] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or consists of, a seven-repeated 21-mer comprising three amino acid sequences, wherein fluoroproline (Flp) has replaced proline 1 in SEQ ID NO: 1, producing a seven-repeated sequence of fluoroproline-proline-glycine ((Flp-Pro-Gly)7), namely the amino acid sequence: Flp-Pro-Gly-Flp-Pro-Gly-Flp-Pro-Gly-Flp-Pro-Gly-Flp-Pro-Gly-Flp-Pro-Gly-Flp-Pro-Gly (SEQ ID NO: 4).

[0058] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or consists of, a seven-repeated 21-mer comprising three amino acid sequences, wherein Flp has replaced proline 2 in SEQ ID NO: 1 to produce a seven-repeated sequence of proline-fluoroproline-glycine ((Pro-Flp-Gly)7), namely the amino acid sequence: Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 5).

[0059] In certain other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or is composed of, a seven-repeated 21-mer comprising three amino acid sequences, wherein fluoroproline (Flp) has replaced proline 1 in SEQ ID NO: 1 and Hyp has replaced proline 2 in SEQ ID NO: 1, producing a seven-repeated sequence of fluoroproline-hydroxyproline-glycine ((Flp-Hyp-Gly)7), namely the amino acid sequence: Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 6).

[0060] In a CMP containing Flp, the Flp portion can be in a 4-cis or 4-trans configuration, and is preferably in a 4-cis configuration.

[0061] In some other embodiments of the invention, the collagen mimic peptide comprises, is, consists of, or has the amino acid sequence of a 21-mer consisting of seven repeats of a three-amino acid sequence, wherein chloroproline (Clp) has replaced proline 1 in SEQ ID NO: 1 to produce a seven-repeated sequence of chloroproline-proline-glycine ((Clp-Pro-Gly)7), namely the amino acid sequence: Clp-Pro-Gly-Clp-Pro-Gly-Clp-Pro-Gly-Clp-Pro-Gly-Clp-Pro-Gly-Clp-Pro-Gly-Clp-Pro-Gly (SEQ ID NO: 7).

[0062] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting of, or having, a seven-repeated 21-mer comprising three amino acid sequences, wherein chloroproline (Clp) has replaced proline 2 in SEQ ID NO: 1, producing a seven-repeated sequence of proline-chloroproline-glycine ((Pro-Clp-Gly)7), namely the amino acid sequence: Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 8).

[0063] In certain other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a 21-mer amino acid sequence consisting essentially of, or consists of, a seven-repeated 21-mer comprising three amino acid sequences, wherein Clp has replaced proline 1 in SEQ ID NO: 1 and Hyp has replaced proline 2 in SEQ ID NO: 1, producing a seven-repeated sequence of chloroproline-hydroxyproline-glycine ((Clp-Hyp-Gly)7), namely the amino acid sequence: Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 9).

[0064] In a CMP containing Clp, the Clp moiety can be in a 4-cis or 4-trans configuration, and is preferably in a 4-cis configuration.

[0065] In certain other embodiments of the invention, the collagen mimic peptide may comprise, consist of, or have the amino acid sequence of a 21-mer corresponding to any one of SEQ ID NO:1-9, wherein at least one cysteine ​​(Cys) residue has substituted for at least one proline residue in SEQ ID NO:1, at least one hydroxyproline residue in SEQ ID NO:2-3 and 6, at least one fluoroproline residue in SEQ ID NO:4-6, or at least one chloroproline residue in SEQ ID NO:7-9, thereby producing, for example, the following sequence:

[0066] Pro-Pro-Gly-Pro-Pro-Gly-Cys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 10);

[0067] Hyp-Pro-Gly-Hyp-Pro-Gly-Cys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 11);

[0068] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Cys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 12);

[0069] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 13);

[0070] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 14);

[0071] Cys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 15);

[0072] Pro-Cys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 16);

[0073] Pro-Pro-Gly-Cys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 17);

[0074] Pro-Pro-Gly-Pro-Cys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 18);

[0075] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Cys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 19);

[0076] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Cys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 20);

[0077] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Cys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 21);

[0078] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Cys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 22);

[0079] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Cys-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 23);

[0080] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Cys-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 24);

[0081] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Cys-Gly-Cys-Pro-Gly (SEQ ID NO: 25);

[0082] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Cys-Pro-Gly (SEQ ID NO: 26);

[0083] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Cys-Gly (SEQ ID NO: 27);

[0084] Cys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 28);

[0085] Hyp-Cys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 29);

[0086] Hyp-Pro-Gly-Cys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 30);

[0087] Hyp-Pro-Gly-Hyp-Cys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 31);

[0088] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Cys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 32);

[0089] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Cys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 33);

[0090] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Cys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 34);

[0091] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Cys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 35);

[0092] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Cys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 36);

[0093] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Cys-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 37);

[0094] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Cys-Gly-Hyp-Pro-Gly (SEQ ID NO: 38);

[0095] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Cys-Pro-Gly (SEQ ID NO: 39);

[0096] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Cys-Gly (SEQ ID NO: 40);

[0097] Cys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 41);

[0098] Pro-Cys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 42);

[0099] Pro-Hyp-Gly-Cys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 43);

[0100] Pro-Hyp-Gly-Pro-Cys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 44);

[0101] Pro-Hyp-Gly-Pro-Hyp-Gly-Cys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 45);

[0102] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Cys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 46);

[0103] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Cys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 47);

[0104] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Cys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 48);

[0105] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Cys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 49);

[0106] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Cys-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 50);

[0107] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Cys-Gly-Pro-Hyp-Gly (SEQ ID NO: 51);

[0108] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Cys-Hyp-Gly (SEQ ID NO: 52);

[0109] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Cys-Gly (SEQ ID NO: 53);

[0110] Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 54);

[0111] Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 55);

[0112] Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 56);

[0113] Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 57);

[0114] Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 58);

[0115] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 59);

[0116] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 60);

[0117] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 61);

[0118] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 62);

[0119] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 63);

[0120] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly (SEQ ID NO: 64);

[0121] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly (SEQ ID NO: 65);

[0122] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly (SEQ ID NO: 66);

[0123] Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 67);

[0124] Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 68);

[0125] Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 69);

[0126] Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 70);

[0127] Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 71);

[0128] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 72);

[0129] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 73);

[0130] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 74);

[0131] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 75);

[0132] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 76);

[0133] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 77);

[0134] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly-Pro-Flp-Gly (SEQ ID NO: 78);

[0135] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Cys-Flp-Gly (SEQ ID NO: 79);

[0136] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Cys-Gly (SEQ ID NO: 80);

[0137] Cys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 81);

[0138] Flp-Cys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 82);

[0139] Flp-Hyp-Gly-Cys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 83);

[0140] Flp-Hyp-Gly-Flp-Cys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 84);

[0141] Flp-Hyp-Gly-Flp-Hyp-Gly-Cys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 85);

[0142] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Cys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 86);

[0143] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Cys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 87);

[0144] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Cys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 88);

[0145] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Cys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 89);

[0146] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Cys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 90);

[0147] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Cys-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 91);

[0148] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Cys-Gly-Flp-Hyp-Gly (SEQ ID NO: 92);

[0149] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Cys-Hyp-Gly (SEQ ID NO: 93);

[0150] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Cys-Gly (SEQ ID NO: 94);

[0151] Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 95);

[0152] Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 96);

[0153] Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 97);

[0154] Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 98);

[0155] Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 99);

[0156] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 100);

[0157] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 101);

[0158] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 102);

[0159] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 103);

[0160] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 104);

[0161] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly (SEQ ID NO: 105);

[0162] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly (SEQ ID NO: 106);

[0163] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly (SEQ ID NO: 107);

[0164] Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 108);

[0165] Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 109);

[0166] Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 110);

[0167] Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 111);

[0168] Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 112);

[0169] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 113);

[0170] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 114);

[0171] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 115);

[0172] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 116);

[0173] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 117);

[0174] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 118);

[0175] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly-Pro-Clp-Gly (SEQ ID NO: 119);

[0176] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Cys-Clp-Gly (SEQ ID NO: 120);

[0177] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Cys-Gly (SEQ ID NO: 121);

[0178] Cys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 122);

[0179] Clp-Cys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 123);

[0180] Clp-Hyp-Gly-Cys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 124);

[0181] Clp-Hyp-Gly-Clp-Cys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 125);

[0182] Clp-Hyp-Gly-Clp-Hyp-Gly-Cys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 126);

[0183] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Cys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 127);

[0184] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Cys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 128);

[0185] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Cys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 129);

[0186] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Cys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 130);

[0187] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Cys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 131);

[0188] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Cys-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 132);

[0189] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Cys-Gly-Clp-Hyp-Gly (SEQ ID NO: 133);

[0190] and

[0191] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Cys-Gly (SEQ ID NO: 135).

[0192] In certain other embodiments of the invention, the collagen mimic peptide may comprise, consist of, or have the amino acid sequence of a 21-mer corresponding to any one of SEQ ID NO: 1-9, wherein at least one methionine (Met) residue has substituted for at least one proline residue of SEQ ID NO: 1, at least one hydroxyproline residue of SEQ ID NO: 2-3 and 6, at least one fluoroproline residue of SEQ ID NO: 4-6, or at least one chloroproline residue of SEQ ID NO: 7-9, thereby producing, for example, the following sequence:

[0193] Pro-Pro-Gly-Pro-Pro-Gly-Met-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 136);

[0194] Hyp-Pro-Gly-Hyp-Pro-Gly-Met-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 137);

[0195] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Met-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 138);

[0196] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 139);

[0197] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 140);

[0198] Met-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 141);

[0199] Pro-Met-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 142);

[0200] Pro-Pro-Gly-Met-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 143);

[0201] Pro-Pro-Gly-Pro-Met-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 144);

[0202] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Met-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 145);

[0203] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Met-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 146);

[0204] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Met-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 147);

[0205] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Met-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 148);

[0206] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Met-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 149);

[0207] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Met-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 150);

[0208] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Met-Gly-Pro-Pro-Gly (SEQ ID NO: 151);

[0209] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Met-Pro-Gly (SEQ ID NO: 152);

[0210] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Met-Gly (SEQ ID NO: 153);

[0211] Met-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 154);

[0212] Hyp-Met-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 155);

[0213] Hyp-Pro-Gly-Met-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 156);

[0214] Hyp-Pro-Gly-Hyp-Met-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 157);

[0215] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Met-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 158);

[0216] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Met-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 159);

[0217] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Met-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 160);

[0218] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Met-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 161);

[0219] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Met-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 162);

[0220] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Met-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 163);

[0221] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Met-Gly-Hyp-Pro-Gly (SEQ ID NO: 164);

[0222] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Met-Pro-Gly (SEQ ID NO: 165);

[0223] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Met-Gly (SEQ ID NO: 166);

[0224] Met-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 167);

[0225] Pro-Met-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 168);

[0226] Pro-Hyp-Gly-Met-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 169);

[0227] Pro-Hyp-Gly-Pro-Met-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 170);

[0228] Pro-Hyp-Gly-Pro-Hyp-Gly-Met-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 171);

[0229] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Met-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 172);

[0230] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Met-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 173);

[0231] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Met-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 174);

[0232] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Met-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 175);

[0233] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Met-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 176);

[0234] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Met-Gly-Pro-Hyp-Gly (SEQ ID NO: 177);

[0235] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Met-Hyp-Gly (SEQ ID NO: 178);

[0236] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Met-Gly (SEQ ID NO: 179);

[0237] Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 180);

[0238] Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 181);

[0239] Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 182);

[0240] Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 183);

[0241] Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 184);

[0242] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 185);

[0243] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 186);

[0244] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 187);

[0245] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 188);

[0246] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 189);

[0247] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly (SEQ ID NO: 190);

[0248] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly (SEQ ID NO: 191);

[0249] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly (SEQ ID NO: 192);

[0250] Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 193);

[0251] Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 194);

[0252] Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 195);

[0253] Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 196);

[0254] Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 197);

[0255] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 198);

[0256] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 199);

[0257] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 200);

[0258] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 201);

[0259] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 202);

[0260] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 203);

[0261] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly-Pro-Flp-Gly (SEQ ID NO: 204);

[0262] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Met-Flp-Gly (SEQ ID NO: 205);

[0263] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Met-Gly (SEQ ID NO: 206);

[0264] Met-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 207);

[0265] Flp-Met-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 208);

[0266] Flp-Hyp-Gly-Met-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 209);

[0267] Flp-Hyp-Gly-Flp-Met-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 210);

[0268] Flp-Hyp-Gly-Flp-Hyp-Gly-Met-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 211);

[0269] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Met-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 212);

[0270] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Met-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 213);

[0271] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Met-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 214);

[0272] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Met-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 215);

[0273] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Met-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 216);

[0274] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Met-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 217);

[0275] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Met-Gly-Flp-Hyp-Gly (SEQ ID NO: 218);

[0276] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Met-Hyp-Gly (SEQ ID NO: 219);

[0277] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Met-Gly (SEQ ID NO: 220);

[0278] Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 221);

[0279] Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 222);

[0280] Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 223);

[0281] Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 224);

[0282] Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 225);

[0283] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 226);

[0284] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 227);

[0285] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 228);

[0286] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 229);

[0287] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 230);

[0288] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly (SEQ ID NO: 231);

[0289] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly (SEQ ID NO: 232);

[0290] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly (SEQ ID NO: 233);

[0291] Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 234);

[0292] Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 235);

[0293] Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 236);

[0294] Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 237);

[0295] Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 238);

[0296] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 239);

[0297] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 240);

[0298] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 241);

[0299] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 242);

[0300] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 243);

[0301] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 244);

[0302] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly-Pro-Clp-Gly (SEQ ID NO: 245);

[0303] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Met-Clp-Gly (SEQ ID NO: 246);

[0304] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Met-Gly (SEQ ID NO: 247);

[0305] Met-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 248);

[0306] Clp-Met-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 249);

[0307] Clp-Hyp-Gly-Met-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 250);

[0308] Clp-Hyp-Gly-Clp-Met-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 251);

[0309] Clp-Hyp-Gly-Clp-Hyp-Gly-Met-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 252);

[0310] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Met-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 253);

[0311] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Met-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 254);

[0312] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Met-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 255);

[0313] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Met-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 256);

[0314] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Met-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 257);

[0315] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Met-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 258);

[0316] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Met-Gly-Clp-Hyp-Gly (SEQ ID NO: 259);

[0317] and

[0318] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Met-Gly (SEQ ID NO: 261).

[0319] In certain other embodiments of the invention, the collagen mimic peptide may comprise, consist of, or have the amino acid sequence of a 21-mer corresponding to any one of SEQ ID NO:1-9, wherein at least one lysine residue has substituted for at least one proline residue in SEQ ID NO:1, at least one hydroxyproline residue in SEQ ID NO:2-3 and 6, at least one fluoroproline residue in SEQ ID NO:4-6, or at least one chloroproline residue in SEQ ID NO:7-9, thereby producing, for example, the following sequence:

[0320] Pro-Pro-Gly-Pro-Pro-Gly-Lys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 262);

[0321] Hyp-Pro-Gly-Hyp-Pro-Gly-Lys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 263);

[0322] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Lys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 264);

[0323] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 265);

[0324] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 266);

[0325] Lys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 267);

[0326] Pro-Lys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 268);

[0327] Pro-Pro-Gly-Lys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 269);

[0328] Pro-Pro-Gly-Pro-Lys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 270);

[0329] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Lys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 271);

[0330] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Lys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 272);

[0331] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Lys-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 273);

[0332] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Lys-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 274);

[0333] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Lys-Gly-Pro-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 275);

[0334] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Lys-Pro-Gly-Pro-Pro-Gly (SEQ ID NO: 276);

[0335] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Lys-Gly-Pro-Pro-Gly (SEQ ID NO: 277);

[0336] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Lys-Pro-Gly (SEQ ID NO: 278);

[0337] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Lys-Gly (SEQ ID NO: 279);

[0338] Lys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 280);

[0339] Hyp-Lys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 281);

[0340] Hyp-Pro-Gly-Lys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 282);

[0341] Hyp-Pro-Gly-Hyp-Lys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 283);

[0342] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Lys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 284);

[0343] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Lys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 285);

[0344] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Lys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 286);

[0345] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Lys-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 287);

[0346] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Lys-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 288);

[0347] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Lys-Pro-Gly-Hyp-Pro-Gly (SEQ ID NO: 289);

[0348] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Lys-Gly-Hyp-Pro-Gly (SEQ ID NO: 290);

[0349] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Lys-Pro-Gly (SEQ ID NO: 291);

[0350] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Lys-Gly (SEQ ID NO: 292);

[0351] Lys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 293);

[0352] Pro-Lys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 294);

[0353] Pro-Hyp-Gly-Lys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 295);

[0354] Pro-Hyp-Gly-Pro-Lys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 296);

[0355] Pro-Hyp-Gly-Pro-Hyp-Gly-Lys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 297);

[0356] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Lys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 298);

[0357] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Lys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 299);

[0358] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Lys-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 300);

[0359] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Lys-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 301);

[0360] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Lys-Hyp-Gly-Pro-Hyp-Gly (SEQ ID NO: 302);

[0361] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Lys-Gly-Pro-Hyp-Gly (SEQ ID NO: 303);

[0362] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Lys-Hyp-Gly (SEQ ID NO: 304);

[0363] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Lys-Gly (SEQ ID NO: 305);

[0364] Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 306);

[0365] Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 307);

[0366] Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 308);

[0367] Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 309);

[0368] Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 310);

[0369] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 311);

[0370] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 312);

[0371] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 313);

[0372] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 314);

[0373] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 315);

[0374] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly (SEQ ID NO: 316);

[0375] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly (SEQ ID NO: 317);

[0376] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly (SEQ ID NO: 318);

[0377] Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 319);

[0378] Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 320);

[0379] Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 321);

[0380] Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 322);

[0381] Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 323);

[0382] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 324);

[0383] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 325);

[0384] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 326);

[0385] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 327);

[0386] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 328);

[0387] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly-Pro-Flp-Gly (SEQ ID NO: 329);

[0388] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly-Pro-Flp-Gly (SEQ ID NO: 330);

[0389] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Lys-Flp-Gly (SEQ ID NO: 331);

[0390] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Lys-Gly (SEQ ID NO: 332);

[0391] Lys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 333);

[0392] Flp-Lys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 334);

[0393] Flp-Hyp-Gly-Lys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 335);

[0394] Flp-Hyp-Gly-Flp-Lys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 336);

[0395] Flp-Hyp-Gly-Flp-Hyp-Gly-Lys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 337);

[0396] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Lys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 338);

[0397] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Lys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 339);

[0398] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Lys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 340);

[0399] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Lys-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 341);

[0400] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Lys-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 342);

[0401] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Lys-Hyp-Gly-Flp-Hyp-Gly (SEQ ID NO: 343);

[0402] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Lys-Gly-Flp-Hyp-Gly (SEQ ID NO: 344);

[0403] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Lys-Hyp-Gly (SEQ ID NO: 345);

[0404] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Lys-Gly (SEQ ID NO: 346);

[0405] Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 347);

[0406] Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 348);

[0407] Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 349);

[0408] Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 350);

[0409] Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 351);

[0410] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 352);

[0411] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 353);

[0412] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 354);

[0413] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 355);

[0414] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 356);

[0415] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly (SEQ ID NO: 357);

[0416] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly (SEQ ID NO: 358);

[0417] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly (SEQ ID NO: 359);

[0418] Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 360);

[0419] Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 361);

[0420] Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 362);

[0421] Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 363);

[0422] Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 364);

[0423] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 365);

[0424] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 366);

[0425] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 367);

[0426] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 368);

[0427] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 369);

[0428] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly-Pro-Clp-Gly (SEQ ID NO: 370);

[0429] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly-Pro-Clp-Gly (SEQ ID NO: 371);

[0430] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Lys-Clp-Gly (SEQ ID NO: 372);

[0431] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Lys-Gly (SEQ ID NO: 373);

[0432] Lys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 374);

[0433] Clp-Lys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 375);

[0434] Clp-Hyp-Gly-Lys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 376);

[0435] Clp-Hyp-Gly-Clp-Lys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 377);

[0436] Clp-Hyp-Gly-Clp-Hyp-Gly-Lys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 378);

[0437] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Lys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 379);

[0438] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Lys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 380);

[0439] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Lys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 381);

[0440] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Lys-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 382);

[0441] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Lys-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 383);

[0442] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Lys-Hyp-Gly-Clp-Hyp-Gly (SEQ ID NO: 384);

[0443] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Lys-Gly-Clp-Hyp-Gly (SEQ ID NO: 385);

[0444] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Lys-Hyp-Gly (SEQ ID NO: 386); and

[0445] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Lys-Gly (SEQ ID NO: 387).

[0446] Another suitable CMP for use in accordance with the present invention is a CMP having or comprising the sequence Hyp-Flp-Gly-Hyp-Flp-Gly-Hyp-Flp-Gly-Hyp-Flp-Gly-Hyp-Flp-Gly-Flp-Gly-Hyp-Flp-Gly (SEQ ID NO: 388).

