Collagen impregnation device and method for cancer treatment

Abstract

Collagen devices and methods for treating cancer in a patient. This method comprises the step of introducing a collagen device having a DDR1 peptide into a patient, thereby delivering the DDRI peptide to the cancer. Other such methods include the step of introducing a DDR1 peptide into a patient, thereby delivering the DDRI peptide to the cancer. These methods may also include the step of administering an anti-cancer therapeutic agent to the patient.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 349,300, filed on June 6, 2022, the entire contents of which are incorporated herein by reference. Technical Field

[0003] Embodiments of the present invention relate to customizable collagens for various tissues for different purposes, including cancer treatment. More specifically, embodiments of the present invention relate to methods for designing a device for releasing peptides over time to treat cancer. The method may include amplifying or reducing certain features of collagen for targeted uses (e.g., for these cancer applications) (e.g., as described in the applicant's international patent application PCT / US2021 / 049646, published as WO 2022 / 060622 A1, the disclosure of which is incorporated herein by reference). Embodiments of the present invention also relate to engineered collagen constructs, including hierarchical targeted collagen products / constructs, methods for preparing them, methods for using 3D printing of biomolecules including customizable collagen, and collagen constructs formed by these methods. Embodiments also include injectable forms and bandage designs. Further embodiments include one or more active pharmaceutical ingredients (APIs) combined with cell-binding peptides, and transforming or inducing anti-cancer immune cells, such as T cells, to express DDR1 peptides, thereby enabling them to prevent collagen arrangement. Background Art

[0004] Collagen is a protein found naturally in humans and animals. Collagen tissue is often obtained from donors and used in a variety of medical applications from plastic surgery to bone repair to wound healing. Prokaryotic collagen can also be derived from genetically engineered microorganisms.

[0005] Collagen can be treated and / or processed in a variety of different ways, such as enhancement, reorganization, concentration, cross-linking, combination with other biological substances, etc. Therefore, various collagen products can be produced for different medical applications. Some embodiments of the present invention generally relate to rapid prototyping systems, and in particular, to 3D printing systems for manufacturing medical and dental devices based on collagen, such as dental bone grafts, dental membranes, stents, punctal plugs, ocular collagen inlays and onlays, contact lenses, orthopedic bone applications, spinal applications, neurological applications, and skin applications. Various embodiments / processes of the present invention involve the use of inkjet printing, fused deposition modeling (FDM), selective laser sintering (SLS), stereolithography (SLA), digital light processing (DLP), bioprinting, or a combination thereof, to construct medical devices into three-dimensional objects from many material systems and novel resin systems of the present invention. The inkjet printing system distributes materials through an inkjet print head to form 3D objects, which are hardened by cooling, polymerization, and light exposure. FDM extrude thermoplastic materials throughout the nozzle to build 3D objects. SLS uses a laser as a power source to sinter powdered materials to form solid objects. SLA uses a laser beam to outline the shape of each layer and hardens the photosensitive resin in a vat (reservoir or bath). DLP systems build three-dimensional objects by projecting successive planes of voxels into a liquid resin using a digital light processor (DLP) projector, which then cures the liquid resin. Bioprinting is a layer-by-layer process in which a biological matrix is ​​printed with or without cells. These objects can then serve as a matrix or scaffold to grow cellularized tissues. In general, rapid prototyping refers to traditional manufacturing processes used to make parts, where parts are built up layer by layer using layers of hardened material. According to this technology, the part to be made is considered to be a series of discrete cross-sectional areas that, when combined together, make up a three-dimensional structure. Building parts layer by layer is very different from traditional machining techniques, which involve cutting and drilling metal or plastic parts into the desired shape. In rapid prototyping techniques, parts are produced directly from a computer-aided design (CAD) or other digital image. Software is used to slice the digital image into thin cross-sectional layers. The part is then constructed by stacking layers of plastic or other hardened material on top of each other. There are many different techniques for combining structural layer materials. For some technologies, a final curing step may be required to fully solidify the material layer. For some technologies, such as inkjet printing with powder bed or FDM, a sealant may be required to form a dense 3D object. Some technologies may add additional grinding.