[0447] Other suitable CMPs used according to the present invention are CMPs having or comprising the following sequences: Gly3-(Pro-Hyp-Gly)6 (SEQ ID NO:397), Gly3-(Pro-Flp-Gly)6 (SEQ ID NO:398), Gly3-(Pro-Hyp-Gly)7 (SEQ ID NO:399), Gly3-(Pro-Flp-Gly)7 (SEQ ID NO:400), Gly3-(Pro-Hyp-Gly)8 (SEQ ID NO:401), Gly3-(Pro-Flp-Gly)8 (SEQ ID NO:402), Gly3-(Pro-Hyp-Gly)9 (SEQ ID NO:403), Gly3-(Pro-Flp-Gly)9 (SEQ ID NO:404), (Pro-Hyp-Gly)6-Tyr (SEQ ID NO:397 ...6 (SEQ ID NO:398), Gly3-(Pro-Hyp-Gly)7 (SEQ ID NO:399), Gly3-(Pro-Flp-Gly)7 (SEQ ID NO:400), Gly3-(Pro-Hyp-Gly)8 (SEQ ID NO:401), Gly3-(Pro-Flp-Gly)9 (SEQ ID NO:402), (Pro-Hyp-Gly)6-Tyr (SEQ ID NO:403), Gly3-(Pro-Flp-Gly)9 (SEQ ID NO:404), (Pro-Hyp-Gly)6-Tyr (SEQ ID NO:397), Gly3-(Pro-Flp-Gly)9 (SEQ ID NO:404), (Pro-Hyp-Gly NO:405), (Pro-Flp-Gly)6-Tyr (SEQ ID NO:406), (Pro-Hyp-Gly)7-Tyr (SEQ ID NO:407), (Pro-Flp-Gly)7-Tyr (SEQ ID NO:408), (Pro-Hyp-Gly)8-Tyr (SEQ ID NO:409), (Pro-Flp-Gly)8-Tyr (SEQ ID NO:410), Cys-(Pro-Hyp-Gly)3 (SEQ ID NO:411), Cys-(Pro-Flp-Gly)3 (SEQ ID NO:412), Cys-(Pro-Hyp-Gly)5 (SEQ ID NO:413), Cys-(Pro-Flp-Gly)5 (SEQ ID NO:414), Cys-(Pro-Hyp-Gly)7 (SEQ ID NO:415) and Cys-(Pro-Flp-Gly)7 (SEQ ID NO:416), as well as other similar CMPs, may be suitable for use as in vitro and in vivo collagen modifiers in therapeutic and / or diagnostic methods (see, for example, U.S. Patent Nos. 8,283,414, 8,883,964 and 10,632,168, and U.S. Patent Publication No. US 2020 / 0353056 A1, the disclosures of all of which are incorporated herein by reference in their entirety).

[0448] In some other embodiments of the invention, the collagen mimic peptide comprises, is or corresponds to, a seven-repeated 21-mer amino acid sequence comprising three amino acid sequences, is substantially composed of, or is composed of, the amino acid sequence wherein Ala has replaced proline 2 in SEQ ID NO: 1, thereby producing a seven-repeated sequence of proline-alanine-glycine ((Pro-Ala-Gly)7), namely the amino acid sequence: Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly (SEQ ID NO: 418). In other embodiments, the collagen mimic peptide comprises, is substantially composed of, or is composed of a 21-mer amino acid sequence that is or corresponds to a seven-repeated 21-mer comprising three amino acid sequences, wherein Hyp or Flp has replaced proline 1 in SEQ ID NO: 418 to produce a seven-repeated sequence of proline-alanine-glycine ((Pro-Ala-Gly)7), i.e., the amino acid sequences: Hyp-Ala-Gly-Hyp-Ala-Gly-Hyp-Ala-Gly-Hyp-Ala-Gly-Hyp-Ala-Gly-Hyp-Ala-Gly (SEQ ID NO: 419) and Flp-Ala-Gly-Flp-Ala-Gly-Flp-Ala-Gly-Flp-Ala-Gly-Flp-Ala-Gly-Flp-Ala-Gly-Flp-Ala-Gly (SEQ ID NO:420).

[0449] Other suitable CMPs for use according to the present invention are CMPs having or comprising the following sequences: (Pro-Gly-Glu)7 (SEQ ID NO: 421), (Pro-Gly-Gln)7 (SEQ ID NO: 422), (Pro-Gly-Pro)7 (SEQ ID NO: 423), (Hyp-Gly-Glu)7 (SEQ ID NO: 424), (Hyp-Gly-Gln)7 (SEQ ID NO: 425), (Flp-Gly-Glu)7 (SEQ ID NO: 426), (Flp-Gly-Gln)7 (SEQ ID NO: 427), (Hyp-Gly-Pro)7 (SEQ ID NO: 428), (Hyp-Gly-Gln)7 (SEQ ID NO: 429), (Hyp-Gly-Glu)7 (SEQ ID NO: 430), (Pro-Gly-Asp)7 (SEQ ID NO: 431), (Pro-Gly-Asn)7 (SEQ ID NO: 432), (Lys-Gly-Gln)7 (SEQ ID NO: 433), (Lys-Gly-Glu)7 (SEQ ID NO: 434), (Hyp-Ala-Gly)7 (SEQ ID NO: 435), (Flp-Ala-Gly)7 (SEQ ID NO: 436), (Hyp-Gly-Glu)7 (SEQ ID NO: 437), (Hyp-Gly-Gln)7 (SEQ ID NO: 438), (Flp-Gly-Glu)7 (SEQ ID NO: 439), (Flp-Gly-Gln)7 (SEQ ID NO: 440), (Hyp-Gly-Asp)7 (SEQ ID NO: 441), (Hyp-Gly-Asn)7 (SEQ ID NO: 442), (Flp-Gly-Asp)7 (SEQ ID NO: 443), (Flp-Gly-Asn)7 (SEQ ID NO: 444), (Hyp-Gly-Pro)7 (SEQ ID NO: 445), (Flp-Gly-Pro)7 (SEQ ID NO: 446), (Pro-Gly-Hyp)7 (SEQ ID NO: 447), (Flp-Gly-Hyp)7 (SEQ ID NO: 448), (Hyp-Gly-Hyp)7 (SEQ ID NO: 449), (Hyp-Gly-Flp)7 (SEQ ID NO: 450), (Pro-Gly-Flp)7(SEQ ID NO: 451), (Flp-Gly-Flp)7 (SEQ ID NO: 452), and other similar CMPs, which contain one or more additional or substituted amino acids (including one or more cysteine ​​residues, one or more methionine residues and / or one or more lysine residues) inserted into or substituted at the positions indicated by cysteine ​​in SEQ ID No: 10-135, methionine in SEQ ID No: 136-251, and lysine in SEQ ID No: 262-387.

[0450] In some other embodiments, any of the aforementioned CMPs may optionally have one or more cysteine ​​residues, one or more methionine residues, and / or one or more lysine residues attached to the N-terminus or C-terminus of the amino acid sequence, or attached to both ends. Non-limiting examples of such CMPs include:

[0451] SEQ ID NO: 1, wherein the cysteine ​​residue is attached to the N-terminus, i.e., Cys-((Pro-Pro-Gly)7) (SEQ ID NO: 453), attached to the C-terminus, i.e. (Pro-Pro-Gly)7-Cys (SEQ ID NO: 454), or attached to both ends, i.e. Cys-((Pro-Pro-Gly)7)-Cys (SEQ ID NO: 455);

[0452] SEQ ID NO: 6, wherein the cysteine ​​residue is attached to the N-terminus, i.e., Cys-((Flp-Hyp-Gly)7) (SEQ ID NO: 456), attached to the C-terminus, i.e. (Flp-Hyp-Gly)7-Cys (SEQ ID NO: 457), or attached to both ends, i.e. Cys-((Flp-Hyp-Gly)7)-Cys (SEQ ID NO: 458);

[0453] SEQ ID NO: 1, wherein the methionine residue is attached to the N-terminus, i.e., Met-((Pro-Pro-Gly)7) (SEQ ID NO: 459), or to the C-terminus, i.e. (Pro-Pro-Gly)7-Met (SEQ ID NO: 460), or to both ends, i.e. Met-((Pro-Pro-Gly)7)-Met (SEQ ID NO: 461);

[0454] SEQ ID NO: 6, wherein the methionine residue is attached to the N-terminus, i.e., Met-((Flp-Hyp-Gly)7) (SEQ ID NO: 462), attached to the C-terminus, i.e. (Flp-Hyp-Gly)7-Met (SEQ ID NO: 463), or attached to both ends, i.e. Met-((Flp-Hyp-Gly)7)-Met (SEQ ID NO: 464);

[0455] SEQ ID NO: 1, wherein the lysine residue is attached to the N-terminus, namely Lys-((Pro-Pro-Gly)7) (SEQ ID NO:465), or to the C-terminus, namely (Pro-Pro-Gly)7-Lys (SEQ ID NO:466), or to both ends, namely Lys-((Pro-Pro-Gly)7)-Lys (SEQ ID NO:467);

[0456] SEQ ID NO: 6, wherein the lysine residue is attached to the N-terminus, i.e., Lys-((Flp-Hyp-Gly)7) (SEQ ID NO: 468), attached to the C-terminus, i.e. (Flp-Hyp-Gly)7-Lys (SEQ ID NO: 469), or attached to both ends, i.e. Lys-((Flp-Hyp-Gly)7)-Lys (SEQ ID NO: 470).

[0457] In other embodiments, the sequences listed in SEQ ID Nos. 453-458 may have one or more methionine or lysine residues replacing one or more cysteine ​​residues. In other embodiments, the sequences listed in SEQ ID Nos. 459-464 may have one or more methionine or lysine residues replacing one or more methionine residues. In other embodiments, the sequences listed in SEQ ID Nos. 465-470 may have one or more lysine residues replacing one or more methionine or cysteine ​​residues.

[0458] Of course, based on the knowledge in this art and the teachings herein, those skilled in the art will understand that such CMPs may contain two or more cysteine, methionine, and / or lysine residues, wherein at least one additional cysteine, methionine, and / or lysine residue, or any combination thereof, may substitute for at least one proline residue, at least one hydroxyproline residue, at least one fluoroproline residue, and / or at least one chloroproline residue in any of the aforementioned CMP sequences containing at least one proline, at least one hydroxyproline, at least one fluoroproline residue, and / or at least one chloroproline residue. Those skilled in the art, based on the teachings herein and the information readily available in the art, should also understand that other combinations of amino acid substitutions are possible and fall within the scope of this invention.

[0459] The CMP described herein is applicable to a variety of purposes. For example, as further described elsewhere herein, CMP can be used for a variety of therapeutic or preventative applications by direct application to or introduction into the human or animal body, particularly at sites of collagen damage or potential collagen damage. In this case, the CMP described herein will directly target the site of collagen damage, anneal with the damaged collagen chains, stabilize the collagen structure to resist further damage, and in some cases, reform the natural collagen triple helix at the site of collagen damage. Such applications help promote the repair and strengthening of damaged collagen in injured or potentially injured or damaged sites such as wounds, diseases, structural abnormalities, or conditions (e.g., scarring, wrinkle formation, etc.) involving the skin, tendons, ligaments, cartilage, bone, and other collagen-containing structures and organs. The CMP described herein is also used to provide biocompatible coatings for certain medical devices to promote the healing of injuries and conditions in body areas, wherein such devices are used to treat or prevent certain diseases, conditions, and structural abnormalities or injuries in humans and animals, particularly those involving the destruction of collagen and / or collagen-containing structures in body areas. The CMP described herein is also used to provide unique delivery media suitable for delivering various therapeutic compounds, compositions, and drugs to sites of diseases, conditions, and structural abnormalities or injuries in humans and animals, particularly for treating, preventing, or improving diseases, conditions, medical conditions, and structural abnormalities or injuries caused by collagen destruction that are associated with or co-located to the site of the disease, condition, or injury. In another embodiment, the CMP described herein can be used to provide diagnostic agents suitable for diagnosing or detecting diseases, conditions, structural abnormalities, or injuries in humans and animals. In some of these respects, CMP can be formulated or directly or indirectly conjugated with one or more suitable diagnostic compounds, pharmaceuticals, markers, etc. (see, for example, U.S. Patent Nos. 8,283,414, 8,883,964, and 10,632,168, and U.S. Patent Publication No. US2020 / 0353056 A1, the disclosures of all these patents being incorporated herein by reference in their entirety). Other suitable uses of CMP described herein and used in certain aspects of this invention will be apparent to those skilled in the art based on the disclosures herein and information readily available in the art.

[0460] In some embodiments, the CMPs described herein are suitable for forming films, sheets, membranes, or gels comprising one or more CMPs, in a form suitable for introduction or implantation into humans or animals for therapeutic, preventative, or diagnostic applications, such as those described herein, as well as other applications familiar to those skilled in the art. For example, films, sheets, membranes, spheres, nanoparticles, or gels can be formed from one or more CMP solutions described herein using methods such as those described in U.S. Patent Nos. 6,197,934, 6,448,378, and 9,289,396; the disclosures of all such patents are incorporated herein by reference in their entirety. Alternatively, films, sheets, membranes, spheres, nanoparticles, or gels may be formed from other materials, such as determinated collagen (see U.S. Patent Nos. 6,197,934; 6,448,378; and 9,289,396), copolymers of poly(lactic acid) and poly(glycolic acid) (PLGA) (see Bala, I., et al., Crit. Rev. Ther. Drug Carrier Syst. 21(5):387-422 (2004)), and other materials known to those skilled in the art (see, for example, Kumar, V., et al., eds., “Polymer Gels: Perspectives and Applications”, ISBN 978-981-10-6079-3, Singapore: Springer). (2018)), and one or more CMPs can be suitably incorporated into the solution during the formation of such films, sheets, membranes, spheres, nanoparticles, gels, etc., at concentrations of about 1%-99%, about 2%-95%, about 3%-90%, about 4%-90%, about 5%-90%, about 10%-90%, about 15%-90%, about 20%-90%, about 25%-90%, about 25%-85%, about 25%-75%, about 25%-50%, about 35%-50%, etc. Based on the teachings herein and information readily available to those skilled in the art, it is readily apparent that suitable other amounts or concentrations of the CMPs described herein can be suitably included in the solution during the formation of films, sheets, membranes, spheres, nanoparticles, gels, etc. In some such embodiments, one or more therapeutic compounds described herein and / or one or more CMP-TC conjugates described herein can be suitably incorporated into the solution forming the films, sheets, membranes, spheres, nanoparticles, gels, etc.Alternatively, in a relevant context, once one or more films, sheets, membranes, spheres, nanoparticles, gels, etc., are formed as described above, they can be treated or coated with one or more CMP and / or CMP-TC conjugates described herein by immersing the film, sheet, membrane, sphere, nanoparticle, gel, etc., in a solution, particularly a buffered aqueous solution, containing a suitable amount or concentration (such as those described herein) of one or more of the CMP or CMP-TC conjugates described herein, and then drying the film, sheet, membrane, etc., prior to use in therapeutic, preventive, or diagnostic methods (such as those described herein).

[0461] CMP connection / joining

[0462] In certain embodiments of the invention, the CMP described herein is suitably linked or conjugated to one or more therapeutic or diagnostic compounds to produce CMP conjugated compounds. In such embodiments of the invention, the CMP-therapeutic compound or CMP-diagnostic compound conjugated compound may subsequently be introduced into the human or animal body in a method of treating and / or preventing and / or diagnosing certain diseases, conditions, and structural abnormalities in a human or animal. Therefore, in some embodiments, the invention also provides the use of linking or conjugating the CMP described herein to one or more therapeutic compounds to produce conjugated CMPs, compositions comprising such conjugated CMPs (which may optionally contain one or more additional therapeutic or pharmaceutically active ingredients), methods of producing such conjugations, and methods of using such conjugations and compositions to treat, prevent, and diagnose various diseases, conditions, and medical conditions in humans and animals.

[0463] According to this aspect of the invention, a conjugate of CMP with at least one therapeutic compound (described herein as a “CMP-TC conjugate”) will comprise at least one CMP described herein, which is linked to at least one therapeutic compound to form the CMP-TC conjugate. CMPs applicable to this aspect of the invention include any of those described herein, including CMPs having an amino acid sequence corresponding to any one of SEQ ID NO: 1-387, and particularly wherein such CMPs have an amino acid sequence corresponding to any one of SEQ ID NO: 1-14, 66-94, 107-135, 136-140, 192-220, 233-261, 260-264, 280, 281, 293, 294, 306, 307, 318-346, 347, 348, 359-388, and 397-416, and more particularly, CMPs having an amino acid sequence corresponding to SEQ ID NO: 10-27, 81-94, 122-135, 207-220, 248-261, 333-346, 374-388, and 397-416. Based on the teachings contained herein, other suitable CMP sequences will be readily apparent to those skilled in the art. For example, CMPs having at least one, and in some cases more than one, cysteine, methionine, or lysine residue substituted for at least one of SEQ ID NO: 1-9, and in some cases more than one proline, hydroxyproline, fluoroproline, or chloroproline residue will be particularly suitable for the production of the CMP-TC conjugates provided and used in this invention. Examples of such suitable CMPs include CMPs having amino acid sequences corresponding to SEQ ID NO: 10-27, 81-94, 122-135, 207-220, 248-261, 333-346, 374-388, 397-416, and 453-470.

[0464] Based on the teachings herein and the information readily available in the art, those skilled in the art will be familiar with the methods for preparing CMP and CMP-TC described herein and provided and used in this invention. For example, CMP can be synthesized using standard protein / peptide synthesis techniques, such as those described in U.S. Patent Nos. 5,973,112; 7,122,521; and 7,858,741; and U.S. Patent Publication No. US2007 / 0275897 A1, the disclosures of all these patents being incorporated herein by reference in their entirety. CMP synthesis can also be achieved by purchasing commercially produced custom-synthesized CMP, for example, from Bachem (Torrance, CA, USA) and RS Synthesis (Louisville, KY, USA). In other embodiments, CMP synthesis can be achieved using genetic engineering and recombinant expression of CMP from prokaryotic or eukaryotic expression systems (see, for example, Buechter, DD et al., J. Biol. Chem. 278(1):645-650 (2003)).

[0465] In synthesizing the peptides described herein, in some embodiments, it is preferable to use certain stereochemical substitutions for amino acids, particularly if hydroxyproline, fluoroproline, or chloroproline are used:

[0466] (1) If hydroxyproline replaces the proline at the Xaa position of the above-mentioned Xaa-Yaa-Gly trimer, in some embodiments, the hydroxyproline has (2R, 4S) stereochemistry, or cis or trans, preferably cis stereochemistry;

[0467] (2) If hydroxyproline replaces the proline at the Yaa position of the above-mentioned Xaa-Yaa-Gly trimer, in some embodiments, the hydroxyproline has (2R, 4S) stereochemistry, or cis or trans, preferably cis stereochemistry;

[0468] (3) If fluoroproline replaces the proline at the Yaa position of the above-mentioned Xaa-Yaa-Gly trimer, in some embodiments, hydroxyproline has (2R, 4S) stereochemistry, or cis or trans, preferably cis stereochemistry;

[0469] (4) If chloroproline replaces the proline at the Yaa position of the above-mentioned Xaa-Yaa-Gly trimer, in some embodiments, hydroxyproline has (2R, 4S) stereochemistry, or cis or trans, preferably cis stereochemistry.

[0470] Other suitable stereochemistry can be determined empirically without resorting to excessive experimentation and is immediately apparent to those skilled in the art. As mentioned above, certain CMPs provided and used in this invention may contain one or more additional substitutions, such as one or more cysteine ​​residues and / or one or more methionine residues, replacing one or more proline residues in a given CMP polymer. Such substitutions can be suitably achieved by adding these residues to the growing CMP peptide chain during synthesis using standard peptide synthesis methods, such as those described elsewhere herein and those known in the art.

[0471] Once prepared, CMPs are suitable for producing the CMP-TC of the present invention, i.e., the therapeutic or diagnostic composition of the present invention, by linking one or more therapeutic compounds to the CMP. In some embodiments, the CMP-TC of the present invention can be prepared by a method comprising (a) providing a collagen mimic peptide having an amino acid sequence corresponding to any one of SEQ ID NO: 1-470, particularly a CMP having an amino acid sequence corresponding to any one of SEQ ID NO: 1-14, 66-94, 107-135, 136-140, 192-220, 233-261, 260-264, 280, 281, 293, 294, 306, 307, 318-346, 347, 348, 359-388, 397-416, and 453-470, and more particularly having an amino acid sequence corresponding to SEQ ID NO: The CMP comprises: (a) a CMP having an amino acid sequence of 10-27, 81-94, 122-135, 207-220, 248-261, 333-346, 374-388, 397-416, and 453-470; (b) at least one therapeutic or diagnostic compound suitable for conjugation to the CMP; and (c) a therapeutic or diagnostic compound being directly or indirectly attached to the CMP. In some cases, particularly when the therapeutic compound is a small peptide biocompound, the therapeutic compound may be directly attached to the CMP via a peptide bond, for example by simply extending the synthesis of the peptide beyond the carboxyl terminus of the CMP and attaching the amino-terminal amino acid of the therapeutic compound to the carboxyl-terminal amino acid of the CMP via a peptide bond. An example of such a CMP-TC is a peptide conjugate in which a wound-healing peptide, referred to as substance P, having the amino acid sequence Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 471), is attached to the CMP described herein. Examples of such conjugates include, for example:

[0472] Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 472);

[0473] Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Hyp-Pro-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 473);

[0474] Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 474);

[0475] Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Flp-Hyp-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 475);

[0476] Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Clp-Hyp-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 476);

[0477] Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Pro-Flp-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 477); and

[0478] Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Pro-Clp-Gly-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met (SEQ ID NO: 478).

[0479] In other methods of the invention, one or more therapeutic or diagnostic compounds are suitably covalently conjugated to or linked to the CMP via a bond other than a peptide bond (see, for example, U.S. Patent Nos. 3,283,414 and 3,883,964, which are incorporated herein by reference in their entirety). For example, the therapeutic compound can be directly linked to the cysteine ​​or methionine residue on the CMP as described herein by covalently binding a hydroxyl or amino group on an amino acid residue (e.g., a lysine residue) on the therapeutic or diagnostic compound (if it is a biomolecule) to a thiol group on a cysteine ​​or methionine residue on the CMP. Alternatively, if the CMP does not contain a cysteine ​​or methionine residue, one or more therapeutic or diagnostic compounds can be linked to or conjugated to the CMP by a reaction between a hydroxyl or amino group on the CMP and a thiol group on an amino acid residue (e.g., at a cysteine ​​or methionine residue) on the therapeutic or diagnostic compound (if it is a biomolecule). In another alternative conjugation method, the therapeutic compound can be directly linked to the lysine residue on the CMP described herein by covalently binding the therapeutic compound to the amino group on the lysine residue, for example using NHS ester conjugation (see, for example, Mattson, G., et al., Molec. Biol. Rep. 17:167-183 (1993); Grabarek, Z. and Gergely, J., Anal. Biochem. 185:131-135 (1990); Staros, JV, et al., Anal. Biochem. 156:220-2 (1986); Timkovich, R., Anal. Biochem. 79:135-43 (1977)). This direct covalent linking or conjugation between the CMP and the therapeutic / diagnostic compound can be achieved using standard reaction techniques familiar to those skilled in the art of organic chemistry.

[0480] In other embodiments, particularly those where the therapeutic or diagnostic compound is not a biological agent (and therefore does not have a peptide structure or amino acid residues having a group suitable for linking to other residues on cysteine, methionine, lysine, or CMP), such as small molecule organic or inorganic therapeutic or diagnostic compounds, at least one therapeutic or diagnostic compound is indirectly linked to a collagen mimic peptide by means of a linkage. In such embodiments, the linkage has two linkable ends, one end being linked to an amino acid residue on a CMP and suitablely linked to a thiol group on a cysteine ​​or methionine residue or an amino group on a lysine residue, and the other end being linked to a hydroxyl or amino group on the therapeutic or diagnostic compound. For example, in some such embodiments, the linkage comprises at least one polymer chain having a first end and a second end, and the first end of the polymer chain is bound to a thiol group on a cysteine ​​or methionine residue or an amino group on a lysine residue on the collagen mimic peptide, and the opposite end or the second end of the polymer chain is bound to an amino or hydroxyl group on the therapeutic compound. In embodiments where the therapeutic or diagnostic compound is not suitable for direct linking to a biopharmaceutical via peptide synthesis as described elsewhere herein, the second end of the linker may be linked to an amino group on an amino acid residue (such as a lysine residue) of the biotherapeutic or diagnostic compound. Suitable such linking methods are well known to those skilled in the art. For example, one linking method suitable for use according to this aspect of the invention comprises a polymer chain portion having one end (particularly the CMP-binding end) containing a thiol-binding group, such as maleimide, and the other end (particularly the therapeutic or diagnostic compound-binding end) containing an amino-binding group, such as N-hydroxysuccinimide. In some such embodiments, the polymer chain is a linear polyethylene glycol chain containing at least four ethylene glycol monomers, such as four to fifty ethylene glycol monomers, ten to forty ethylene glycol monomers, fifteen to thirty ethylene glycol monomers, fifteen to twenty-five ethylene glycol monomers, twenty to twenty-five ethylene glycol monomers, particularly four, six, eight, twelve, twenty, twenty-two, twenty-three, twenty-four, or twenty-five ethylene glycol monomers. Such connection devices suitable for connecting one or more therapeutic or diagnostic compounds to a CMP via the methods described herein are commercially available, for example (e.g., SM(PEG)6, SM(PEG)8, SM(PEG)12 and SM(PEG)24) from Thermo Fisher Scientific (Waltham, MA).By adjusting the length of the polymer chain, the bioavailability and sustainability of therapeutic or diagnostic compounds in vivo can be modulated—using a longer polymer chain, such as a polymer containing 24 ethylene glycol monomers, will increase the bioavailability of the compound after CMP-TC enters the human or animal body, while using a shorter polymer chain, such as a polymer containing six ethylene glycol monomers, will decrease bioavailability, thereby increasing the sustainability of the therapeutic or diagnostic compound (in other words, resulting in delayed or sustained release). Other conjugates using linear or star-shaped PEG moieties that can be suitably prepared using the CMP of the present invention and used in the therapeutic and diagnostic methods of the present invention are disclosed in U.S. Patent Nos. 8,283,414 and 8,883,964, which are incorporated herein by reference in their entirety. Thus, according to certain aspects of the present invention, at least one therapeutic compound comprises at least one reactive hydroxyl group capable of crosslinking with a collagen-mimicking peptide using a polymer linker.

[0481] According to the invention, other indirect linking methods for conjugating one or more therapeutic or diagnostic compounds to or above a CMP are also suitably used. For example, at least one therapeutic or diagnostic compound may be encapsulated within at least one nanoparticle linked to a collagen mimic peptide via a linking manner as described herein. Alternatively, the collagen mimic peptide may suitably contain at least one biotin moiety, and the therapeutic molecule may suitably contain at least one avidin or streptavidin moiety, with the biotin moiety on the collagen mimic peptide binding to the avidin or streptavidin moiety on the therapeutic or diagnostic compound, thereby linking the collagen mimic peptide to the therapeutic or diagnostic compound. Of course, alternatives are also applicable, wherein the collagen mimic peptide may suitably contain at least one avidin or streptavidin moiety, and the therapeutic or diagnostic compound may suitably contain at least one biotin moiety, with the biotin moiety on at least one therapeutic or diagnostic compound binding to the avidin or streptavidin moiety on the collagen mimic peptide, thereby linking the collagen mimic peptide to the therapeutic compound.

[0482] Therefore, according to certain embodiments of the invention, a therapeutic or diagnostic compound may suitably be directly connected to the CMP described herein. In other embodiments of the invention, one or more therapeutic or diagnostic compounds may be indirectly connected to the CMP described herein, for example, by using spacers, connectors, or bridging portions. It should be understood that whether one or more therapeutic compounds are directly or indirectly connected to the CMP, such connection will result in the formation of a conjugate of the CMP and one or more therapeutic compounds, which may be defined herein as a CMP-TC conjugate.

[0483] Suitable therapeutic or diagnostic compounds for connection or conjugation with CMPs to produce the CMP-TC of the present invention include any compound that has been shown to have specific therapeutic or preventative properties against one or more diseases, conditions, physical conditions, or ailments when introduced into a human or animal suffering from or susceptible to one or more diseases, conditions, physical conditions, or ailments. Any therapeutic or diagnostic compound may be used in the conjugates, compositions, and methods of the present invention, provided that the therapeutic or diagnostic compound can be conjugated or connected to at least one CMP in accordance with the teachings herein. Suitable such therapeutic compounds may be biological or non-biological (e.g., so-called “small molecule”) therapeutic compounds. Suitable compounds include, but are not limited to, steroidal anti-inflammatory drugs (e.g., prednisolone or pharmaceutically acceptable salts thereof, such as prednisolone acetate), nonsteroidal anti-inflammatory drugs (e.g., acetylsalicylic acid, acetaminophen, ibuprofen, naproxen, napafenac, bromfenac, diclofenac, flurbiprofen, ketoprofen, ketorolac, and indene derivatives (e.g., indomethacin, sulinoril, etc.; see, for example, U.S. Patent No. 7,601,874, which is incorporated herein by reference in its entirety, other indene derivatives used as active pharmaceutical ingredients) and their pharmaceutically acceptable salts. Esters and derivatives), local anesthetics (e.g., tetracaine, lidocaine, oxybuprocaine, promecaine, etc.), vitamins or vitamin derivatives or vitamin precursors (e.g., retinol, retinoic acid, retinaldehyde, carotene and other retinoids and retinoid derivatives or precursors; folic acid; α-tocopherol; calciferol; phylloquinone, menadione and other forms, precursors or derivatives of vitamin K, ascorbic acid; etc.), therapeutic enzymes or their therapeutic fragments (e.g., collagenases and serine proteases, or their therapeutically effective fragments), antibiotics (e.g., aminoglycoside antibiotics (e.g., gentamicin)). Sulfonamide antibiotics include tobramycin, paromomycin, kanamycin, neomycin, and amikacin, and their pharmaceutically acceptable salts or esters, such as tobramycin sulfate; fluoroquinolone antibiotics (such as moxifloxacin, gatifloxacin, levofloxacin, gemifloxacin, ciprofloxacin, norfloxacin, and ofloxacin, and their pharmaceutically acceptable salts, esters, or derivatives, such as moxifloxacin hydrochloride, ciprofloxacin hydrochloride, and gatifloxacin hydrochloride); and sulfonamide antibiotics (such as sulfacetamide, sulfadiazine, sulfadimidine, sulfisoxazole, and sulfadimidine). Sulfadoxin, sulfamethoxazole, sulfathiazole, sulfadiazine, sulfadiazine, sulfamethoxypyrimidine, sulfamethoxypyrimidine, sulfapyrazine, and terephthalyl, and their pharmaceutically acceptable salts, esters, or derivatives), β-lactam antibiotics (e.g., penicillin or its derivatives (e.g., penicillin G, penicillin V, benzylpenicillin, and phenoxymethylpenicillin), dicloxacillin, flucloxacillin, oxacillin, nafcillin, amoxicillin, ampicillin, ticarcillin, piperacillin, ritipenon, carbapenem (e.g., ertapenem, doripenon, imipenem, and meropenem),(and pharmaceutically acceptable salts, esters or derivatives thereof), cephalosporins (e.g., cefazolin, cefalexin, cefadroxil, cefepime, cefaclor, cefotetan, cefotaxime (cefoxitin), cefprozil, cefuroxime axetil, ceftriaxone, ceftazidime, cefoperazone, cefdinir, cefcarpine, cefodazole, cefotaxime, cefotaxime, cefpirome, cefpodoxime, cefbuprofen, ceftoranol, cefepime, cefuroxime, cefolomide, cefolomide, cefolomide, cefuroxime, cefepime, ceftiofur, cefquinolone and cefvitin, and pharmaceutically acceptable salts, esters or derivatives thereof), monoclonal antibodies. Aminocarbazin (e.g., aztreonam or its pharmaceutically acceptable salts, esters, or derivatives) and β-lactamase inhibitors (e.g., sulbactam, tazobactam, clavulanic acid, and avibactam or their pharmaceutically acceptable salts, esters, or derivatives) or cyclic peptide antibiotics (e.g., cyclosporine), therapeutic monoclonal antibodies or their therapeutic fragments (e.g., adalimumab, atorvaline, atezolizumab, bevacizumab, cannabinoids, caputuximab, cetrus, cetuximab, icovalizumab, efaizumab). lizumab), fontolizumab, girentuximab, golimumab, infliximab, labetuzumab, MABp1 (Xilonix™), natezumab, nimotuzumab, nivolumab, oregovomab, panitumumab Pembrolizumab, pemtumomab, pertuzumab, ramucirumab, ranibizumab, rituximab, lulizumab, tracalizumab, tocilizumab, trastuzumab, uslinta, vedozumab, vexizumab, vortumab, zalumumab, and zanomumab, and their active fragments, combinations, or conjugates), therapeutic fusion proteins (in some embodiments, recombinant fusion proteins,Examples include aflibercept (Regeneron), etanercept (Amgen), aflibercept (Astellas Pharma), abatacept (Bristol-Myers Squibb), lilonascept (Regeneron), romistastatin (Amgen), and beracide (Bristol-Myers Squibb), prostaglandin analogues (e.g., latanoprost, travoprost, tafluprost, unoprostone, netasudil, tatanoprostene bunod, netasudil, and bimatoprost, and their pharmaceutically acceptable salts, esters, and derivatives), growth factors (e.g., EGF, PDGF, TGF-β, IGF-1, VEGF, FGF-β, IGF-1) or their therapeutic or growth-promoting (especially skin growth-promoting) fragments, and neuropeptides (e.g., substance P (SEQ ID NO)). NO:389), α-adrenergic antagonists (e.g., brinzolamide, clonidine, and araclonidine, and their pharmaceutically acceptable salts, esters, or derivatives), β-adrenergic antagonists (e.g., timolol, propranolol, atenolol, levobenolol, carteolol, betalolol, and their pharmaceutically acceptable salts, esters, and derivatives, such as timolol maleate), cell surface receptor antagonists (e.g., lifitegrast or etanercept), carbonic anhydrase inhibitors (e.g., dzodamine, brinzolamide, acetazolamide, and acetazolamide, and their pharmaceutically acceptable salts, esters, or derivatives, such as timolol maleate), cell surface receptor antagonists (e.g., lifitegrast or etanercept), carbonic anhydrase inhibitors (e.g., dzodamine, brinzolamide, ... Pharmaceutically acceptable salts, esters, and derivatives, such as dzodamine hydrochloride, and their pharmaceutically acceptable salts, esters, and derivatives. For certain such therapeutic compounds, co-administration with the CMPs described herein (whether as a CMP-TC conjugate or simply mixed with one or more CMPs and one or more TCs or administered alone) can prevent, reduce, or mitigate one or more adverse side effects of the therapeutic compound. For example, therapeutic administration of certain fluoroquinolone antibiotics is known to cause damage to collagen and collagen-containing structures (e.g., tendons) in humans or animals treated with fluoroquinolones (see, for example, “FDA Drug Safety Communication: FDA updates warnings for oral and injectable fluoroquinolone antibiotics due to disabling side effects,” accessed November 6, 2017, at https: / / www.fda.gov / Drugs / DrugSafety / ucm511530.htm). Therefore, co-administration or co-administration with one or more CMPs described herein to humans or animals requiring treatment with fluoroquinolone drugs may prevent, reduce, or mitigate one or more adverse side effects of the therapeutic compound.CMP can enable patients to receive the therapeutic benefits of fluoroquinolone drugs while reducing, improving, or preventing collagen damage caused by such treatments, because it can target and repair areas of damaged collagen in the body.

[0484] Other suitable therapeutic compounds for use in the CMP-TC compounds, compositions, and conjugates of the present invention include other non-biological small molecule therapeutic compounds, including but not limited to alkylating agents, antitumor antibiotics, antimetabolites, hormones, alkaloids, angiogenesis inhibitors, GnRH agonists, tyrosine kinase inhibitors, etc. Examples of such non-biological small molecule therapeutic compounds suitably used according to the present invention include, but are not limited to, nitrosourea, lenalidomide, imatinib, benztroxetine valerate, bortezomib, abiraterone acetate, everolimus, taxol, docetaxel, paclitaxel, cabazitaxel, mitoxantrone, carboplatin, cisplatin, gemcitabine, doxorubicin, casrolide, flutamide, enzalutamide, abiraterone, sipuleucel-T, and ketoconazole. Other suitable non-biological small molecule therapeutic compounds that are beneficial for forming the CMP-TC conjugates of the present invention, and particularly for generating CMP-TC conjugates for treating certain cancers and preventing tumor metastasis, include inhibitors of lysyl oxidase (LOX), lysyl oxidase-like 1 (LOXL1), and lysyl oxidase-like 2 (LOXL2) enzymes. Such inhibitors have been suggested for potential therapeutic applications in the treatment and / or prevention of metastasis in certain cancers and solid tumors (see, for example, U.S. Patent Nos. 5,201,456, 5,120,764, 5,252,608, 8,461,303, 8,658,167, 8,680,246, 9,176,139, 9,255,086, and 9,289,447; see also Erler, JT, et al., Nature 440:1222-1226 (2006); Erler, JT, et al., Cancer Cell 15(1):35-44 (2009); Bondareva, A., et al., PLoS ONE 4(5):e5620 (2009); Granchi, C., et al., ChemMedChem 4(10):1590-1594). (2009); and Fang, M., et al., Tumor Biol. 35:2871-2882 (2014); the entire contents of which are incorporated herein by reference.

[0485] In a relevant aspect of the invention, CMP-TC conjugates comprising one or more LOX or LOX-like enzyme inhibitors are suitable for the treatment and / or prevention of certain fibrotic diseases and conditions in humans and animals mediated by oxidoreductases such as LOX and LOX-like enzymes (e.g., LOXL1 and LOXL2). Fibrous diseases and conditions suitably treated and / or prevented according to this aspect of the invention include, but are not limited to, pulmonary fibrosis, cirrhosis, myocardial fibrosis, surgical scars, systemic sclerosis, scleroderma, keloid formation, proliferative vitreoretinopathy, and other fibrotic diseases and conditions familiar to those skilled in the art. Particularly useful inhibitors of LOX and Lox-like proteins include β-aminopropionitrile and certain derivatives and prodrugs thereof (see, for example, U.S. Patent Nos. 5,201,456, 5,120,764, 5,252,608, 8,461,303, 8,680,246, 9,176,139 and 9,255,086, the entire disclosure of which is incorporated herein by reference), as well as antibodies (which may be polyclonal or preferably monoclonal) and fragments or portions thereof that bind to and inhibit the activity or function of LOX and LOX-like enzymes (see, for example, U.S. Patent No. 8,461,303, the entire disclosure of which is incorporated herein by reference).

[0486] In other embodiments, compounds or compositions comprising one or more CMPs and one or more antigens can be prepared, either as a mixture or co-formulated form of one or more CMPs with one or more antigens (and optionally with one or more pharmaceutically suitable carriers or excipients), or as other compounds or compositions in which one or more antigens are directly or indirectly linked or conjugated to one or more CMPs. According to some of these aspects, the antigen can be a complete antigen or an antigenic determinant or fragment thereof (e.g., a hapten) that, when presented to the immune system of a human or animal under appropriate physiological conditions, such as by administration of the compound, conjugate, or composition to a human or animal in the form of a vaccine or immunization, is capable of inducing an immune response in the human or animal. Compounds, conjugates, and compositions that can be used in such embodiments can be prepared by co-formulation or direct or indirect conjugation according to the methods described elsewhere herein for co-formulating and conjugating therapeutic compounds with CMPs. Suitable for such compounds, conjugates, and compositions, and therefore suitable for methods of their use, are antigens or portions thereof comprising any molecule or particle or portion thereof capable of inducing an immune response in humans or animals, including, but not limited to, antigens derived from or produced by bacteria (where the antigen may include the whole bacterium or a portion thereof, such as cell wall or cell membrane components, nuclear components, or bacterial toxins), viruses (where the antigen may include the whole viral particle or a portion thereof, such as a capsid component (e.g., a protein or lipid or a portion thereof), a nuclear component, or an enzyme encoded by or part of a viral particle), protozoa, fungi, plants (which may include plant irritants or allergens, such as pollen grains), animals (where the antigen or a portion thereof may be an allogeneic antigen or an autoantigen, or a portion thereof), etc.; examples of such antigens or portions thereof are readily familiar to those skilled in the art. Such compounds, compositions, or conjugates are suitable for methods of treating and / or preventing one or more conditions, diseases, and ailments in humans or animals, such as by inducing an immune response in humans or animals through the use of the compound, composition, or conjugate. In some such methods, diseases or conditions of animals or livestock are treated and / or prevented by administering one or more compounds, compositions, or conjugates of this aspect of the invention to humans or livestock (such as in the form of vaccines or immunizations). Such vaccines or immunizations are suitably formulated according to methods well known in the relevant art and administered in any manner that induces an immune response in humans or livestock against an antigen or a portion thereof, thereby treating and / or preventing diseases or conditions caused directly or indirectly by the antigen or a portion thereof.Such vaccines or immunizations can be administered to humans or animals via any suitable route, such as oral, parenteral (including subcutaneous, intradermal, transdermal, intrathecal, or intravenous), ocular administration (e.g., in the form of drops, gels, tablets, or by injection, as described elsewhere herein for CMP-TC administration to the eye), nasal administration, and other routes of administration familiar to those skilled in the art. In such embodiments, the compounds, conjugates, or compositions of the present invention are suitably administered to humans or animals until the humans or animals produce an immune response sufficient to treat and / or prevent the target disease or condition, and may be re-administered as needed to enhance the immune response and / or ensure sustained immunity against the target antigen or a portion thereof. Diseases and conditions appropriately treated by such methods of the present invention include any disease or condition involving or caused by any external factor acting on the cells, organs, organ systems, body structures, or body of humans and animals, including but not limited to infectious diseases, cancer, allergies and other excessive immune responses (e.g., graft-versus-host disease or host-versus-graft disease), Stevens-Johnson syndrome, mucosal pemphigoid, toxic epidermal necrolysis, Behçet's uveitis, shotgun retinochoroidal disease, juvenile idiopathic arthritis (JIA)-associated uveitis, multifocal choroiditis with panuveitis, necrotizing scleritis, creeping choroidal disease, sympathetic ophthalmia, Vogt-Koyanagi-Harada (VKH) disease, non-infectious panuveitis, etc.

[0487] Suitable diagnostic compounds that are linked or conjugated with CMP to produce the conjugates and compositions of the present invention include, but are not limited to, labeled probes, such as fluorescent dyes (e.g., quantum dots, indocyanine green, fluorescein, rhodamine, cyanine dyes, near-infrared fluorescent dyes, etc.); radioactive isotopes, nuclides for PET, nuclides for SPECT, and in particular, each radioactive isotope used for PET or SPECT is selected from... 11 C 13 N、 15 O、 18 F, 66 Ga、 67 Ga, 68Ga 60 Cu、 61 Cu、 62 Cu、 67 Cu、 64 Cu、 48 V, Tc-99m, 241 Am、 55 Co、 57 Co、 153 Gd, 111 In、 133 Ba、 82 Rb、 139 Ce, Te-123m137 Cs、 86 Y、 90 Y、 185 / 187 Re、 186 / 188 Re、 125 I. Their complexes and combinations thereof; and MRI contrast agents, CT contrast agents, and magnetic materials, particularly each of which is selected from the group consisting of gadolinium, Gd-DTPA, Gd-DTPA-BMA, Gd-HP-DO3A, iodine, iron, iron oxide, chromium, manganese, their complexes or chelates, and combinations thereof. According to such aspects of the invention, CMP and labeled probes are suitably physically or chemically bound directly to each other, for example by direct conjugation via coordination bonds, covalent bonds, hydrogen bonds, hydrophobic interactions, or physical adsorption, or by indirect conjugation using at least one linking means (such as those described herein and others known in the art). Methods for conjugating or linking diagnostic compounds to proteins (such as CMP) are known in the art (see, for example, U.S. Patent Application Publication No. US 2012 / 0195828 A1, the disclosure of which is incorporated herein by reference in its entirety).