[0006] Inkjet printing technology is a rapid prototyping method that can be used to make three-dimensional objects. In a well-known inkjet printing method developed at the Massachusetts Institute of Technology, as described in Sachs et al., U.S. Patent No. 5,204,055 (the entire contents of which are incorporated herein by reference), a printer head is used to discharge a binder material onto a layer of powder particles in a powder bed. The powder layer corresponds to the digital superposition of the object to be produced. The binder causes the powder particles to fuse together in selected areas. This results in the formation of a molten cross-sectional segment of the object on the platform. These steps are repeated for each new layer until the desired goal is achieved. In the last step, a laser beam is scanned over the object, causing the powder layers to sinter and fuse together where necessary. In another inkjet printing process, as described in Sanders, U.S. Patent Nos. 5,506,607 and 5,740,051, a low melting point thermoplastic material is dispensed through an inkjet print head to form a three-dimensional object. A second inkjet printer head dispenses a wax material or other support material to form a support for the three-dimensional object. After the object is made, the wax support is removed and the object is completed as desired. MultiJet printers, such as the high-quality PolyJet and MultiJet 3D printing processes, use UV light to cross-link photopolymers. However, instead of scanning lasers to solidify layers, the printer jets eject tiny droplets of photopolymer (similar to ink in an inkjet printer) in the shape of the first layer. UV lamps attached to the printer head cross-link the polymers and lock the shape of the layer in place. The build platform then drops down one layer thick and more material is deposited directly on top of the previous layer. Triple Jetting Technology (PolyJet) used in the Stratasys Objet 500Connex3 is the most advanced method in PolyJet 3D printing. The technology uses three materials for precise printing, resulting in a three-color mix.

[0007] Scott Crump, founder of Stratasys Ltd., first developed and implemented the Fused Deposition Modeling (FDM) technology in the 1980s. The advantage of this technology is that all parts printed with FDM can use high-performance and engineering-grade thermoplastics. FDM is the only 3D printing technology that uses production-grade thermoplastics to make parts, so the printed things have excellent mechanical, thermal and chemical qualities. 3D printers using FDM technology build objects layer by layer from the bottom up by heating and extruding thermoplastic filaments. In addition to thermoplastics, printers can also extrude support materials. The printer then heats the thermoplastic to the melting point and extrude it through a nozzle onto the build platform. To support the upper layer, the printer can place special materials that can be dissolved after printing is completed. When the thin layer of plastic combines with the plastic layer below, it cools and hardens. After the layer is completed, the base is lowered to start building the next layer. This technology is considered simple to use and environmentally friendly. Different kinds of thermoplastics can be used to print dental objects.

[0008] Selective Laser Sintering (SLS) is a technique that uses lasers as a power source to form solid 3D objects. The technology was developed by Carl Deckard, a student at the University of Texas, and his professor Joe Beaman in the 1980s. The main difference between SLS and SLA is that it uses powdered material in a vat instead of liquid resin like stereolithography. Unlike some other additive manufacturing processes such as stereolithography (SLA) and fused deposition modeling (FDM), SLS does not require the use of any support structures since the object being printed is always surrounded by unsintered powder. Due to the wide variety of materials that can be used with this type of 3D printer, this technology is very popular for 3D printing customized products. SLS requires the use of high-powered lasers, which makes the printers very expensive. Dental objects made with this process require extensive surface treatment.

[0009] The SLA 3D printing method was patented in 1986 by Charles Hull, co-founder of 3D Systems, Inc., which transforms liquid plastics into solid 3D objects. SLA 3D printers use an excess of liquid resin, which hardens and forms a solid object through irradiation. The manufactured parts usually have a smooth surface, but their quality varies depending on the quality of the SLA machine used. After the plastic hardens, the printer's platform is lowered (top-down printers) or raised (bottom-up printers) by one millimeter in the tank, and the laser forms the next layer until the print is complete. After all layers have been printed, the object is rinsed with a solvent and then placed in a post-curing oven to complete the process.

[0010] Digital Light Processing is another 3D printing process that is very similar to stereolithography. DLP technology was created in 1987 by Larry Hornbeck of Texas Instruments and is very popular in the production of projectors. It uses digital micromirrors arranged on a semiconductor chip. 3D inkjet, DLP, and SLA all work with photopolymers. The difference between SLA and DLP processes is the different light sources. The DLP method projects a continuous plane of voxels into a liquid resin, which then cures the liquid resin. The material used for printing is a liquid resin placed in a transparent resin container. When exposed to light, the resin hardens rapidly. Printing speeds are impressive, especially with Carbon3D's CLIP (Continuous Liquid Interface Production) technology. Layers of hardened material can be created with this printer in seconds. When the layer is completed, it moves up and begins processing the next layer. CLIP technology balances light and oxygen, eliminating the mechanical steps and layers of the standard DLP process steps and allowing for the high-speed production of commercial-quality objects.