[0488] Use of CMP and CMP-TC conjugates

[0489] Therefore, the present invention provides a method for preparing compositions that can be used to treat, prevent, diagnose, or improve diseases, conditions, or medical conditions in humans or animals. In another aspect, the present invention provides a method for treating, preventing, diagnosing, or improving diseases, conditions, or medical or physical conditions in humans or animals using the compositions of the present invention. Particularly preferred CMPs for these aspects of the invention include CMPs comprising, substantially composed of, or consisting of the following CMPs: (Pro-Pro-Gly)7 (SEQ ID NO: 1), (Flp-Pro-Gly)7 (SEQ ID NO: 4), (Pro-Flp-Gly)7 (SEQ ID NO: 5), (Flp-Hyp-Gly)7 (SEQ ID NO: 6), (Clp-Hyp-Gly)7 (SEQ ID NO: 9), (Hyp-Flp-Gly)7 (SEQ ID NO: 388), Gly3-(Pro-Hyp-Gly)6 (SEQ ID NO: 397), Gly3-(Pro-Flp-Gly)6 (SEQ ID NO: 398), Gly3-(Pro-Hyp-Gly)7 (SEQ ID NO: 399), Gly3-(Pro-Fl ... 400), Gly3-(Pro-Hyp-Gly)8 (SEQ ID NO: 401), Gly3-(Pro-Flp-Gly)8 (SEQ ID NO: 402), Gly3-(Pro-Hyp-Gly)9 (SEQ ID NO: 403), Gly3-(Pro-Flp-Gly)9 (SEQ ID NO: 404), (Pro-Hyp-Gly)6-Tyr (SEQ ID NO: 405), (Pro-Flp-Gly)6-Tyr (SEQ ID NO: 406), (Pro-Hyp-Gly)7-Tyr (SEQ ID NO: 407), (Pro-Flp-Gly)7-Tyr (SEQ ID NO: 408), (Pro-Hyp-Gly)8-Tyr (SEQ ID NO: 409), (Pro-Flp-Gly)8-Tyr (SEQ ID NO: 410), Cys-(Pro-Hyp-Gly)3 (SEQ ID NO: 411), Cys-(Pro-Flp-Gly)3 (SEQID NO: 412), Cys-(Pro-Hyp-Gly)5 (SEQ ID NO: 413), Cys-(Pro-Flp-Gly)5 (SEQ ID(SEQ ID NO: 414), Cys-(Pro-Hyp-Gly)7 (SEQ ID NO: 415) or Cys-(Pro-Flp-Gly)7 (SEQ ID NO: 416), and derivatives thereof containing one or more cysteine, methionine or lysine residues, such as those described elsewhere herein.

[0490] The CMP and CMP-TC conjugates of the present invention comprise solutions, gels, films, sheets, membranes, spheres, nanoparticles, and suspensions containing, substantially composed of, or composed of the CMP and / or CMP-TC conjugates of the present invention, and are suitable for use as medicaments for treating, preventing, or improving various diseases or conditions in humans or animals requiring treatment or prevention, or as compositions used as medicaments for treating, preventing, or improving various diseases or conditions in humans or animals requiring treatment or prevention. Other compositions provided in this aspect of the invention provide the use of CMP conjugated with one or more diagnostic compounds or molecules (such as one or more labeled probes) and then used as diagnostic reagents in various tests and assays, particularly in vivo or in situ tests and assays, to diagnose diseases, conditions, or physical conditions in humans or animals. Such pharmaceutical or diagnostic compositions, in addition to containing CMP, CMP-TC conjugates, or CMPs conjugated with one or more diagnostic compounds or molecules, may also contain one or more additional therapeutic compounds or pharmaceutically active ingredients (e.g., one or more antibiotics, one or more growth factors, growth factor-rich autologous plasma (PRGF), one or more cytokines, one or more antibody fragments, one or more non-biological small molecule therapeutic compounds, and their pharmaceutically active salts, esters, and derivatives, including those described herein and others known in the art). The compositions of the present invention may additionally or optionally contain one or more pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients suitable for the compositions and methods of the present invention include, for example, one or more solvents (which may include water, organic solvents, or inorganic solvents), one or more buffers, one or more polymers, one or more salts, one or more sugars, one or more sugar alcohols, one or more disintegrants, one or more aerosolizers or carriers, one or more desiccants, etc. Other pharmaceutically acceptable carriers or excipients suitable for the compositions of the present invention are readily familiar to those skilled in the art.

[0491] Not wishing to be bound by theory, it is believed that the CMP provided by this invention and used in the method of this invention is particularly suitable for repairing damaged collagen, especially damaged helical collagen, which is caused by or related to various diseases, conditions, structural abnormalities, physical conditions, and medical conditions in humans and animals. For example, when collagen is structurally damaged, it is usually hydrolyzed in one of the three helices that form triple-helix collagen, resulting in the unwinding of the triple helix and structural deformation. This destruction and digestion can also cause collagen to break down into many smaller fragments, which remain in the extracellular environment or enter the blood or lymphatic circulation system. These fragments are eventually either phagocytosed or bound by scavenger cells or bind to cell surface receptors on human or animal somatic cells. These receptors (which may include, for example, integrins, discoid domain receptors, glycoprotein VI, and leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1)) control cellular functions such as growth, differentiation, morphogenesis, tissue repair, adhesion, migration, homeostasis, immune function, and wound healing. They are frequently disrupted, or their functions or signaling systems are upregulated or downregulated by binding to these free collagen fragments. According to this theory, when CMPs encounter damaged collagen or fragments thereof, they dynamically anneal or bind to the broken collagen triple helix and repair its structure, thereby (among other things) leading to the restoration of normal function and signaling activity levels of the cellular receptors. Therefore, the overall result of applying CMPs in this manner to humans or animals suffering from diseases, conditions, structural abnormalities, or injuries involving or caused by damaged collagen is the release of an accelerated wound healing process, which, in some physiological contexts, includes the rapid growth, migration, and adhesion of epithelial cells, endothelial cells, or nerve cells on the now-repaired collagen matrix, leading to the restoration of normal or near-normal structure and function of such cells, as well as the tissues, organs, and organ systems containing such cells.

[0492] Diseases, conditions, physical conditions, and medical conditions appropriately treated, prevented, improved, or diagnosed using the compositions and methods of the present invention include, but are not limited to, eye diseases or conditions, neurological or nervous system diseases or conditions, and cardiovascular diseases or conditions. For such uses, suitable dosages of the compositions and conjugates of the present invention are those in which the concentration of the CMP component of the composition or conjugate is from about 10 ng / ml to about 500 µg / ml, from about 15 ng / ml to about 400 µg / ml, from about 20 ng / ml to about 300 µg / ml, from about 25 ng / ml to about 250 µg / ml, from about 30 ng / ml to about 200 µg / ml, from about 35 ng / ml to about 200 µg / ml, from about 40 ng / ml to about 200 µg / ml, from about 50 ng / ml to about 200 µg / ml, from about 75 ng / ml to about 200 µg / ml, and from about 100 ng / ml to about 200 µg / ml. In some such embodiments, the conjugate or composition is suitable for application to the treated tissue, organ, or organ system at a dose equivalent to a CMP concentration of about 25 μg / ml to about 500 μg / ml (e.g., about 25 μg / ml, about 30 μg / ml, about 35 μg / ml, about 40 μg / ml, about 45 μg / ml, about 50 μg / ml, about 75 μg / ml, about 100 μg / ml, about 125 μg / ml, about 150 μg / ml, about 175 μg / ml, about 200 μg / ml, about 225 μg / ml, about 250 μg / ml, about 300 μg / ml, about 350 μg / ml, about 400 μg / ml, about 450 μg / ml, or about 500 μg / ml). In certain implementations of this type, concentrations of CMP equivalent to about 25 µg / ml, 50 µg / ml, 75 µg / ml, or 100 µg / ml are used. Appropriate doses of CMP for human administration (per kilogram of body weight) are about 10 ng / kg to about 1000 ng / kg, about 15 ng / kg to about 650 ng / kg, about 20 ng / kg to about 600 ng / kg, about 25 ng / kg to about 550 ng / kg, about 30 ng / kg to about 500 ng / kg, about 20 ng / kg to about 350 ng / kg, and about 25 ng / kg to about 300 ng / kg, about 10 ng / kg to about 250 ng / kg, about 10 ng / kg to about 200 ng / kg, about 10 ng / kg to about 150 ng / kg, or about 10 ng / kg to about 100 ng / kg. When treating specific cardiovascular, cerebrovascular, or nervous system diseases, conditions, or illnesses in humans or animals, clinicians may also determine the appropriate specific concentration to be administered within these ranges by titration, using methods familiar to licensed physicians and general technicians in the relevant pharmaceutical and medical fields.Those skilled in the art can readily determine additional concentrations, dosages, and amounts of the conjugates or compositions of the present invention suitable for such methods, based on the information contained herein and available in the art, without resorting to excessive experimentation.

[0493] The compositions and methods of the present invention can be used to treat, prevent, improve, or diagnose ocular diseases or conditions, including but not limited to those involving the cornea and / or sclera, i.e., corneal and scleral diseases, conditions, and ailments. Without wishing to be bound by theory, it is believed that the compositions of the present invention, particularly one or more CMPs contained in such compositions, can repair disordered, disordered, or digested collagen found in the extracellular matrix and / or matrix of the cornea and sclera, thereby repairing the collagen matrix / matrix and restoring the homeostatic structure and function (including hardening or strengthening) of the cornea and / or sclera, thereby treating or improving such diseases and conditions and their signs / symptoms. In other aspects, the compositions of the present invention, particularly one or more CMPs contained in such compositions, prevent the disintegration and digestion of collagen in the extracellular matrix and / or matrix of the cornea and sclera upon damage or injury, thus preserving the collagen matrix / matrix and therefore the homeostatic structure and function of the cornea and / or sclera, thereby preventing such diseases and conditions and their signs / symptoms. Examples of corneal and scleral diseases, conditions, and ailments appropriately treated, prevented, improved, or diagnosed using the compositions and methods of the present invention include, but are not limited to, myopia, presbyopia, hyperopia, keratitis, episcleritis, scleritis, corneal ulcer, sequelae of corneal ulcer, corneal ectasia, acquired corneal morphology, keratoconus, corneal astigmatism, spherical keratosis, posterior corneal apnea, corneal bulging, posterior elastic bulging, clear marginal keratosis, Terrien's marginal dystrophy, Mooren's ulcers, central corneal ulcer, marginal corneal ulcer, staph marginal ulceration, and Salzman's nodular dystrophy. This invention relates to various eye conditions, including age-related peripheral corneal atrophy, geographic ulcers, discoid stromal keratitis, herpetic ulcers, keratomalacia, post-penetrating keratoplasty, surgical wounds, anterior membrane dystrophy, stromal dystrophy, ocular mucosal pemphigoid, necrotizing scleritis, scleromalacia, coloboma, scleral buckling-induced scleromalacia, congenital hereditary stromal dystrophy, congenital anterior staphyloma, scleralized keratoplasty, traumatic desemmel membrane rupture, corneal keloids, scleral bulging, scleral staphyloma, deep scleritis, necrotizing scleritis, penetrating scleromalacia, scleral hyalinization, paralimbal scleromalacia, graft-versus-host disease, and choroidal agenesis. Specifically, the compositions and methods of this invention are suitable for the treatment, improvement, prevention, and / or diagnosis of myopia, presbyopia, and keratoconus.Based on information readily available in the literature, it will be apparent to those skilled in the art that other corneal and scleral diseases, conditions, and ailments can be suitably prevented, treated, improved, or diagnosed using the compositions and methods of the present invention.

[0494] Other ocular diseases or conditions, including anterior segment diseases and conditions, that can be treated, prevented, improved, or diagnosed using the compositions and methods of the present invention, include but are not limited to glaucoma, cataracts, vitreous adhesions or floaters, macular degeneration, dry eye syndrome (also known as dry eye disease), keratitis, non-infectious corneal ulcers, non-infectious corneal ablation, infectious corneal ulcers, infectious corneal ablation, conjunctivitis, Stevens-Johnson syndrome, iritis, uveitis, vitreitis, and Behçet's uvea. Uveitis, shotgun retinal choroidal disease, juvenile idiopathic arthritis (JIA)-associated uveitis, multifocal choroiditis with panuveitis, necrotizing scleritis, creeping choroidal disease, sympathetic ophthalmia, Vogt-Koyanagi-Harada (VKH) disease, non-infectious panuveitis, ectasia, corneal laceration, corneal erosion, corneal abrasion, acute or chronic corneal pain (especially pain caused by corneal nerve damage or injury or denervation; see, for example, Rosenthal, P. and Borsook, D., Br J Ophthalmol. 2016;100(1):128-134; Theophanous, C., et al., Optom. Vis. Sci. 2015;92(9):e233-240; Belmonte, C., et al., Ocul. Surf.2004;2(4):248-253; Belmonte, C., et al., Exp. Eye Res. 2004;78(3):513-525; Belmonte, C., et al., Curr. Ophthalmol Rep. 2015;3(2):111-121; Baratta, RO, et al., Front. Pharmacol. 12:70563 (August 16, 2021), doi:10.3389 / fphar.2021.705623;Baratta, RO et al., Surv. Ophthalmol. 67:60-67 (2022); Ribeiro, M et al., Int. J.Mol. Sci. 23:2911 (2022); McGrady, NR et al., Front. Pharmacol. 12:8764709 (November 2, 2021), doi:10.3389 / fphar.2021.764709), including but not limited to periorbital pain, extraocular pain, and postherpetic neuralgia, as well as postoperative ocular pain caused by ophthalmic surgery. Such postoperative ocular pain caused by ophthalmic surgery can be, for example, pain following cataract surgery or glaucoma surgery, especially those that cause or are themselves required to treat a postoperative ocular condition with medication.

[0495] Other ocular diseases or conditions that can be treated, prevented, improved, or diagnosed using the compositions and methods of the present invention include, but are not limited to, diseases and conditions of the posterior segment of the eye, particularly those involving the retina, including, but not limited to, macular degeneration (wet, dry, and age-related), retinitis pigmentosa, retinal tears or detachments, retinopathy (e.g., diabetic retinopathy), arterial or venous occlusions (e.g., BRAO (branch retinal artery occlusion), CRAO (central retinal artery occlusion), BRVO (branch retinal vein occlusion), and CRVO (central retinal vein occlusion), optic neuritis, and optic neuropathy (including, for example, AION). (Anterior ischemic optic neuropathy) and traumatic optic neuropathy), optic atrophy (e.g., glaucomatous optic atrophy), one or more diseases, conditions, or illnesses affecting the eye or surrounding areas, including peripheral eye diseases, conditions, or illnesses, and extraocular diseases, conditions, or illnesses, such as cranial nerve palsy including but not limited to cranial nerve III palsy, cranial nerve IV palsy, cranial nerve V palsy (e.g., trigeminal neuralgia and postherpetic neuralgia), cranial nerve VI palsy and cranial nerve VII palsy (e.g., Bell's palsy), and other retinal and posterior segment-related conditions and illnesses involving the retinal epithelium (particularly the retinal pigment epithelium, retinal vessels and / or retina, cranial nerves or optic nerves). Other ocular conditions that can be advantageously treated, improved and / or prevented using the compositions and methods of the present invention include, but are not limited to, refractive eye diseases, including myopia, presbyopia and amblyopia. For example, myopia is known to be associated with axial elongation, which may involve in part the destruction of collagen in the sclera (see, for example, Guo, P. et al., Trans. Vis. Sci. Tech. 9(9):45). (2020); Zhao, F. et al., Am. J. Pathol. 188:1754-1767 (2018)), and can be improved by scleral reinforcement through cross-linking of scleral connective tissue or other manipulations (see, for example, Backhouse, S. et al., Ann. Eye Sci. 3:5 (2018); Grytz, R. et al., Curr. Opin. Biomed. Eng. 15:40-50 (2020); Garcia, MB et al., Invest. Ophthalmol. Vis. Sci. 58:1875-1886 (2017)).

[0496] According to this aspect of the invention, a method of treating or preventing eye diseases, conditions, or wounds in humans or animals that are susceptible to or prone to them comprises applying the compositions described herein, particularly CMP or CMP-TC conjugates and / or compositions containing such conjugates, to the eyes of the human or animal. Not wishing to be bound by theory, the inventors hypothesize that in the region of an eye disease or condition, the degree of damage to type I collagen is sufficiently great that the CMP will specifically target the site of the eye disease or condition and insert into the collagen structure, for example by inserting into one or more damaged helices of helical collagen, thereby directly reforming functional collagen helices or matrix, or, in the case of a CMP conjugated with a therapeutic compound, delivering the therapeutic compound to the site where it must function to treat, prevent, or improve the eye disease or condition. In certain such anterior segment eye diseases or conditions, such as acute or chronic corneal pain (including but not limited to periorbital pain, extraocular pain, and postherpetic neuralgia), the denervated cornea has poor healing capacity, and therefore topical therapies that can influence nerve regeneration will be popular in this field. Pain, whether acute or chronic, is caused by damaged corneal nerves (see, for example, Rosenthal, P. and Borsook, D., Br J Ophthalmol. 2016;100(1):128-134; Theophanous, C., et al., Optom. Vis. Sci. 2015;92(9):e233-240; Belmonte, C., et al., Ocul. Surf. 2004;2(4):248-253; Belmonte, C., et al., Exp. EyeRes. 2004;78(3):513-525; Belmonte, C., et al., Curr. Ophthalmol Rep. 2015;3(2):111-121), therefore, treatments beneficial to neurological health are clinically valuable for such patients. Based on the findings described herein regarding the behavior of dorsal root ganglion cells exposed to certain CMPs of the present invention following damage to the collagen support layer (see Example 1 below), it can be anticipated that any cranial nerve will behave in a similar manner. Therefore, corneal nerves, branches of the trigeminal nerve, will benefit from therapies comprising topical application of one or more of the CMPs or CMP-TC conjugates described herein to the cornea. With nerve repair and regeneration, corneal recovery and pain relief will follow, thereby improving acute or chronic corneal pain.

[0497] The conjugate or composition is suitably applied to the eye at a concentration, dose, or amount (such as those described above) sufficient to treat or prevent eye diseases, conditions, or wounds, and the eye condition of the human or animal is then monitored over time to observe improvement in the disease state or physical condition. If necessary, the conjugate or composition of the present invention is periodically reapplied to the eye according to the dosing and treatment schedule and protocols described herein, as well as other protocols familiar to those skilled in the art, until the eye disease, condition, or wound is cured, prevented, or improved. In such embodiments, the conjugate or composition of the present invention for treating anterior segment diseases and conditions can be suitably applied to the surface of the eye via the conjunctiva or subconjunctiva, particularly by dropwise application of the conjugate or composition to the surface of the eye or the subconjunctival fornix. In other embodiments, the conjugate or composition of the present invention for treating anterior segment diseases and conditions can be suitably applied to the anterior portion of the eye (including the sclera and vitreous body) by drop (depending on the ability of the CMP-containing formulation to migrate across the surface epithelium) or by injection, such as intravitreal injection, in accordance with methods well known to those skilled in the art of medicine and pharmacy. In other embodiments relating to the treatment, prevention, cure, or diagnosis of posterior segment diseases and conditions, the conjugates and compositions of the present invention may be applied to the posterior segment of the eye, such as at or near the retina, via mechanical introduction (e.g., injection using a needle or other suitable device), or by applying the conjugates or compositions to the ocular surface in the form of drops, wherein the conjugates or compositions (or components thereof, such as CMP or CMP-TC conjugates) are transported or migrated to the posterior segment of the eye (e.g., at or near the retina). Application of the conjugates or compositions to the eye may be accomplished by any well-known means, including applying the conjugates or compositions to the eye in the form of a solution, gel, or suspension containing the composition or conjugate as one or more drops or aliquots; by injection; in the form of a solid material, such as a sheet or film (e.g., those described herein), implanted into the ocular structures; in the form of a mesh or patch; or by attaching or encapsulating the conjugates or compositions in one or more gels, spheres, or nanoparticles and then delivering them to the ocular structures. Other suitable methods of applying conjugates or compositions to the eye to achieve the therapeutic and diagnostic methods of the present invention will be apparent to those skilled in the art.

[0498] The compositions and methods of the present invention can be used to treat, prevent, improve, reverse, or diagnose diseases or conditions of the nervous system (including the central nervous system (“CNS”) and the peripheral nervous system (“PNS”)), including but not limited to damage to one or more nerves or nerve processes (including axons, dendrites, and neurons or neuronal bodies, ganglia, nerve bundles, etc.); neurodegenerative diseases, injuries, or conditions, including diseases, injuries, or conditions that may involve the cerebrovascular system (in many different physiological or disease conditions, such as multiple sclerosis, amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, traumatic encephalopathy, non-Alzheimer's dementia (including but not limited to Lewy body dementia), encephalitis, meningitis, etc.); diseases or injuries involving the peripheral nerves (such as peripheral neuropathy, such as diabetic peripheral neuropathy, trophic neuropathy, and alcohol-induced neuropathy, or damage caused by the severance or compression of one or more peripheral nerves). Injury); or certain neuro-ocular diseases and conditions, including those involving or affecting corneal nerves, retinal nerves, and optic nerves, including but not limited to glaucoma, macular degeneration (wet and / or dry, possibly age-related or unrelated), neurotrophic keratitis, retinopathy (which may include diabetic retinopathy, ischemic retinopathy, proliferative retinopathy, geographic atrophy, and other hereditary retinopathy and hereditary retinal diseases or conditions known in the art); damage or inflammation of one or more corneal nerves (which may result from eye injury or inflammation caused by external disease or trauma / wound, including transverse, crush, or torsional damage to nerves or nerve processes); corneal pain (which may be acute or chronic and may be caused by damage or injury to corneal nerves or corneal denervation, such as periocular pain, extraocular pain, and postherpetic neuralgia); encephalopathy (e.g., traumatic encephalopathy, such as concussion, encephalitis, meningitis), etc. In some such embodiments, the compositions and methods of the present invention can be used to induce nerve repair or regeneration (e.g., through nerve regeneration), particularly in cranial nerves, including but not limited to the optic nerve, retinal nerves, auditory nerves, or spinal nerves. In other such embodiments, the compositions and methods of the present invention can be used to protect certain nerves from degeneration or from further or ongoing degeneration (i.e., to provide neuroprotective function), which can be used, for example, to prevent, reduce, or slow the progression of peripheral nerve degeneration to prevent and / or treat diabetic peripheral neuropathy, trophic neuropathy, and alcohol-induced peripheral neuropathy, as well as the degenerative progression of corneal nerves, optic nerves, and / or retinal nerves to prevent and / or treat corneal pain (e.g., acute or chronic corneal pain, including but not limited to periocular pain, extraocular pain, and postherpetic neuralgia), glaucoma, hereditary retinal diseases or conditions, and genetically based retinal diseases (e.g., diabetic retinopathy).Based on the guidance provided herein and taking into account the information readily available in the relevant fields, those skilled in the art will recognize other beneficial uses of the compositions and methods of the present invention in the treatment, prevention, improvement, or diagnosis of neurological and nervous system diseases or conditions.

[0499] According to this aspect of the invention, a method of treating or preventing neurological or nervous system diseases or conditions in humans or animals suffering from or susceptible to such diseases or conditions comprises applying the compositions described herein (particularly CMP and CMP-TC conjugates, and compositions comprising such CMP and / or conjugates) to one or more tissues, organs, or organ systems of a human or animal or their vicinity at a site near or in connection with the neurological or nervous system disease or condition, or the site of the neurological or nervous system lesion causing the neurological or nervous system disease or condition. Not wishing to be bound by theory, the inventors hypothesize that in certain areas of neurological or nervous system diseases and conditions, collagen (possibly in other components of the local extracellular matrix) is sufficiently disrupted such that CMP will specifically target the site of the neurological or nervous system disease or condition and insert into the collagen structure, thereby directly inducing nerve regeneration and / or neuroprotection through the reformation of the functional collagen matrix, or, in the case where the CMP carries a therapeutic compound, delivering the therapeutic compound to the site where it must function to treat, prevent, or improve the neurological or nervous system disease or condition. According to this aspect of the invention, the conjugate or composition is suitably applied to a tissue, organ, or organ system, its vicinity, or within it at a dose, concentration, and amount (such as those described above) sufficient to treat, prevent, or improve a neurological or nervous system disease or condition, and then the progression, relief, or stagnation of the neurological or nervous system disease or condition in a human or animal is monitored over time to observe improvement in the disease or condition status. Those skilled in the art can readily determine additional concentrations and amounts of the conjugate or composition of the invention suitable for such methods based on the information contained herein and available in the art, without resorting to excessive experimentation. If necessary, the conjugate or composition of the invention is periodically reapplied to one or more tissues, organs, or organ systems, its vicinity, or on it, according to the dosing and treatment schedules and protocols described herein and protocols familiar to those skilled in the art, until the neurological or nervous system disease or condition is cured, prevented, or improved. In such embodiments, the conjugate or composition of the invention is suitably applied to a tissue, organ, or organ system, its vicinity, or within it, either parenterally or topically. Parenteral administration is achieved by any route of administration known in the art for the treatment of a tissue, organ, or organ system, such as by a route selected from the group consisting of: subcutaneous injection, intradermal injection, intramuscular injection, intracranial injection, intraspinal injection, or injection into any tissue, organ, or organ system with a neurological or nervous system disease or condition; intravenous infusion; intra-arterial infusion; endoscopic application; transdermal diffusion; implantation of drug-eluting sheets, films, gels, or putties; sublingual; oral; or rectal.In some such methods, the composition is appropriately parenterally applied to humans or animals in the form of an injectable solution or paste, pill, capsule, solution, suspension, or powder for inhalation or ingestion, or in the form of a mesh, film, sheet, gel, sphere, nanoparticle, paste, putty, or patch implanted near, on, or within a site of nerve or nervous system disease or symptom. In some such embodiments, one or more compounds, compositions, or conjugates of the invention may be coated on or within a mesh or "cannula" material such that the mesh or cannula material is impregnated with one or more compounds, compositions, or conjugates of the invention, and then the mesh or cannula is applied to an injured (e.g., transverse) or damaged nerve, neurite, or nerve bundle. In some such aspects of the invention, the compositions and methods of the invention are used to improve the integrity of nerves and blood vessels (including the cerebrovascular system in the brain) associated with the PNS and CNS, and to reverse their degradation, thereby treating, preventing, and / or reversing certain CNS and PNS diseases and symptom, such as those described herein.

[0500] In other embodiments, the application of the conjugate or composition to, near, or within a tissue, organ, or organ system can be achieved by any well-known means, including in the form of solutions, ointments, creams, patches, films, gels, pastes, spheres, nanoparticles, putty, creams, topical solutions, and drug-eluting sheets. For example, the conjugate or composition can be applied to or introduced into, a tissue, organ, or organ system, or its vicinity, as a drop or more of a solution or suspension containing the composition or conjugate (e.g., for the posterior part of the eye, as a topical ophthalmic drop); by injection; as a coating on a solid material implanted within, near, or on a tissue, organ, or organ system; as a mesh or patch; or by attaching or encapsulating the conjugate or composition in one or more nanoparticles, and then delivering the nanoparticles to, near, or on the tissue, organ, or organ system. Other suitable methods of applying the conjugate or composition to, on, near, or within a tissue, organ, or organ system to achieve the therapeutic and diagnostic methods of the present invention will be apparent to those skilled in the art.

[0501] The compositions and methods of the present invention can be used to treat, prevent, improve, or diagnose cardiovascular diseases or conditions, including but not limited to myocardial infarction, heart failure, valvular heart disease, atherosclerosis (including but not limited to diabetic atherosclerosis), cardiomyopathy, arrhythmia, congenital heart disease, coronary artery disease, pericardial disease, vascular occlusive diseases (e.g., affecting the carotid artery, aorta, cerebral vascular system (especially the cerebrovascular system), renal artery, femoral artery, pulmonary artery, and other large and small blood vessels, which may be arteries, arterioles, veins, venules, etc.), cerebrovascular diseases (including diabetic cerebrovascular diseases and conditions or traumatic injuries, such as diseases and conditions characterized by blockage, trauma, or leakage of the cerebral vascular system, such as stroke, aneurysm, cerebral hemorrhage, cerebral microbleeds, chronic traumatic encephalopathy, etc.), hemorrhagic diseases (e.g., arterial, venous, or capillary leakage, including microbleeds in tissues and organs such as the brain, skin, heart, and other tissues, organs, and organ systems), Marfan syndrome, etc.

[0502] According to this aspect of the invention, a method of treating or preventing cardiovascular disease or condition in humans or animals suffering from or susceptible to such disease or condition comprises applying the compositions described herein (particularly CMP and / or CMP-TC conjugates and compositions comprising such CMP and / or conjugates) to the vascular system of humans or animals suffering from or susceptible to such disease or condition. Not wishing to be bound by theory, the inventors hypothesize that in certain areas of cardiovascular disease or condition, type I collagen is sufficiently disrupted such that CMP introduced into the subject's vascular system will specifically target the site of cardiovascular disease or condition and embed itself within the collagen structure, thereby directly reforming the functional collagen matrix, or, in the case of CMP conjugated with a therapeutic compound, thereby delivering the CMP and / or therapeutic compound to the site where it must function to treat, prevent, or improve the cardiovascular disease or condition. According to this aspect of the invention, the conjugate or composition is suitably administered into the vascular system at a dose, concentration, and amount (such as those described above) sufficient to treat, prevent, or improve cardiovascular disease or condition, and then the progression, relief, or stagnation of the cardiovascular disease or condition in a human or animal is monitored over time to observe improvement in the disease or condition status. If necessary, the conjugate or composition of the invention is periodically re-administered into the vascular system according to the dosing and treatment schedule and protocols described herein, as well as other protocols familiar to those skilled in the art, until the cardiovascular disease, condition, or wound is cured, prevented, or improved. In such embodiments, the conjugate or composition of the invention is suitable for parenteral or topical administration to or within the heart, pericardium, blood vessels, or other relevant components of the vascular system. Parenteral administration can be achieved via any route of administration of therapy to the vascular system known in the art, such as by routes selected from the group consisting of: oral ingestion, subcutaneous injection, intradermal injection, transdermal administration or diffusion (e.g., via ointment, patch, etc.), intramuscular injection, intravenous infusion, intra-arterial infusion, via catheter insertion, embolization, inhalation, intrathecal administration, implantation of drug-eluting sheets or films, sublingual or rectal administration. In such methods, the composition is suitable for parenteral administration to humans or animals in the form of pills, capsules, solutions, suspensions or powders for ingestion by humans or animals, or in the form of meshes, sheets, films, gels, putties, spheres, nanoparticles or patches implanted in or near the heart, pericardium, blood vessels or other relevant components of the vascular system involved in cardiovascular disease or condition.

[0503] In other embodiments, the application of the conjugate or composition to or within the vascular system can be achieved by any well-known means, including solutions, ointments, creams, patches, films, gels, spheres, nanoparticles, creams, topical solutions, and drug-eluting sheets. For example, the conjugate or composition can be applied to or introduced into the heart, pericardium, blood vessels, or other relevant components of the vascular system as one or more drops of a solution or suspension containing the composition or conjugate; by injection; implanted as a coating on a solid material; in the form of a mesh or patch; or by attaching or encapsulating the conjugate or composition into one or more nanoparticles, followed by delivery of the nanoparticles to the heart, pericardium, blood vessels, or other relevant components of the vascular system. Other suitable methods for applying the conjugate or composition to or within the vascular system to achieve the therapeutic and diagnostic methods of the present invention will be apparent to those skilled in the art.

[0504] In relevant embodiments, the present invention provides devices, particularly medical devices, suitable for treating or preventing diseases, conditions, or medical conditions in humans or animals that are susceptible to or have such diseases, symptoms, or medical conditions. Such devices suitably comprise at least one composition of the present invention, either as a coating on the device or embedded within the device, such that it is released or washed out of the device upon implantation into a human or animal. Suitable such devices include, but are not limited to, artificial joints, braces, catheters, sutures, bone screws, bone plates, prostheses (e.g., artificial limbs, body structures, organs, etc.), absorbable or non-absorbable meshes, absorbable or non-absorbable patches, drug-release sheets, brain stimulators (e.g., deep brain stimulators), gastric stimulators, cochlear implants, defibrillators, pacemakers, insulin pumps, internal infusion pumps, etc. Other suitable devices useful according to this aspect of the invention will be apparent to those skilled in the art.

[0505] The apparatus provided in this aspect of the invention can be used to treat, prevent, improve, or diagnose diseases, conditions, or medical conditions in humans or animals that are susceptible to or prone to such diseases, conditions, or medical conditions. According to the method of this aspect, one or more medical devices of the invention are implanted into a human or animal, and the medical condition of the human or animal is monitored until the disease, condition, or medical condition of the human or animal is cured, improved, or prevented. Suitable diseases, conditions, and medical conditions that can be cured, treated, improved, or prevented using the devices and methods of the present invention include cancer (e.g., cancers described elsewhere herein), and diseases or conditions affecting organ systems in humans or animals, including the exodermal system (particularly skin diseases or conditions, such as those described in detail herein), the muscular system, the skeletal system (particularly diseases or conditions of bones, joints, cartilage, tendons, or ligaments, such as those described in detail herein), the nervous system (particularly the nervous system of the brain or eye (e.g., anterior segment eye diseases and conditions, including but not limited to diseases and conditions involving corneal nerves (e.g., corneal pain (which can be acute or chronic), including but not limited to diseases caused by damage or loss of corneal nerve innervation, such as periorbital pain, extraocular pain, and postherpetic neuralgia), glaucoma, cataracts, vitreous adhesions or floaters, macular degeneration, dry eye, keratitis, non-infectious corneal ulcers, non-infectious corneal ablation, infectious corneal ulcers, infectious corneal ablation, conjunctival... Inflammation, Stevens-Johnson syndrome, scleritis, episcleritis, iritis, uveitis, vitreitis, Behçet's uveitis, shotgun retinochoroidal disease, juvenile idiopathic arthritis (JIA)-associated uveitis, multifocal choroiditis with panuveitis, necrotizing scleritis, creeping choroidal disease, sympathetic ophthalmia, Vogt-Koyanagi-Harada (VKH) disease, non-infectious panuveitis, corneal ectasia, keratoconus, corneal laceration, corneal erosion, corneal abrasion, and other eye conditions. Postoperative eye conditions following procedures such as cataract surgery or glaucoma surgery requiring medication, or posterior segment eye diseases involving the retina, retinal epithelium (especially retinal pigment epithelium), retinal vessels, retinal nerves, or optic nerve, including but not limited to macular degeneration (wet, dry, and age-related), retinitis pigmentosa, retinal tears and detachments, retinopathy (e.g., diabetic retinopathy), retinal artery or vein occlusion (e.g., BRAO, CRAO, BRVO, and CRVO), optic neuritis, optic neuropathy (including, for example, AION, traumatic optic neuropathy, and visual atrophy (e.g., glaucomatous optic atrophy)), and other neuropathy affecting the eye or surrounding areas.This includes diseases, conditions, and medical conditions affecting the periocular region, as well as diseases, conditions, and medical conditions outside the eye, such as cranial nerve palsies, including but not limited to cranial nerve III palsy, cranial nerve IV palsy, cranial nerve V palsy (e.g., trigeminal neuralgia and postherpetic neuralgia), cranial nerve VI palsy, and cranial nerve VII palsy (e.g., Bell's palsy), circulatory system, lymphatic system, respiratory system (including diseases or conditions affecting the epiglottis, trachea, bronchi, bronchioles, or lungs in humans or animals, especially those detailed in this document), endocrine system, urinary / excretory system (including diseases or conditions affecting the kidneys, ureters, bladder, upper bladder, etc.). The urinary tract (i.e., the renal pelvis), the ureters or urethra of humans or animals, especially those diseases and conditions detailed herein), the reproductive system (including diseases and conditions affecting the testes, prostate, penis, vagina, cervix, uterus, fallopian tubes, or ovaries of said humans or animals, especially those diseases and conditions detailed herein), the digestive system (including diseases or conditions affecting the esophagus, stomach, small intestine, colon, or rectum of said humans or animals, especially those diseases and conditions detailed herein), and the nervous system (including the peripheral nervous system and the central nervous system, especially those nervous system diseases, ailments, and injuries detailed herein). Suitable methods for implanting one or more devices provided in this aspect of the invention into humans or animals to achieve the treatment, prevention, improvement, or diagnosis of diseases, conditions, or medical or physical conditions in humans or animals are well known to those skilled in the art in the relevant medical and surgical fields.

[0506] The concentrations of CMPs or CMP-TC conjugates that can be used to treat, prevent, improve, or diagnose one or more diseases or conditions according to the methods of the present invention will be apparent to those skilled in the pharmaceutical and medical fields, especially given the guidance provided herein. For unconjugated CMPs, the appropriate amount or concentration of CMP to be administered to a subject (particularly a human or animal), the appropriate amount or concentration of CMP to be used, includes those described above. Based on the guidance provided herein, those skilled in the medical, pharmaceutical, and / or pharmacological fields can determine the appropriate amount of the conjugates and compositions of the present invention to be used per kilogram (kg) of human or animal body weight. For conjugated CMP-TCs, the same amount or concentration of CMP described herein, whether concentration (e.g., ng / ml or µg / ml) or amount (e.g., mg / kg body weight), is suitable for administration to a subject, and the amount of active pharmaceutical ingredient or biological agent is calculated during the conjugation process to deliver a therapeutically effective amount of the desired active pharmaceutical ingredient or biological agent, specifically depending on the disease or condition to be treated, prevented, improved, or diagnosed in the human or animal. The appropriate amount or concentration of the active pharmaceutical ingredient or biological agent used in this aspect of the invention is familiar to those skilled in the art and can be readily determined based on the information contained herein and other information available in the relevant fields.

[0507] It will be apparent to those skilled in the art that other suitable modifications and adjustments can be made to the methods and applications described herein without departing from the scope of the invention or any of its embodiments. The invention has now been described in detail, and will be more clearly understood by referring to the following examples, which are included herein for illustrative purposes only and are not intended to limit the invention.

[0508] Example

[0509] Example 1: The role of CMP and CMP-TC conjugates in the treatment of neurological disorders

[0510] To examine the potential therapeutic effects of the CMP of the present invention in treating neurological disorders, studies were designed to test the ability of certain CMP conjugates to stimulate nerve growth in various in vitro environments.

[0511] In the initial studies of this kind, dorsal root ganglion (DRG) neurons, which are components of the peripheral nervous system, were isolated from day 19 rat embryos (Berta, T et al., Expert Opin. Ther. Targets 21:695-703 (2017)) and plated onto tissue culture plates coated with intact collagen or collagen digested with MMP-1. The plates were then treated with a medium (PBS) or with one of the different CMPs of the present invention: CMP 05A (SEQ ID NO:3), CMP 09C (SEQ ID NO:5), CMP 10A (SEQ ID NO:473), or CMP 13A (SEQ ID NO:6), each at 100 µM, 100 µL per well, and incubated at 37°C for five hours. The plates were then rinsed with culture medium, and DRG neurons were evenly seeded onto plates with one of three surface coating conditions: (1) intact collagen; (2) MMP1-digested collagen; and (3) MMP1-digested collagen + CMP. The plates were then incubated at 37°C for 24 or 48 hours, and morphology and neurite growth were examined by inverted bright-field microscopy. Representative results are presented in... Figure 1 As shown in the image.

[0512] After 24 hours, a phase-contrast micrograph of the collapsed Z-stacked montage of the DRG explants on intact collagen showed moderate neurite growth, which significantly increased by 48 hours. Figure 1 A, E). Lamination on damaged collagen reduces the degree of growth ( Figure 1 B, F), while explants plated on damaged collagen treated with two different CMPs (CMP 09C or CMP 10A) showed improved growth ( Figure 1 C, G and Figure 1 D, H).

[0513] To accurately measure neurite growth, multifocal high-magnification phase-contrast microscopy was used to visualize the explant in vivo to optimize the longest neurite. Figure 2 A (white arrow) and identification and measurement of the size of the DRG explant growth region. This method is superior to post-fixation measurement of neurites visualized after fixation using antibodies against β3 tubulin. Figure 2 B).

[0514] These trends were quantified by measuring the longest neurite extending from each explant and the area of ​​the neurite-covered region extending from and surrounding the explant, both normalized relative to values ​​measured under intact collagen conditions. The longest neurite of the medium-treated explants plated on MMP1-digested collagen was 15% shorter than that of intact collagen (p=0.014). Figure 3 A). In contrast, two of the four CMPs tested under these conditions induced neurite growth that was even 29% (CMP 10A) and 21% (CMP 09C) larger than that on intact collagen (p≤0.036). Similarly, explants treated with the medium showed a 30% smaller growth area compared to explants on intact collagen (p<0.001). Figure 3 B). Similarly, compared with the mediator, both CMPs significantly improved growth (p≤0.03), and one of them (CMP10A) also induced larger neurite growth.

[0515] These results are reminiscent of some of the results previously proposed by the inventors of this invention (see U.S. Patent Publication No. US 2020 / 0353056 A1, the disclosure of which is incorporated herein by reference in its entirety). In those prior studies (such as...) Figure 4 As shown, fluorescently labeled CMPs injected intravitreally or instilled into the mouse eye exhibited highly directional binding at and around ganglion neurons in the retinal ganglion cell layer. Figure 4 In contrast (A, 4C), numerous blood vessels (arrows) and ganglion cell nuclei (triangular heads) are clearly visible. Fluorescence was observed to be located within the ganglion cell layer, while a very small amount of CMP was localized to the internal limiting membrane on the vitreous surface (A, 4C). Figure 4 (B, 4D). In summary, these results indicate that the CMP of the present invention is localized to areas of collagen damage at or near neurons and ganglia, and that this localization leads to ECM repair in these areas, thereby promoting neurite growth and neuronal regeneration, as well as the growth and regeneration of neuronal processes (dendrites and axons), which may ultimately lead to the regeneration and protection of neurons, neuronal processes, and nerves in the peripheral and central nervous systems.

[0516] As another means of examining the effects of CMP treatment on neuronal structure and activity, the ability of CMP to reverse and repair neuronal fragmentation in the corneal nerve bed was investigated. C57BL / 6 mice were bilaterally dried with topical 1% atropine four times daily for a total of 7 days, and on day 7, they were locally treated with either a medium (PBS) or CMP 03A (SEQ ID NO: 1) (200 mM) once daily for an additional 7 days. To assess the degree of neurodegeneration, cholera toxin B (CTB) staining of axons and axonal fragments in various corneal layers was used to determine fragmentation of βIII-tubulin-labeled nerve fibers. The number of continuous neural segments in each subbasal and epithelial terminal image was determined by confocal microscopy. An Olympus FV1000 confocal microscope was used at 800 px. 2 x 800 px 2 Images were acquired z-stacked across the entire central cornea at a resolution of 40x objective lens. Due to the curvature of the flattened cornea, different regions of the same corneal image contain subbasal nerve and epithelial nerve endings in different z-planes, thus requiring separate stacking to compose the final flattened image. To aid segmentation, a Python script was used to segment the image into nine smaller z-stacked patches that could be processed individually. The boundaries of each patch sub-stack were manually selected, and then flattened using the standard deviation z-stack method in ImageJ. This produced individual stacked images of the subbasal plexus and epithelial terminal plexus, respectively. Each image was manually thresholded in ImageJ, and the binary image was saved. The Python script re-stitched the binarized, flattened smaller patches to give the final stacked image of both the subbasal plexus and epithelial terminal plexus. The complete binary image was then processed using a Python script that performed a flooding-fill algorithm on each contiguous region or fragment identified in the binary image. Each unique contiguous fragment was randomly assigned red-green-blue (RGB) values, and images were generated for visual evaluation of neural fragmentation. The number of unique RGB values ​​generated in this way is stored in a record of the number of fragments in each image. A representative pseudo-color fragmented image is shown in... Figure 5 Figure A shows that each consecutive neural segment is identified by a unique color. In untreated mice, the subbasal nerve fibers in the central spiral region are formed by long, continuous segments. Figure 5 A, ascending left). Similarly, although the epithelial terminals are numerous, they remain intact; each nerve ending is mostly monochromatic ( Figure 5 A, ascending middle and right (illustration)). Within the mediator group, numerous short, continuous neural segments are present in the subbasal plexus ( Figure 5 A, middle row, left side), indicating discontinuity. Compared to untreated animals, the number of epithelial nerve endings is reduced and highly fragmented ( Figure 5 A, middle row, middle and right (illustration)), indicating widespread variation. Interestingly, CMP treatment reverses this trend; under the basal layer ( Figure 5 A, bottom row, left side) and epithelial level ( Figure 5 On A, bottom row, middle and right side (illustration) , the nerves appear to have greater continuity and far less fragmentation, and exhibit a similar level of fragmentation as untreated mice. Figure 5 A, top row). It was found that CMP treatment significantly improved nerve fiber growth compared to mediators in the central subbasal layer and epithelial layer. Figure 5 B).