[0011] Bioprinting involves a liquid mixture of cells, matrix and nutrients, called bio-ink, being placed in a printer cartridge and deposited using a patient's medical scan. When the bioprinted pre-tissue is transferred to an incubator, this cell-based pre-tissue matures into tissue. Additionally, the matrix can be printed without cells and then populated with cells in vivo or in vitro.

[0012] 3D bioprinting for making biological constructs typically involves distributing cells onto a biocompatible scaffold using a continuous layer-by-layer approach to produce tissue-like three-dimensional structures. Given that every tissue in the body is naturally composed of different types of cells, the many techniques for printing these cells vary in their ability to ensure the stability and survival of the cells during the manufacturing process. Some methods for 3D bioprinting of cells - some of the printing techniques mentioned above - as well as extrusion printing into a support gel. Summary of the invention

[0013] Various embodiments of methods for treating cancer are disclosed in the patent. A method of treating cancer includes the step of introducing a collagen device having a DDR1 peptide into a patient's body, thereby delivering the DDR1 peptide to the cancer.

[0014] Another method of treating cancer comprises the step of introducing a DDR1 peptide into a patient, thereby delivering the DDR1 polypeptide to the cancer.

[0015] Another method of treating cancer comprises the steps of introducing a DDR1 peptide into a patient, thereby delivering the DDR1 peptide to the cancer; and administering an anti-cancer therapeutic agent to the patient.

[0016] Another method of treating cancer includes the steps of introducing a collagen device having a DDR1 peptide into a patient, thereby delivering the DDR1 peptide to the cancer; and administering an anti-cancer therapeutic agent to the patient.

[0017] Another method of cancer treatment involves administering a therapeutic agent derivatized with a targeting peptide that binds to a receptor present on cancer cells.

[0018] Another cancer treatment method includes administering a therapeutic agent derived with a synthetic receptor or targeting peptide that binds to a collagen device, including DDR receptors, selected integrin receptors (α1β1, α2β1, β10β1, and α11β1), and selected immunoglobulin (IgG)-like receptors, glycoprotein VI (GPVI), leukocyte-associated immunoglobulin-like receptor 1 (LAIR-1), and osteoclast-associated receptor (OSCAR), and introducing the collagen device into the patient's body to deliver the therapeutic agent to the cancer.

[0019] Various embodiments of collagen devices and methods of forming the same are also disclosed.

[0020] Other aspects of the present invention include the following:

[0021] 1. A method for treating cancer in a patient, comprising the steps of:

[0022] A collagen device comprising the DDR1 peptide is introduced into the patient, thereby delivering the DDR1 peptide to the cancer.

[0023] 2. The method according to aspect 1, wherein the collagen device is an injectable device comprising any one of type III collagen, rh type III collagen (modified with DDR1 binding site or native), wherein the type III collagen, rh type III collagen comprises a DDR1 peptide.

[0024] 3. The method according to aspect 1, wherein the collagen device is an inlay device or a film comprising any type III collagen or rh type III collagen (modified with DDR1 binding sites or native), wherein the type III collagen, rh type III collagen comprises a DDR1 peptide.

[0025] 4. A method of treating cancer in a patient, comprising the steps of:

[0026] The DDR1 peptide is introduced into the patient, thereby delivering the DDR1 peptide to the cancer.

[0027] 5. The method according to aspect 1 or 4, wherein the DDR1 peptide or the collagen device comprising the DDR1 peptide is dispersed in a degradable carrier.

[0028] 6. The method according to aspect 5, wherein the carrier is selected from gelatin or other collagen

[0029] 7. The method according to aspect 5, wherein the carrier is selected from synthetic degradable polymers.

[0030] 8. The device according to aspect 7, wherein the carrier is selected from PEG, PLA, PGA, PLGA, polymers thereof, and the like.

[0031] 9. The aspect of claim 1 or 4, wherein the DDR1 peptide dispersed in a carrier or collagen device is injectable, said DDR1 peptide comprising the DDR1 peptide dispersed in a carrier.

[0032] 10. A method of treating cancer in a patient, comprising the steps of:

[0033] introducing a collagen device comprising a DDR1 peptide into a patient's body, thereby delivering the DDR1 peptide to the cancer; and co-administering an anti-cancer therapeutic agent.