[0517] To further examine the effects of CMP on nerve regeneration, an optic nerve compression experiment was performed. This approach is a useful model of traumatic optic neuropathy and has been used to evaluate potential nerve repair and regeneration strategies (Cameron, EG et al., Bio-protocol 10(6) (March 20, 2020), doi: 10.21769 / BioProtoc.3559). Axonal degeneration in this injury model also involves the degradation of matrix collagen associated with robust tissue remodeling (Bernardo-Colon, A et al., Cell Death Dis. 9:1097 (2018); Sharma, TP et al., Mol.Neurodegener. 9:14 (2014)). Mice were subjected to posterior tubular nerve compression, followed by intravitreal injection of either a mediator (DMSO) or fluorescently labeled CMP 13A (SEQ ID NO:6) three days later. Two weeks after nerve compression, longitudinal sections of the optic nerve were taken at the site of injury, and the axonal length of the sections was examined (by cholera toxin B labeling) and CMP localization (by fluorescence microscopy). The sections were also labeled with glial fibrillary acidic protein (GFAP) to locate astrocytes within the sections. The results of these studies were presented in… Figure 6 As shown in the image. Figure 6 As shown in Figure A, axons containing CTB (false-colored white) from mice treated with the medium extended to the compression site (dashed line), but generally did not extend beyond their distal ends toward the brain. In contrast, axons containing CTB from nerve slices from eyes treated with CMP extended beyond the compression site. Figure 6 B, dashed line), and it is also evident even at more distal locations along the nerve ( Figure 6 B, arrow). The repaired axon largely conforms to the localized plaque of CMP ( Figure 6 C, arrow). Scale bar = 200µm ( Figure 6 A) or 100µm ( Figure 6 B, C).

[0518] Next, we quantified the number of CTB+ green axon segments at discrete distances from the compression site (towards the brain). The results were... Figure 7 As shown in the figure. At each location examined, the nerves from the eye receiving CMP 13A (SEQ ID NO:6) exhibited more axonal segments than the nerves from the mediator eye, with some samples having CTB+ segments extending as far as 1 mm distal to the lesion. Figure 7 Even so, in both cohorts, many samples still had very few (if any) axons at the distal end of the compression site, resulting in significant variability. Figure 7 B, illustration). To better estimate the magnitude of the effect caused by CMP13A compared to the median, we consolidated measurements above the median at each location into three intervals: 50–100 µm, 200–300 µm, and 400 µm, further away from the lesion site. For these locations, nerves from the CMP13A-treated eye showed 29%, 39%, and 96% greater abundance of CTB-containing axonal segments, respectively. Figure 7 C, p≤0.02). The intact axonal segments in these nerves were also longer than those in nerves from the mediator-treated eye. The mean length of intact axonal segments in nerves from the CMP 13A-treated eye was 51% greater than the mean length in the mediator cohort: 122.6 ± 5.5 vs. 81.1 ± 8.3 µm (p < 0.001; Figure 7 D). Similarly, the average length of the 25 longest segments in the CMP 13A nerve was nearly 80% larger than that of the mediator nerve: 228.1 ± 12.9 vs. 127.8 ± 10.5 µm (p < 0.001). Figure 7 E).

[0519] discuss

[0520] The physiological importance of the ECM is reflected in its evolutionary age. A key step in the evolutionary transition from single-celled to multicellular organisms around 600 million years ago was the emergence of genes encoding components of the ECM material, which are essential for providing a platform that allows cells to function as a unit and form tissues (1, 2). Their abundance also reflects the diverse physiological roles of the ECM, which provides structural integrity, mediates extracellular signaling, and promotes specialization in all tissues (3). Indeed, as cells differentiate and associate with other cells to form specialized structures, the composition of the ECM scaffold differentiates species to provide tissue-specific support (4). The importance of the ECM in multicellular tissues is evident early in development, as genes encoding highly conserved ECM proteins are expressed in stem cells as early as the 16-cell stage of the developing embryo in many higher eukaryotes (5, 6). During the development of the CNS, the production of the ECM is regulated spatially and temporally to drive neurogenesis, neural cell migration, and axonal growth and guidance (7). For example, in the visual system, ECM-cell communication is required to drive the connection between retinal-derived axons in the optic nerve and terminal areas in the brain (8).

[0521] The ECM is far more than just a matrix for tissue growth and support; it also serves as a biochemical reservoir of signaling molecules, enabling cells and tissues interacting with the ECM to adapt to environmental cues and stressors (1, 9). The synthesis and release of ECM components and cell-ECM communication are integral parts of many biological processes, including stem cell maintenance and differentiation (10), innervation (11), angiogenesis (12, 13), and wound healing (14). Cells respond not only to the chemical composition of the ECM itself but also to its mechanical properties (15). The ECM responds to resident cells in the tissue through biochemical and biomechanical signals, a process known as “dynamic reciprocity” or “bidirectional crosstalk” (16, 17). Bidirectional relaying of ECM-cell signaling occurs during tissue homeostasis and in pathological conditions (1). Thus, during homeostasis, cells must sense and regulate ECM mechanics to promote the structural integrity and healthy function of the ECM itself (15). ECM homeostasis is maintained by cells sensing mechanical loads acting on the tissue and transducing these mechanical signals through the ECM. This mechanosensory process is controlled by ECM proteins, such as collagen and elastin, which are built to withstand and respond to mechanical stretching and strain (15). Thus, the ECM represents a highly conserved, evolutionarily crucial driver of tissue specificity, connectivity, and adaptation.

[0522] Central nervous system and collagen

[0523] The complex interactions between the ECM and resident cells are reflected and illustrated in the CNS, where the ECM accounts for approximately 20% of total brain mass (18). Neurons and glial cells in the CNS dynamically integrate with the ECM to maintain tissue homeostasis. Although neurons express and secrete various ECM components (19), glial cells are indispensable for ECM deposition and help maintain ECM function and integrity as the CNS ages (20). For example, the structure of the ECM in the CNS changes with age to regulate synaptic plasticity (21). The ECM in the CNS mainly consists of fibrin (including collagen and elastin) and glycoproteins (including proteoglycans, glycoproteins, and laminin), which together form a three-dimensional medium through which the complex network of cells communicates. Smaller homopolymers and heteropolymers combine to form supramolecular assemblies with binding domains for growth factors, cytokines, and cell adhesion molecules (22). In this way, the ECM acts as a medium that can not only deliver ligands but also bind and release them.

[0524] Proteoglycans are highly abundant in neural tissue and form the basis of the higher-order ECM structure surrounding cells. Proteoglycans in neural tissue are rich in covalently bound glycosaminoglycans (GAGs) – long chains of charged polysaccharides (sugars). Major GAGs include heparin sulfate, chondroitin sulfate, hyaluronic acid, and keratin sulfate (7). Even so, collagen remains the most prevalent and indispensable component of the ECM. Nearly 30 types of collagen exist, and their most widely accepted role is providing structural and mechanical integrity to tissues. Collagen is the most abundant protein throughout the human body, particularly types I and III (23). Collagen imparts biological stiffness and strength to tissues (including the ECM), thus influencing the extent to which stress deforms tissues and the maximum stress that can be applied before breakdown (15, 23). Several members of the collagen superfamily, particularly collagen I, IX, and XVIII, participate in CNS development, thus playing important roles in neuronal maturation, neural circuit formation, axonal guidance, and synapsis (24–26).

[0525] The ECM of the CNS is not a uniform, homogeneous “ocean” filling the spaces between neurons, blood vessels, and glial components. Instead, it comprises structurally distinct and specialized landscapes whose structures reflect the function of the surrounding components. These begin with the non-fibrous but protein-rich basal layer. The basal layer acts as a macromolecular sieve-like barrier between tissues, protecting cells from unwanted biochemical and biophysical stressors while also providing a medium for intercellular communication (27, 28). The basal layer is produced by microcapillary endothelial cells, astrocytes, and pericytes (29). Biochemically, the basal layer contains four major ECM proteins: collagen IV, laminin, nestin, and heparan sulfate proteoglycan (HSPG) (29). Minor components include fibulin, osteonectin, and netrin-4 (30). It assembles near the cell surface – consisting primarily of interconnected polymers of collagen IV and VII, with regions that bind to laminin and other glycoproteins (31, 32). Collagen IV, produced by endothelial cells, astrocytes, and perivascular cells, forms the major structural scaffold, which other ECM-related proteins can bind to and interact with (28, 31). The basal layer is also present in the peripheral nervous system, but it differs from the CNS basal layer due to the absence of astrocytes.

[0526] Structural proteins, including fibrous collagen, were thought to be present only along blood vessels and meninges in healthy tissues. Furthermore, it was believed that collagen secretion by astrocytes, neurons, and other glial cells in the CNS was suppressed after development (26). However, the presence of collagen in the brain parenchyma of healthy individuals, including neurons, contradicts these hypotheses (33, 34). Similarly, in the aging human brain, both neurons and astrocytes express genes encoding a range of collagen types (35, 36). Although collagen is not a major component, it appears to play a significant role in the structure and physiology of the peripheral neural network (PNN). Densely packed PNNs, once mistakenly identified as artifacts in Cajal's silverstaining, are condensed ECMs surrounding neuronal cell bodies and proximal / intermediate dendrites, often extending to include the initial segment of the axon. In the brain, they are present in multiple regions, including the hippocampus, cerebral cortex, cerebellum, and basal ganglia (35). Synapses in these regions are surrounded by and embedded in the PNN (37). Loss of collagen XIX leads to a reduction in PNN formation in the mammalian telencephalon (38), and collagen in the hippocampal PNN is associated with the formation of long-term memory (39). Upregulation of collagen IV in the PNN is evident after focal cerebral ischemia (40).

[0527] Dynamic Collagen: More Than Just a Scaffold

[0528] Although collagen fulfills its biological role as a scaffold protein in the ECM, it remains highly dynamic throughout its life cycle, interacting with molecularly bound mates by forming complexes known as the collagen interactant group (41). During collagen biosynthesis, interactions with enzymes such as hydroxylases and lysyl oxidases, as well as chaperone proteins, ensure that collagen adopts its triple-helix structure upon integration into the matrix (42, 43). Once fully formed and integrated, collagen interacts with many other ECM components such as fibronectin, proteoglycans, GAGs, and heparin (42). Collagen also binds to a variety of cell surface receptors, including integrin (44), osteoblast receptors (e.g., OSCAR) (45), mannose receptors (46), and discoid domain receptors (DDR) (42, 47). During turnover or degradation, collagen also interacts with matrix metalloproteinases (MMPs), which contribute to its degradation or digestion (48). Therefore, MMP activity must be a tightly controlled process. An imbalance in MMP activity (i.e., an increase or decrease exceeding the natural collagen turnover requirement) during disease or with age can lead to excessive matrix collagen deposition or degradation in the other direction (49, 50).

[0529] Collagen is a multidomain protein with at least one triple-helical domain that may constitute the majority of its structure, as in type I collagen, or only a small proportion, as in type XII collagen (51). Many other proteins associated with the CNS contain collagen-like domains. The conformational state of collagen determines its binding affinity to specific ligands. Some ligands tend to bind to the native triple-helical collagen structure and have no affinity for fragmented or smaller collagen fragments (41, 52). For example, some ligands, including integrins, DDR1 / 2, MMP1, and molecular chaperone heat shock proteins such as Hsp47 (53), bind only to the native helical structure of collagen (41). The collagen-binding site of the DDR receptor recognizes a specific surface triple-helical sequence on fibrous collagen (54), which is thought to be primarily masked in the native structure of collagen to prevent overactivation of downstream signaling cascades (47, 55). DDR1 is activated by both fibrous and non-fibrous collagen (types I to VI), while DDR2 is activated only by fibrous collagen (types I and III) (56). Integrin receptors are located on the outer surface of collagen (57). However, unwinding of collagen chains exposes RGD sequences within collagen that bind to specific types of integrins (e.g., αvβ3) (58–60). Molecular chaperone Hsp47 binds to only a few GXR motifs in collagen, with most HSP47 binding sites located near the N-terminal portion of the triple-helix region (61). Interestingly, MMPs recognize native collagen but possess inherent helicase activity, unwinding the triple-helix structure to expose MMP binding sites for collagen cleavage (62). Other proteins (such as lysyl oxidase) preferentially bind to higher-order structures such as fibrils (63, 64). In cases of collagen denaturation or damage, proteins involved in collagen turnover recognize epitopes exposed in denatured or degraded collagen (41, 48). Similar to amyloid precursor protein (APP), APP binding to collagen I occurs in both native and damaged states and appears to compete with heparin – suggesting overlap between the binding sites of APP and heparin (65).

[0530] Collagen's tightly coiled triple helix structure makes it less antigenic (its ability to bind to antibodies) and largely non-immunogenic (its ability to induce an immune response through antibodies (66)). However, the bioreactivity of collagen with cell surface antigens and receptors is increasingly considered crucial to disease states (41). Changes in collagen structure, such as damage or increased turnover during disease, can alter its antigenicity and immunogenicity (67, 68). Increased collagen turnover and complete collagen damage are evident during the inflammatory phase of disease and may represent important therapeutic targets in neurodegenerative CNS conditions (69).

[0531] Collagenous ECM landscape as a driver of CNS diseases

[0532] The ECM plays a crucial role as a bioactive scaffold for maintaining biophysical stability and structure, and as a mediator of the diffusion and availability of signaling molecules, such as those mediating interactions between axons and astrocytes (3). Cells in the CNS respond to environmental cues by altering the production and excretion of ECM components, including oxygen or nutrient concentrations and biochemical and mechanical transduction signals. In this sense, the regulation of ECM turnover is essential for neuronal function and survival in the CNS.

[0533] The extracellular landscape of the CNS evolves through changes in the activity of ECM-related genes and enzymes during disease and aging (70). Cellular senescence is becoming a key contributing factor to neurodegenerative diseases of the CNS. Cells entering a senescent state possess a secretory proteome, which includes cytokines and chemokines, as well as ECM, which signals to surrounding tissues (76). In fibrotic diseases outside the CNS, ECM, including collagen, can regulate cellular senescence (77). Changes in ECM occur in a variety of CNS diseases, including Alzheimer's disease (71, 72), Parkinson's disease (73, 74), and optic neuropathy, which is of particular interest to the authors (75, 76). Associated with Alzheimer's disease is the presence of amyloid-β (Aβ) peptide containing type XXV collagen (also known as collagen-like amyloid protein component), which affects amyloid fibril elongation (77), and in some populations, this collagen is genetically associated with Alzheimer's disease (78). Finally, type I collagen contains a binding domain to amyloid precursor proteins, which facilitates monocyte recruitment in disease states (79). In contrast, collagen VI expression in the brains of hAPP mice and individuals with Alzheimer's disease may be neuroprotective; increased collagen VI expression in neurons has a protective effect against Aβ toxicity (80).

[0534] Astrocytes in the central nervous system (CNS) are key players in maintaining the extracellular matrix (ECM). During disease and with aging, the physiological state of astrocytes tends to be more reactive, and changes occur in ECM deposition (81). Collagen is intricately involved in this process, including overproduction, degradation, and compositional alterations detectable in pathophysiological samples (70). Changes in collagen structure and / or deposition affect cell signaling and tissue biomechanics, which in turn can alter cellular responses in tissues, thereby driving disease and inflammatory states (69). Collagen contains multiple binding sites that act as ligands for both cell surface receptors and signaling pathways involved in inflammation. Disruption of these binding sites induced by MMPs leads to chronic inflammation (82, 83). Therefore, even subtle changes in the ECM early in disease progression can be overlooked as drivers of neurodegeneration.

[0535] CNS neurodegeneration is often associated with vascular pathologies such as impaired blood flow and leakage of the blood-brain barrier (81). In systemic vascular diseases such as atherosclerosis, damage to the extracellular matrix (ECM) in the vessel wall leads to disease progression (84). Atherosclerosis is also associated with increased vascular stiffness, likely due to the extensive deposition of collagen (84). Recently, the potential role of complement factors C3 and C4, as well as the ECM, in the vascular etiology of neurodegenerative diseases has emerged. The unexpected observation of C3 and C4 deposition in the vessel wall and co-localization with collagen reveals the potential interaction between complement proteins and collagen in disease and with age (85). In CNS diseases such as Alzheimer's disease, early microglia activation is considered an early driver of neurodegeneration (86, 87). Activated microglia promote neuronal phagocytosis and also lead to disruption of the blood-brain barrier (87). One mechanism of microglia activation depends on the binding of C3 to C3 receptors present on the cell surface of microglia (88). Therefore, damaged collagen present in blood vessels due to aging or disease can act as a novel C3 reservoir, which can trigger early microglia activation and drive neurodegeneration.

[0536] The CNS lacks intrinsic regenerative capacity, partly due to microenvironmental factors that increase local inflammation and reactive glial proliferation (89). Pathological changes in ECM collagen may lead to limited regenerative capacity in the CNS through disrupted signal transduction. Collagen is a major component of astrocyte-mediated ECM deposition, which can promote ECM deposition and influence regeneration in the CNS (90). As matrix collagen degrades, glial fiber complexes formed by the overproduction of type IV collagen secreted by reactive astrocytes create a barrier to axonal repair and regeneration in the CNS by concentrating inhibitory molecules (such as proteoglycans and brain signaling proteins) and inducing the migration of inflammatory microglia and other immune cells (91-94). Thus, this hypertrophic glial complex (commonly referred to as glial scars) provides not only a biomechanical barrier to regeneration but also a biochemical barrier (95). In optic degeneration, changes in the collagen arrangement and stiffness in the sclera (which forms the shape of the eye) and the head of the optic nerve, through which axons pass on their way to the brain, progress through increased inflammation (96).

[0537] Compared to the central nervous system (CNS), the peripheral nervous system possesses a much greater capacity for neuronal repair. Molecular signaling pathways, such as integrins, play a crucial role in the spontaneous regeneration of peripheral axons (97). Collagen is also highly upregulated following peripheral nerve injury and is synthesized by Schwann cells and fibroblasts (98, 99). High levels of collagen at the site of peripheral nerve injury can promote important axonal integrin signaling required for regeneration and may indicate that the role of collagen is more significant than previously thought. Regeneration in non-mammalian species such as zebrafish is strongly ECM-dependent; favorable conditions for regeneration are rich in various collagens, including collagen XII, suggesting the pro-regenerative capacity of certain collagen subtypes (100, 101). Current regeneration strategies focus on modulating growth factor signaling, regeneration-related genes, glial-mediated axonal regeneration, cell replacement, and peripheral nerve tissue transplantation (81), all of which have achieved limited success. None of these utilize the complex signaling capabilities of the ECM (and especially collagen).

[0538] Throughout the body, including in the CNS, the biomechanical properties of tissues can influence cell signaling and cell recruitment to sites of injury or disease. Following CNS injury, the biomechanical stiffness of tissues in the brain and spinal cord decreases, which is associated with increased levels of ECM components, including collagen IV and laminin (95). If collagen is damaged to the extent that it alters the stiffness of a local tissue region, a wave of mechanically induced signaling involving other cells may follow. For example, traumatic injury to the brain involves transient rapid angiogenesis, increased vascular permeability, and accumulation of pro-angiogenic factors, possibly released by microglia (102). These same cells may contribute to tissue remodeling after injury through the secretion of proteases. The vascular basement membrane in the brain contains, among other collagens, heparan sulfate proteoglycan collagen XVIII, which contains a 20-kDa anti-angiogenic endostatin fragment (reviewed in (103)). Following injury, the substantial accumulation of collagen XVIII / endostatin is accompanied by an increase in the number of microglia expressing them (104). This collagen XVIII-dependent process may help counteract early angiogenic damage response to limit secondary damage.

[0539] Repairing collagen in the ECM as a CNS therapeutic agent

[0540] Regarding CNS regeneration, as mentioned above, tissue biomechanics can influence the ability of neurons to grow and guide axons. Therefore, maintaining the inherent biomechanical properties of the ECM itself may be a potential therapeutic avenue. Just as damage to the ECM hinders regeneration, therapies that reconstruct the matrix have great therapeutic potential to promote neural repair and regeneration in conditions including traumatic brain injury (105, 106). This is especially true for therapies that utilize or mimic type I and type IV collagen, both of which are known for their extremely low antigenicity and robust bioavailability (107, 108). Since collagen interactomes and tissue reactivity are largely controlled by collagen structure in the ECM, the role of collagen in tissue ECM homeostasis is becoming increasingly prominent. These observations open up potential therapeutic avenues for matrix repair in CNS diseases. Indeed, one promising area is the potential use of the ECM and ECM-derived peptides to improve neuronal regeneration and functional recovery (109). Similarly, implantation of collagen-rich scaffolds after brain surgery reduces microglia activation and inflammatory-related cytokines (110).

[0541] Collagen is characterized by its triple helix structure—a set of three polypeptide chains containing repetitive sequences of glycine-xy triplets, where x and y typically (but not always) represent proline and hydroxyproline (111). Recent work in our lab and others supports the idea that reconstructing triplets damaged by protease activity can repair CNS tissue and promote neuronal survival in a wide range of neurodegenerative conditions. For example, after optic nerve compression, injection of collagen mimic peptides (CMPs) that insert into and reform the damaged triple helix promotes axonal abduction beyond the compression site and lengthens intact axonal segments (112). Similarly, CMP treatment restored functional axonal transport to central brain targets after optic nerve degeneration induced by elevated intraocular pressure (112, 113). This is a crucial finding because the degradation of axonal transport foreshadows radical axonal degeneration (114, 115). CMPs also exhibit protective and reparative effects on peripheral nerve injury, demonstrating their nutritional capacity for dorsal root ganglia (DRGs) attacked by MMP-induced ECM degradation and for corneal nerve beds damaged by surface dryness (81, 113). Type I collagen-rich gels similarly promote neurite extension from DRG explants (116). The reparative effects of collagen segments are not limited to neurons. Segmented sequences of collagen IV, XV, and XVIII promote angiogenesis and tumor cell growth and influence a variety of other cellular activities (117). Synthetic CMPs target areas of collagen destruction associated with skin wounds by reinforcing natural triple helices through insertion into damaged collagen (118, 119).

[0542] Multiple mechanisms exist through which repair of the damaged triple helix in collagen may influence therapeutic outcomes. Due to its inherent structural properties, collagen, along with other components of the ECM, imparts a significant amount of tissue stiffness (120). Interestingly, a stiffer matrix reduces astrocyte reactivity and the likelihood of glial proliferation, suggesting the formation of inflammatory complexes at CNS injury sites, thereby inhibiting regeneration (121). Following these lines of thought, unlike other scarred tissues, the brain and spinal cord exhibit reduced elastic stiffness after acute injury, consistent with increased astrocyte reactivity and even increased inflammation distal to the injury site (95). In this way, the repair of the stiffer matrix by collagen may help reduce inflammatory responses at the macro-environmental level within the neuronal environment. At the micro-environmental level, MMP activity degrading collagen can disrupt binding sites that will subsequently inhibit local inflammatory signaling complexes in other ways. For example, type I and type III (and possibly type IV) collagen contain a high-affinity binding site for LAIR-1 (leukocyte-associated immunoglobulin-like receptor), which is expressed by most hematopoietic cells to attenuate its activation (122). The intact triple helix in collagen crosslinks with LAIR-1 to inhibit immune cell activation, while the reduced LAIR-1 binding site in disrupted collagen has the opposite effect (82). Interestingly, the sequence of the LAIR-1 (also known as CD305) binding site is conserved between collagen and the complement component C1q, which is an initiator of the classical complement pathway at the root of innate and adaptive immunity (123). C1q mediates the elimination of synapses from dendritic spines in neurons and is activated early in diseases such as Alzheimer's (124). Collagen and C1q mates locally regulate LAIR-1 to avoid immune dysfunction. Similarly, intact type I collagen inhibits the secretion of interleukin-8 from neutrophils through interaction with integrin receptors (125). As collagen chains shorten due to degradation during disease or injury, the threshold for a local inflammatory response is exceeded (83).

[0543] In summary, the CNS ECM is not merely a passive scaffold supporting tissue; it is a diverse, dynamic, and highly bioactive substrate that plays a role in cellular biomechanics and differentiation, as well as cell signaling. While ECM collagen has different roles in tissues outside the CNS, its indispensable role in the CNS during homeostasis and disease has been largely overlooked. Here, we highlight the multiple roles of collagen in the CNS, both at the macroscopic level (in terms of tissue support and biomechanics) and the microscopic level (ligand binding capacity). The structure of collagen is important in its highly functional aspects. Damage to CNS collagen is evident in disease and with aging, and is significant in driving neurodegeneration through its influence on inflammatory pathways. Repairing damaged collagen in the CNS using peptide mimics represents an exciting new therapeutic approach for neurodegenerative diseases, where restoring collagen structure at the microscopic level can help repair, protect, and even regenerate CNS neurons. In summary, these results suggest that the CMP of this invention can be used to enhance the repair of damaged collagen matrix, thereby promoting proliferation, migration, and network formation in neuronal cells. Therefore, the CMP and CMP-containing compositions provided by this invention, as well as their methods of use, should be proven to be useful for the treatment, improvement, prevention, and diagnosis of various diseases and conditions involving nerve cells and the nervous system.

[0544] References

[0545] Example 2: The role of CMP and CMP-TC conjugates in the treatment of cardiovascular diseases

[0546] To examine the potential therapeutic effects of the CMPs of the present invention in the treatment of cardiovascular diseases, studies were designed to test certain CMP conjugates in an in vivo environment for the treatment and possible reversal of atherosclerotic plaques in laboratory animals.

[0547] These studies used an in vivo model of diabetes, namely ApoE. - / - Mice were known to have not only diabetes but also atherosclerotic plaques in their arteries, particularly in the aortic arch. Wild-type or mutant ApoE- / - mice were obtained from Jackson Laboratories (Bartholomew's Harbor, Maine) and were provided with free access to food and water. Mutant mice were treated with streptozotocin (STZ), a compound that has preferential toxicity to pancreatic β-cells and accelerates the progression of atherosclerosis (Rerup, CC, Pharmacol. Rev. 22:485-518 (1970); Szkudelski, T et al., Physiol. Res. 50:537-546 (2001); Rosini, AA et al., Proc. Natl. Acad. Sci. USA 74:2485-2489 (1977)) at 50 mg / kg / animal or with a medicament (PBS) for 5 days. One week after STZ treatment, wild-type and mutant animals were placed on a high-fat diet, and then, starting 7 days later, were injected three times weekly for 10 weeks via tail vein with CMP 03A (SEQ ID NO:1) or PBS. Following this treatment, mice were injected twice more via tail vein with either the medium or fluorescently labeled CMP 03A (100 µl of 100 µM solution) (48 hours apart), and sacrificed 48 hours after the second injection. Animals were perfused with fixative, and liver sections from each mouse were frozen at 10 µm. The perfused liver sections were mounted with DAPI mounting solution (to visualize the nuclei) and imaged at 20x using 405 nm (for DAPI) and 488 nm (for fluorescently labeled CMP) channels. The results of these studies are presented in [Journal Name]. Figure 8 As shown in the image. DAPI staining ( Figure 8 (A1-A3) showed intact hepatocytes with prominent vascular lumens (arrows). Simultaneous evaluation of DAPI and fluorescent CMP (…). Figure 8 (B1-B3) indicates the presence of diffuse CMP fluorescence in wild-type mice. Figure 8 B2), which is mainly located around the vascular lumen (where the collagen-containing basement membrane undergoes constant turnover), while those from ApoE - / - Sections of mutant mice showed numerous punctate CMP fluorescence not only around blood vessels but also throughout the liver parenchyma. Figure 8 The presence of B3 indicates significant collagen damage within the liver tissue itself. These results were confirmed when individual CMP fluorescence was observed in these same sections. Figure 8 (C2, C3). In fact, from Figure 8High-power views of the C2-C3 slice insets show that the CMP is located in the region of expected liver damage associated with the atherosclerosis model. Figure 8 (D3, arrow), the manner of which was not observed in liver sections from wild-type mice ( Figure 8 D2). Further examination of insert sections from mutant mice at even higher magnification ( Figure 9 This confirms the localization of CMP, indicating that collagen damage occurs not only in the vascular basement membrane (arrow) but also within the liver parenchyma (triangle head).

[0548] When the autopsy was performed on wild-type and ApoE - / - These results were confirmed in the mouse liver when the aortic arch of the mutant mice was examined and the localization of fluorescent CMPs was verified. Figure 10 As shown, the aortic arch from wild-type mice only showed diffuse staining, indicating that CMP binds to collagen in tissues that may be undergoing normal turnover, resulting in a diffuse and weak staining pattern. Figure 10 A). In contrast, from ApoE - / - Aortic arch dissected by mice ( Figure 10 B) demonstrated that CMP exhibited significant and differentiated localization within the vessel wall of typical plaque accumulation areas in atherosclerotic animals. This pattern was confirmed in additional sections. Figure 11 These sections not only showed increased accumulation of CMP in the presumed atherosclerotic region of the aortic arch in mutant mice, but also... Figure 11 B), the manner of which was not observed in wild-type mice ( Figure 11 A), and also showed CMP accumulation in vascular areas with significant vascular occlusion – an indication of collagen damage ( Figure 11 B, arrow).

[0549] Finally, check the wild type and ApoE. - / - The localization of the retinal vascular system and neural processes in mice relative to the CMPs of blood vessels and glial astrocytes in these regions. Results in Figure 12 As shown in the figure. In wild-type mice ( Figure 12 (A, 12B) CMPs are diffusely distributed throughout the retinal tissue (arrows), particularly in areas near retinal vessels, a characteristic feature of normal ECM turnover and intrinsic remodeling as described above. In contrast, sections from STZ-treated ApoE- / - mice show significant accumulation of CMPs in localized areas of retinal vessels. Figure 12C, 12D (arrows), particularly in those areas showing extensive glial astrocyte infiltration, presumably due to inflammation in these areas. Therefore, in addition to affecting liver tissue and the vascular system, diabetes and associated atherosclerosis in this mouse model significantly induced the accumulation of CMPs in the perretinal region, suggesting damage to retinal collagen in the optic nerve region.

[0550] These results indicate that damaged collagen accumulates in plaque-developing areas and other tissues (aorta, retina) in a mouse model of atherosclerosis. These findings are consistent with results reported by others, suggesting that collagen damage is a sign of early atherosclerotic development (Smith, KA et al., J. Cardiovas. Transl. Res.16:463 (2023)). Given that CMPs are known to repair damaged collagen in a variety of tissues in humans and other mammals, as well as well described herein, these results suggest that CMPs hold promise as a potential therapeutic agent for treating and potentially reversing atherosclerosis by repairing collagen damage, which appears to be the basis for the development and progression of atherosclerotic plaques. CMPs also appear to be localized in a highly localized manner in the retina of atherosclerotic mice, suggesting their potential for treatment and prevention of certain eye diseases associated with atherosclerosis and diabetes, such as diabetic retinopathy. Furthermore, given that certain central nervous system disorders (e.g., dementia) share some risk factors with atherosclerosis (Nordestgaard, LT et al., Int. J. Mol. Sci. 23:9777 (2022)), CMP treatment can be used to treat both neurological and cardiovascular disorders simultaneously.

[0551] Example 3: The role of CMP and CMP-TC conjugates in the treatment of vascular diseases of the central nervous system

[0552] Neurodegenerative diseases and conditions of the central nervous system (CNS) are often associated with cerebrovascular function. Impaired cerebral blood flow due to vascular remodeling (often due to diseases such as hypertension and atherosclerosis) accelerates cognitive decline and the progression of neurodegenerative diseases such as Alzheimer's disease and dementia (1). Vascular remodeling is an adaptive process that occurs in response to physiological and pathophysiological changes in the vascular microenvironment (2). The vascular microenvironment contains a variety of extracellular matrix (ECM) proteins and certain ECM-degrading proteases, which work synergistically to regulate cellular physiological and pathophysiological processes (2). Collagen is indispensable to the ECM, providing structure and stability to a variety of tissues, including the vascular system (3). Collagen I and III are most abundant in the vessel walls, while collagen IV is a major component of the basement membrane, an integral part of the blood-brain barrier. Endothelial cells (ECs) line the inner surface of blood vessels and act as a barrier between the blood and the vessel walls. ECs become dysfunctional in early vascular diseases, triggering large-scale vascular remodeling, which sometimes precedes the eventual neurodegeneration in the CNS. Vascular remodeling is also a hallmark of atherosclerosis, the most common cause of cardiovascular disease worldwide (4). Remodeling in atherosclerosis is associated with the destruction of collagen within the extravascular membrane (ECM) (5). In the later stages of atherosclerosis, vascular cells, including ECs, secrete increased levels of matrix metalloproteinases (MMPs), leading to proteolytic cleavage of collagen and destabilization of atherosclerotic plaques, thereby increasing the risk of vascular rupture and thrombosis (6). Finally, diabetic patients often exhibit accelerated progression of atherosclerosis due to early EC damage and dysfunction (7, 8).

[0553] The results of the foregoing embodiments and other works published by the inventors (3, 9, 10) have highlighted the potential use of collagen mimic peptides (CMPs) to repair collagen and combat certain neurodegenerative processes. As mentioned above, CMPs are known to interact with and anneal damaged collagen to restore the helical structure of this important ECM component (11-13). In this embodiment, the inventors conducted a study using an established atherosclerosis model to explore the use of CMPs in maintaining vascular architecture in systemic cardiovascular disease, a model that uses streptozotocin (STZ) to accelerate EC dysfunction in ApoE- / - (apolipoprotein E deficient) mice maintained on a high-fat diet (“HFD”) (14, 15). This mouse strain is most widely used in preclinical studies of atherosclerosis, as described in Example 2. The lack of endogenous ApoE in these mice led to the secretion of cytokines and proteases, which subsequently resulted in inflammation and extracellular matrix degradation (16, 17), while systemic STZ application induced a form of type 1 diabetes by reducing endothelial-dependent vasodilatory responses and destroying pancreatic β cells (18, 19).

[0554] Materials and methods

[0555] Animals. All animal studies were conducted in accordance with NIH guidelines for the care and use of laboratory animals and approved by the Vanderbilt University Institutional Animal Care and Use Committee. For the studies described, 12-week-old ApoEs were obtained from Jackson Laboratories. - / - (#002052) or C57 / B6J wild-type (WT) (#000664) mice. Mice were housed in a facility managed by the Vanderbilt University Animal Care Department, with free access to water and a standard diet, followed by free access to a 45% high-fat diet (Research Diet Inc.; catalog number #D12451) starting at the appropriate study time. Mice were subjected to a 12-hr light / dark cycle.

[0556] An animal model of diabetic atherosclerosis. This was achieved through ApoE... - / - A mouse model of diabetic atherosclerosis was established by combining mice with a streptozotocin (STZ)-induced diabetic phenotype. Baseline fasting blood glucose and body weight were obtained before STZ injection and dietary changes. To induce hyperglycemia, ApoE was administered for five consecutive days. - / - Mice were intraperitoneally injected with STZ (50 mg / kg in 10 mM sodium citrate, pH 4.5; Millipore Sigma catalog number #S0130). During the duration of STZ injections, mice were given drinking water supplemented with 10% sucrose. One week after the last STZ injection, blood glucose levels were measured to confirm hyperglycemia. Mice were fasted for 6 hours prior to glucose measurement and then anesthetized with 2.5% isoflurane. Tails were clipped, and a drop of blood was placed on a test strip to obtain a reading. Hemostasis was achieved using a silver nitrate applicator (Avoca reference number #7482), and the mice were returned to their cages. Any mice not registered as hyperglycemic were removed from the study. The remaining mice were then switched to a 45% high-fat diet (HFD) to accelerate atherosclerotic plaque formation. After one week of HFD administration, mice were randomly assigned to receive intravenous CMP (n=5) or PBS (1x PBS, n=4) weekly for 12 weeks. After the study, mice were anesthetized by intraperitoneal injection of pentobarbital, and then perfused with 1X PBS and 4% paraformaldehyde (PFA) via the heart. One day before sacrifice, mice were injected via tail vein with sterile 1X PBS containing sulfonyl-NHS-biotin (Thermo-Fisher; catalog number #21335).

[0557] CMP was administered intravenously. The CMP used in this example is a 21-aa single-chain peptide composed of seven repeating sequences of a tripeptide of 4-fluoroproline (Flp), hydroxyproline (Hyp), and glycine (Gly), abbreviated as (cis-Flp-Hyp-Gly)7 (SEQ ID NO:6). This CMP was manufactured in limited quantities by Bachem, AG (Germany) using standard solid-phase peptide synthesis chemistry. CMP was dissolved in sterile 1X PBS at a concentration of 1 mg / kg and sterilized using a 0.22 μm filter (Millipore Sigma, Burlington, Massachusetts, USA). Mice were weighed and secured in devices designed for tail vein injection. Prior to weekly injections, the tail was heated to dilate the vein, and 100–150 μL of CMP in sterile 1X PBS (the medium) was injected to achieve a final concentration of 0.1 mg / kg per animal. Further batches of this CMP were produced by linking TideFluor™ 2 (AAT Bioquest, Sunnyvale, California, USA) to a peptide that serves as a fluorescent reporter molecule. The labeled CMP was administered intravenously at a concentration of 100 µM (100 μL) to identify WT and ApoE. - / - + Binding in STZ mice. CMP was injected 48 hours prior to sacrifice and immediately retriggered before cardiac perfusion of 4% PFA. One day prior to euthanasia, sulfonyl-NHS biotin was injected into the tail vein to track vascular leakage. Sulfonyl-NHS biotin (Thermo-Fisher; catalog #21335) was dissolved in sterile 1X PBS (20 mg / mL) and sterilized through a 22 µm filter. Mice were then secured in a tail vein apparatus and 100 µL of sulfonyl-NHS biotin was injected intravenously. Organs of interest (spleen, kidney, lung, liver, heart, retina, and brain) were frozen sections at a thickness of 10 µm and fixed on microscope slides for imaging with DAPI Fluoromount-G counterstain (catalog #0100-20; SouthernBiotech, Birmingham, Alabama, USA).

[0558] Immunohistochemistry and fluorescence imaging. After 24 hours, the perfused brain was removed from 4% PFA and cryoprotected in a sucrose gradient series (20% to 30%). The brain was sectioned into 50 µm sections using a cryostat (SM2000R; Leica Biosystems, Buffalo Grove, Illinois, USA), and sections containing the hippocampus were identified for immunolabeling. The tissue was cryoprotected in a sucrose gradient and permeabilized with three freeze-thaw cycles. Sections were washed in 1X PBS for 10 minutes and further permeabilized overnight in 0.5% Triton X-100 in 1X PBS at 4°C. Sections were blocked at room temperature (5% normal donkey serum in 0.1% Triton X-100 in 1X PBS) with agitation for 2 hours, followed by incubation with primary antibodies. Antibodies against CD31 (rat 1:200; BD Pharmingen, 550274), collagen IV (goat 1:100, EMDMillipore AB769), and GFAP (goat 1:200; Abcam, Cambridge, UK, catalog number #ab53554) or the isohemagglutinin GS-IB4 biotin-XX conjugate (1:200; Invitrogen; Waltham, MA, USA, catalog number #I21414) were incubated with shaking at 4°C for 3–5 days in 3% normal donkey serum in 1X PBS and 0.1% triton. Sections were washed with 1X PBS and placed in a secondary antibody solution (1% donkey serum and 0.1% Triton in 1X PBS) containing the following secondary antibodies: donkey anti-rat Alexa 488 (1:200; Jackson Immuno Research; 712-545-150), donkey anti-goat Alexa 555 (1:200; Jackson Immuno Research; catalog number #705-565-147), and streptavidin Alexa-647 (1:200; Invitrogen; catalog number #S21374). Streptavidin binds to sulfonyl-NHS-biotin for fluorescent visualization of injectable biotin. Sections were incubated at room temperature for 2 hours and then washed in 1X PBS. The slides were fixed onto microscope slides (Diamond White Glass Microscope Slides; frosted white; #1358W) using DAPI Fluoromount-G (catalog number #0100-20, SouthernBiotech, Birmingham, Alabama, USA) and the coverslip (microscope cover glass; Globe; #1414-0) was sealed with nail polish (Ted Pella In; #114-7) for imaging.Fluorescence images of the retina and brain were acquired using a Nikon Ti-E rotating confocal microscope. A 10x montage of the brain was taken to orient the CA1 region of the hippocampus, the dentate gyrus (DG), and the cerebral cortex, followed by closer region-specific images at 20x or 60x magnification. Z-stacks of equal thickness were taken in 0.3µm steps. Z-stack images were combined using the standard deviation Z-stack option in Fiji ImageJ. For all other tissues (lung, kidney, liver, spleen, and heart), 20x and 10x magnifications were used on a Nikon NiEclipse fluorescence microscope.

[0559] Quantitative and statistical analyses were performed. Mean fluorescence intensities of GFAP, CD31, collagen IV, and sulfonyl-NHS biotin immunofluorescence were obtained in the CA1 region of the hippocampus, the dentate gyrus (DG), and the cerebral cortex using measurement tools in Fiji software. CD31-stained images were thresholded and binarized. Vessels were then located and shape descriptor information, including area, perimeter, major and minor axes, and Feret diameter, was captured using ImageJ's "Particle Analysis" 30-infinity tool. The distance from astrocytes to vessels was measured using GFAP-stained images of the CA1 and DG brain regions. The cortex was not included in the analysis due to the lack of uniform astrocyte density. Individual astrocyte bodies were located and their centroids were measured using the "Particle Analysis" 30-infinity tool. Minimum distances from each centroid to the nearest vessel were recorded using a custom Python script. All quantifications were performed on slices passing through multiple markers in each brain (typically 2-3 per animal); 12 confocal images were acquired from each brain at 60x magnification, four each for the cortex, CA1, and DG. Statistics were performed in GraphPad Prism version 10.1.2. After testing for normality, ordinary one-way ANOVA, Brown-Forsythe one-way ANOVA, and nonparametric tests including those by Kruskal Wallis and MannWhitney were used.

[0560] result

[0561] 1. Intravenously injected CMP crosses the blood-brain barrier. To determine whether CMP crosses the blood-brain barrier, we intravenously injected fluorescently labeled CMP into healthy WT and diseased ApoE-STZ mice. CMP binding was detected in the cerebral cortex, CA1 region of the hippocampus, and dentate gyrus (DG) region of the brains of WT and ApoE-STZ mice, as evidenced by the regional localization of punctate fluorescent CMPs in areas of damaged collagen. Figure 13 ; arrow). However, the pattern of CMP binding in the brains of ApoE-STZ mice ( Figure 13 D-13F) and the uniform binding observed in WT animals ( Figure 13 The A-13C is different and more ordered. These results indicate that intravenously injected CMP crosses the blood-brain barrier and localizes to regions of disordered collagen that are elevated in animals with diabetic atherosclerosis.

[0562] 2. CMP increases endothelial CD31 and collagen IV expression and helps protect the cerebral vascular architecture. To assess vascular structure and EC density, brain slices were immunolabeled with an antibody that binds to PECAM-1 (CD31) (a potent EC and vascular marker). Figure 14 As shown, in all brain regions, compared with the control group treated with PBS ( Figure 14 Compared to A-14C, CD31 markers were more prominent in animals treated with CMP. Figure 14 D-14F). After quantifying the area and intensity of CD31 staining in various brain regions of mice treated with PBS and CMP, the area of ​​CD31-positive vessels was significantly increased in CMP-treated mice compared to mice treated with PBS, in both CA1 and the cortex. Figure 15 A; p=0.001 and p<0.0001, respectively. Changes in CD31 area were region-specific; for example, there was no significant difference in vascular area in the DG between CMP-treated animals and controls. Interestingly, in all three brain regions of interest, CD31 intensity was increased in CMP-treated mice compared to PBS-treated controls. Figure 15 B): Hippocampus (CA1), cortex, and dentate gyrus (DG) (p=0.02, p<0.0001, and p=0.01, respectively). These results indicate that CMP treatment enhances the organization and angiogenesis (or maintenance of structural integrity) of the EC in the brains of animals with diabetic atherosclerosis.

[0563] To further elucidate the effects of CMP treatment on cerebral vascular integrity, we next evaluated vascular leakage and vascular structure in these animals. For these studies, biotin was intravenously injected into cells treated with PBS and CMP, followed by detection using fluorescently labeled streptavidin. Representative images of biotin labeling in the CA1 cortex and DG are shown in [images]. Figure 16 As shown in the figure. At this time point (3 months post-treatment), no biotin leakage was observed in the vascular system of mice treated with PBS or CMP. However, biotin labeling highlighted differences in vascular structure between these groups. Specifically, in mice treated with PBS ( Figure 16 In A-16C, blood vessels were shorter and more fragmented in all brain regions, while in mice treated with CMP ( Figure 16D-16F exhibited longer, more complex vessels with increased branching. When biotin intensity was quantified as a secondary measure of vascular density, CMP-treated animals showed increased biotin markers in CA1, DG, and cortical areas compared to PBS-treated controls. Figure 17 A). Finally, we evaluated the CD31 intensity as a ratio of biotin intensity. Figure 17 Could the increased CD31 intensity observed in A be due to increased vascular density? Figure 17 B). Interestingly, when biotin intensity was taken into account, CMP treatment only significantly increased vascular CD31 intensity in DG (compared to PBS treatment), suggesting that the increased biotin intensity observed in most brain regions of CMP-treated animals and PBS-treated controls (except DG) is not necessarily due to an increase in the number of blood vessels, but more likely due to an increase in the structural integrity of blood vessels present in CMP-treated animals relative to the mediator-treated controls.

[0564] We next aimed to assess the health of the vascular system in the brains of CMP-treated ApoE-STZ mice and negative controls by determining the expression levels of type IV collagen in the brains of these animals. As mentioned above, type IV collagen is enriched in the basement membrane of intact blood vessels in all regions of the body, including the cerebral vascular system. Therefore, to assess the effect of CMP treatment on vascular collagen IV levels in the brains of ApoE-STZ mice, we labeled brain slices from animals treated with PBS or CMP with antibodies targeting collagen IV. Figure 18 Animals treated with vectors were observed (). Figure 18 Collagen IV strength in A-18C was lower in the cortex, CA1, and DG. However, in the CMP treatment group ( Figure 18 In all brain regions assessed (D-18F), collagen IV showed higher intensity and clearer vascular structures. Figure 19 As shown, compared with the negative control treated with PBS, CMP treatment increased the collagen IV intensity in the cortical brain regions by 175% (p=0.01). Figure 18 Compared with 18D, A increased the intensity of this effect in the CA1 brain region by 170% (p=0.01). Figure 18 B compared to 18E), and increased this intensity in the DG brain region by 52% (p=0.04). Figure 18 C compared to 18F).

[0565] In summary, these results provide compelling evidence to support the conclusion that CMP treatment enhances the organization of ECs and the formation (or maintenance of structural integrity) in the brains of animals with diabetic atherosclerosis.

[0566] 3. CMP treatment promotes astrocyte-vascular interactions in the brain. To evaluate the effects of CMP treatment on glial-vascular interactions in the CA1 and DG brain regions, we quantified the density and number of astrocytes in the brains of ApoE-STZ mice treated with CMP and PBS, as well as their proximity to blood vessels, using a fluorescently labeled anti-GFAP antibody. The cortex was excluded from evaluation due to the lack of GFAP staining (see [link to evaluation]). Figure 13 A, 13D). In animals treated with PBS and CMP, astrocytes ( Figure 20 A, 20C, 20E, and 20G) and their vascular interactions ( Figure 20 Representative images of B, 20D, 20F, and 20H are shown in Figure 20 As shown in the figure. After quantifying the staining intensity, mice treated with CMP showed significantly higher staining intensity in the CA1 region compared to PBS. Figure 21 A(i); p=0.07) and DG region ( Figure 21 There was no significant difference in GFAP density in B(i); p=0.06). Next, we identified and counted individual astrocyte cell bodies in representative microscopic fields. Compared with the PBS control, the number of astrocyte cell bodies in the CA1 region of CMP-treated mice was reduced ( Figure 21 A(ii); p=0.02), while the number of astrocyte cell bodies in DG did not differ significantly between the CMP treatment group and the control group ( Figure 21 B(ii); p=0.31). Finally, using astrocyte cell body coordinates, we calculated the distance from a single astrocyte to the nearest blood vessel in brain slices from mice treated with CMP and PBS. Interestingly, although the number of astrocytes in CA1 was reduced in the CMP-treated group, it was still higher in the CA1 region ( Figure 10 A(iii); p<0.0001) and DG region ( Figure 10 In B(iii); p=0.0004), astrocytes were closer to blood vessels than in mice treated with PBS. These results suggest that CMP treatment promotes increased interaction between helper astrocytes and blood vessels in the brains of animals with diabetic atherosclerosis.

[0567] discuss.

[0568] In our previous work, we demonstrated the potential of CMP to repair collagen in order to counteract neurodegeneration during the projection of the optic nerve into the brain (3, 9, 10). Diabetes and atherosclerosis are systemic diseases associated with age-related neurodegeneration of the CNS, including Alzheimer's disease, diabetic retinopathy, and dementia (21–24). Since vascular dysfunction plays a prominent role as an early driver of neurodegeneration (25), we explored whether CMP has the potential to repair collagen in an accelerated diabetic atherosclerotic disease mouse model (fed HFD on ApoE). - / - Potential for protecting the cerebrovascular system in STZ mice (26, 27, 28).

[0569] Mice fed a normal diet but lacking ApoE exhibited an imbalance of cholesterol in macrophages, leading to high levels of cytokine and protease secretion and triggering inflammation and ECM degradation (17). Feeding ApoE- / - mice a high-fat diet resulted in accelerated atherosclerotic plaque formation and widespread inflammation, accompanied by increased adhesion molecule expression and leukocyte recruitment, as well as microvascular degeneration and reduced microvascular length (16). Changes in vascular structure were associated with activation of the pro-inflammatory cyclophilic protein A (CypA)–nuclear factor-κB–matrix metalloproteinase-9 (MMP9) pathway (29). MMP9 activation leads to collagen degradation in the ECM, which may promote inflammation and vascular degradation (30). STZ is an antibiotic that causes pancreatic β-cell destruction, thereby triggering hyperglycemia (19). When STZ was combined with ApoE deficiency, mice exhibited severe endothelial cell dysfunction (18).

[0570] To investigate the effects of CMP treatment on the pathophysiology of ApoE-STZ (as a model of diabetic cerebrovascular pathology), we treated mice with intravenous PBS or CMP for 12 weeks and assessed vascular structures in the brain. All mice included in the study became hyperglycemic after STZ treatment compared to baseline measurements. CMP crosses the blood-brain barrier (…). Figure 13 It also has a positive effect on the cerebral vascular system. Compared with animals treated with PBS, CMP treatment increased the expression of CD31 in the cortex, CA1, and DG. Figure 14 and 15 Furthermore, compared to the negative control, mice treated with CMP also showed an increased vascular area (assessed by CD31 area and biotin strength). Figure 15-17 After normalization relative to biotin area, CMP treatment resulted in the highest increase of CD31 in DG ( ). Figure 18 , 19 Similarly, CMP treatment was observed to increase collagen IV expression in all brain regions. Figure 20CMP also promotes astrocyte-vascular interactions; compared to PBS treatment, astrocyte cell bodies are more closely aligned with blood vessels after CMP treatment. Figure 21 ).

[0571] Injection of fluorescent CMP confirmed that the mimic peptide crossed the blood-brain barrier and reached the cortex and hippocampus of the brain. Interestingly, CMP binding was observed in both healthy WT and diseased ApoE. - / - Both were evident in animals, indicating the presence of degraded collagen in tissues of both phenotypes. This observation is not surprising, as baseline collagen turnover within the ECM and vascular basal layer occurs in both healthy and diseased tissues during homeostatic processes such as tissue remodeling. ECM turnover is a dynamic but tightly controlled process; at homeostasis, there is a balance between protein degradation and formation (3, 37, 38). However, in disease, this balance is disrupted with increased MMP expression, leading to increased fragmentation of collagen with exposed ligand-binding sites and the production of degraded collagen, which may be involved in multiple pro-inflammatory pathways (3).

[0572] When assessing the effects of CMP on the vascular system in the brain, we first quantified the levels of the EC and the vascular marker CD31 (39). CD31, or platelet / endothelial cell adhesion molecule-1 (PECAM-1), is a cell adhesion molecule highly expressed on the surface of the EC and expressed to a lesser extent by a range of immune cells, including monocytes, neutrophils, and certain T cell subsets. CD31 is an integral component of the microvascular barrier, forming tight junctions at the EC boundaries that mediate vascular permeability and leukocyte migration (39). In the cortical and hippocampal regions of the brain we explored, CMP treatment increased CD31 expression relative to the negative control; however, the upregulation was most pronounced in the vascular system of the DG. Furthermore, in the mediator-treated animals, collagen IV levels were highest in the DG compared to the cortex and CA1, where collagen IV levels were much lower. In vascular dementia, the effects of ischemic injury on brain regions are known to vary. For example, unlike other brain regions, DG exhibits compensatory mechanisms in response to ischemia, such as synaptic plasticity, activation of resident glial cells, angiogenesis, and stem cell proliferation (40). In Alzheimer's patients, DG also demonstrates resistance to plaque accumulation (41). The fact that DG is capable of undergoing neurogenesis (42) and exhibits the highest levels of CD31 in CMP-treated animals suggests that helical collagen in DG plays an indispensable role in EC physiology, which may in turn influence neuronal health and neurogenesis. Given the promising results observed here, collagen mimic peptides may be important therapeutic compounds for promoting neurogenesis and cognitive function in humans and mammals.

[0573] At the end of this study (12 weeks), we did not detect significant vascular leakage using intravenous biotin injections. However, we did detect a reduction in vessel length and disease remodeling, which appeared to be prevented by CMP treatment. It is not surprising that vascular structure was preserved, given that CMP binds to collagen and preserves the natural structure of collagen. CMP also increased CD31 expression on ECs, which is essential for maintaining immune immunity of the vascular endothelium and preserving vascular structure and integrity (44,45). Therefore, the improved vascular structure we observed may also be due to the CD31 elevation induced by CMP treatment, again highlighting the potential therapeutic benefit of CMP treatment in this context, as preventing vascular rupture is crucial for maintaining the blood-brain barrier and immune immunity of the brain to prevent neurodegeneration (46). Finally, our results suggest that CMP treatment promotes vascular-astrocytic interactions at the cellular level. Astrocytes are glial cells that, along with endothelial cells, pericytes, and microglia, maintain a tight blood-brain barrier by forming neurovascular units (47). Our results indicate that CMP treatment will help prevent neurovascular rupture and even promote cerebral vascular nerve regeneration and growth, as well as improve the integrity of the cerebral vascular network itself. This is clearly of therapeutic significance for the treatment and / or prevention of CNS diseases (including Alzheimer's disease and other dementias) and PNS diseases (such as amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Huntington's disease), as well as diseases and conditions characterized by blockage, trauma, or leakage of the cerebral vascular system (such as stroke, aneurysm, cerebral hemorrhage, cerebral microbleeds, etc.).

[0574] Collagen is not an inert member of the ECM landscape, but rather acts as a key component of a wide variety of cell signaling pathways (3). The breakdown of collagen within the ECM can promote unwanted cell signaling cascades (e.g., pro-inflammatory or pro-apoptotic) or lead to neurodegenerative changes in cell state. As a hallmark of aging, cellular senescence is a significant contributing factor to aging and age-related diseases, including AD. ECs line the inner walls of blood vessels and are exposed to constant shear stress from blood cell flow (48). ECs located at arterial geometries (such as tortuosity and branching) experience higher cell turnover and thus often become senescent (49). Atherosclerosis and diabetes are both diseases associated with hypertension, which increase shear stress and thus increase cellular senescence in ECs (50, 51). Furthermore, glycation of type I collagen leads to a premature senescence-like state in endothelial cells (52). Senescent cells adopt an inflammatory senescence-associated secretory phenotype (SASP), releasing excessive amounts of pro-inflammatory factors such as interleukin-1β (IL-1β), interleukin-6, and MMPs (49). Therefore, the accumulation of senescent cells in the brain may lead to ECM remodeling and collagen breakdown, thereby triggering inflammatory cell activation and subsequent neurodegeneration (53).

[0575] In summary, the results presented here provide strong evidence for the therapeutic benefits of CMP. Therefore, it can be expected that CMP treatment in humans and mammals will lead to the treatment, prevention, and potential reversal of various cardiovascular and cerebrovascular diseases.

[0576] References .

[0577] The present invention has been described above by means of functional building blocks that illustrate the implementation of specific functions and their relationships. For ease of description, the boundaries of these functional building blocks have not been defined herein. Alternative boundaries may be defined as long as the specific functions and their relationships are properly performed. For example, the expression of a numerical range (e.g., a range of doses or administered concentrations) should be understood to include the beginning and end values ​​of the range, as well as each value between the beginning and end values. To illustrate this concept, the range “about 25 ng / ml to about 250 ng / ml” should be interpreted to include the values ​​of “about 25 ng / ml”, “about 250 ng / ml”, and each individual concentration value between the two values. The term “about” when used in conjunction with numerical values ​​generally means a value ±10% of the actual value.

[0578] The foregoing description of specific embodiments will fully reveal the general nature of the invention, enabling others to readily modify and / or adapt such specific embodiments for various applications by applying knowledge within the scope of the art without departing from the general concept of the invention. Therefore, in addition to the embodiments specifically described herein, other suitable embodiments of the invention will be apparent to those skilled in the art based on the foregoing description and examples and knowledge generally available in the relevant art. Thus, based on the teachings and guidance presented herein, such modifications and alterations are intended to be within the meaning and scope of equivalents of the disclosed embodiments. It should be understood that the wording or terminology herein is intended to be illustrative rather than limiting, and those skilled in the art will interpret the terminology or terminology in this specification in accordance with the teachings and guidance.

[0579] The breadth and scope of this invention should not be limited to the exemplary embodiments described above, but should be defined solely by the appended claims and their equivalents.

[0580] All references cited in this document, including U.S. patents and published patent applications, international patents and patent applications, journal references, or other publicly available documents, are incorporated herein by reference in their entirety as if each reference were specifically cited for the part or parts of that reference applicable to the part relating to this application.

Claims

1. A composition suitable for use in a medicament for treating or preventing a neurological disease, condition, or injury in a human or animal, the composition comprising (a) at least one collagen mimic peptide (CMP), and (b) one or more pharmaceutically suitable carriers.

2. The composition of claim 1, wherein the at least one CMP is linked to at least one therapeutic compound (TC) to form a CMP-TC conjugate.

3. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1-388, 397-416 and 418-478.

4. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1-14, 66-94, 107-135, 136-140, 192-220, 233-261, 260-264, 280, 281, 293, 294, 306, 307, 318-346, 347, 348, 359-388, 397-416, and 418-478.

5. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to SEQ ID NO:

6.

6. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1, 5 or 473.

7. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 9, 388, 397-416 and 418-478.

8. The composition of claim 1, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 10-27, 81-94, 122-135, 207-220, 248-261, 333-346 and 374-387.

9. A method for treating or preventing a neurological disease, condition, or injury in a person or animal requiring treatment or prevention, the method comprising administering to the person or animal a composition according to any one of claims 1-8 at a dose sufficient to treat or prevent the neurological condition or injury, monitoring the state of the condition or injury in the person or animal over time, and re-administering the composition to the person or animal until the neurological condition or injury is cured, repaired, prevented, or improved.

10. The method of claim 9, wherein the method is used to treat or prevent diseases, conditions or injuries of the central nervous system.

11. The method of claim 10, wherein the central nervous system disease, condition, or injury is selected from the group consisting of multiple sclerosis, amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, traumatic encephalopathy, non-Alzheimer's dementia, encephalitis, chronic traumatic encephalopathy, and meningitis.

12. The method of claim 10, wherein the central nervous system disease, condition, or injury is a neurodegenerative disease or condition.

13. The method of claim 9, wherein the method is used to treat or prevent peripheral nervous system diseases, conditions, or injuries.

14. The method of claim 13, wherein the peripheral nervous system disease, symptom, or injury is a peripheral neuropathy or injury in which one or more peripheral nerves are severed or compressed.

15. The method of claim 9, wherein the composition is applied parenterally to the human or animal.

16. The method of claim 15, wherein the composition is administered to the human or animal via intravenous, subcutaneous, intramuscular, intradermal, transdermal, oral, inhalation, rectal, or intrathecal administration.

17. The method of claim 9, wherein the composition is applied topically to the human or animal.

18. The method of claim 9, wherein the composition is applied to the human or animal as a coating on a solid or semi-solid material, the solid or semi-solid material being implanted into one or more tissues or organs of the human or animal.

19. The method of claim 18, wherein the solid or semi-solid material is a sheet, film, gel, mesh, or patch.

20. The method of claim 18, wherein the solid or semi-solid material is one or more spheres or nanoparticles.

21. A medical device suitable for treating or preventing neurological diseases, conditions, or injuries in humans or animals requiring treatment or prevention, wherein the device comprises at least one of the compositions claimed in any one of claims 1-8.

22. The medical device of claim 21, wherein the device is selected from the group consisting of a stent, a shunt, a suture, an absorbable mesh, an absorbable patch, a drug-releasing sheet, a membrane, and an internal infusion pump.

23. A method for treating, improving, or preventing a neurological disease, condition, or injury in a person or animal requiring treatment or prevention, the method comprising implanting a medical device as claimed in claim 21 into one or more tissues or organs of the person or animal, and monitoring the medical condition of the person or animal until the neurological disease, condition, or injury is cured, repaired, improved, or prevented.

24. A composition suitable for use in a medicament for treating or preventing cardiovascular diseases, conditions, or injuries in humans or animals requiring treatment or prevention, the composition comprising (a) at least one collagen mimic peptide (CMP) and (b) one or more pharmaceutically suitable carriers.

25. The composition of claim 24, wherein the at least one CMP is linked to at least one therapeutic compound (TC) to form a CMP-TC conjugate.

26. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1-388, 397-416 and 418-478.

27. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1-14, 66-94, 107-135, 136-140, 192-220, 233-261, 260-264, 280, 281, 293, 294, 306, 307, 318-346, 347, 348, 359-388, 397-416, and 418-478.

28. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to SEQ ID NO:

6.

29. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 1, 5 or 473.

30. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 9, 388, 397-416 and 418-478.

31. The composition of claim 24, wherein the at least one collagen mimic peptide has an amino acid sequence corresponding to any one of SEQ ID NO: 10-27, 81-94, 122-135, 207-220, 248-261, 333-346 and 374-387.

32. A method for treating or preventing cardiovascular disease, condition, or injury in a person or animal requiring treatment or prevention, the method comprising administering to the person or animal a composition as described in any one of claims 24-31 at a dose sufficient to treat or prevent the cardiovascular disease or injury, monitoring the state of the disease or injury in the person or animal over time, and re-administering the composition to the person or animal until the cardiovascular disease or injury is cured, repaired, prevented, or improved.

33. The method of claim 32, wherein the method is used to treat or prevent atherosclerosis.

34. The method of claim 33, wherein the atherosclerosis is diabetic atherosclerosis.

35. The method of claim 32, wherein the cardiovascular disease, condition, or injury is a disease, condition, or injury of one or more blood vessels in the brain.

36. The method of claim 35, wherein the disease, condition or injury of one or more blood vessels of the brain is selected from the group consisting of stroke, aneurysm, cerebral hemorrhage, cerebral microbleed, and chronic traumatic encephalopathy.

37. The method of claim 32, wherein the composition is applied parenterally to the human or animal.

38. The method of claim 37, wherein the composition is administered to the human or animal via intravenous, subcutaneous, oral, inhalation, rectal, or intrathecal administration.

39. The method of claim 32, wherein the composition is applied topically to the human or animal.

40. The method of claim 32, wherein the composition is applied to the human or animal as a coating on a solid or semi-solid material, the solid or semi-solid material being implanted into one or more tissues or organs of the human or animal.

41. The method of claim 40, wherein the solid or semi-solid material is a sheet, film, gel, mesh, or patch.

42. The method of claim 40, wherein the solid or semi-solid material is one or more spheres or nanoparticles.

43. A medical device suitable for treating or preventing cardiovascular diseases, conditions, or injuries in humans or animals requiring treatment or prevention, wherein the device comprises at least one of the compositions claimed in any one of claims 24-31.

44. The medical device of claim 43, wherein the device is selected from the group consisting of a stent, a shunt, a suture, an absorbable mesh, an absorbable patch, a drug-releasing sheet, a membrane, and an internal infusion pump.

45. A method for treating, improving, or preventing cardiovascular disease, condition, or injury in a person or animal requiring treatment or prevention, the method comprising implanting a medical device as claimed in claim 43 into one or more tissues or organs of the person or animal, and monitoring the medical condition of the person or animal until the cardiovascular disease, condition, or injury is cured, repaired, improved, or prevented.

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