[0034] 11. The method of aspect 10, wherein the therapeutic agent is a cancer API.

[0035] 12. The method of aspect 10, wherein the therapeutic agent is a cancer vaccine.

[0036] 13. The method according to aspect 10, wherein the therapeutic agent is an inhibitor / degrader of HPK1.

[0037] 14. A peptide drug conjugate comprising [(therapeutic agent)-(connector peptide)-(DDR1 peptide)], as described in the specific embodiments of the present invention, referred to herein as Construct 1.

[0038] 15. A peptide drug conjugate comprising [(therapeutic agent)-(connector peptide)-(non-DDR1 peptide)], as described in the specific embodiments of the present invention, referred to herein as Construct 2.

[0039] 16. A peptide drug or collagen III receptor drug conjugate, comprising [(therapeutic agent)-(connecting peptide)-(collagen III receptor or targeting peptide)], as described in the specific embodiments of the present invention, herein referred to as construct 3.

[0040] 17. A type I collagen receptor drug conjugate comprising [(therapeutic agent)-(connecting peptide)-(type I collagen receptor)], as described in the specific embodiments of the present invention, herein referred to as construct 4.

[0041] 18. A synthetic cell receptor drug conjugate comprising [(therapeutic agent)-(connecting peptide)-(synthetic cell receptor)], as described in the specific embodiments of the present invention, herein referred to as Construct 5.

[0042] 19. A peptide drug conjugate comprising [(therapeutic agent)-(connecting peptide)-GPRGQOGVMGFO)], as described in the specific embodiments of the present invention, referred to herein as Construct 6.

[0043] 20. A method of treating cancer in a patient, comprising the steps of:

[0044] The conjugate construct 1 is administered into a patient, thereby delivering the conjugate to the cancer.

[0045] 21. A method of treating cancer in a patient, comprising the steps of:

[0046] Conjugate construct 2 is administered to the patient, thereby delivering the conjugate to the cancer.

[0047] 22. A method of treating cancer in a patient, comprising the steps of:

[0048] conjugating the conjugate construct 3 to the collagen device; and

[0049] The collagen device is introduced into the patient's body, thereby delivering the conjugate to the cancer.

[0050] 23. A method of treating cancer in a patient, comprising the steps of:

[0051] Conjugate construct 4 is administered to the patient, thereby delivering the conjugate to the cancer.

[0052] 24. A method of treating cancer in a patient, comprising the steps of:

[0053] conjugating conjugate construct 4 to the type I collagen device; and

[0054] The collagen device is introduced into the patient's body, thereby delivering the conjugate to the cancer.

[0055] 25. A method of treating cancer in a patient, comprising the steps of:

[0056] conjugating the conjugate construct 5 to the collagen device; and

[0057] The collagen device is introduced into the patient's body, thereby delivering the conjugate to the cancer.

[0058] 26. A method of treating cancer in a patient, comprising the steps of:

[0059] The conjugate construct 6 is administered to the patient, thereby delivering the conjugate to the cancer.

[0060] 27. A method of treating cancer in a patient, comprising the steps of:

[0061] attaching the conjugate construct 6 to the collagen device; and

[0062] The collagen device is introduced into the patient's body, thereby delivering the conjugate to the cancer.

[0063] 28. The method of aspect 22, wherein the collagen receptor is selected from the group consisting of DDR1, DDR2, GPVI, LAIR-1 and OSCAR receptor.

[0064] 29. The method of aspect 22, wherein the collagen binding peptide is a sequence from vWF.

[0065] 30. The method according to aspect 29, wherein the collagen binding peptide is selected from

[0066] a. Any peptide containing the sequence GxRGQOGVMGFO.

[0067] b. Any peptide containing the sequence RGQOGVMGF.

[0068] c. Any peptide containing the sequence GPRGQOGVMGFO.

[0069] 31. The device according to aspect 2, wherein construct 1 is dispersed.

[0070] 32. The device according to aspect 31, wherein construct 1 is dispersed in a degradable carrier.

[0071] 33. The device according to aspect 32, wherein the carrier is selected from gelatin or other collagen

[0072] 34. The device according to aspect 32, wherein the carrier is selected from synthetic degradable polymers.

[0073] 35. The device of aspect 34, wherein the carrier is selected from PEG, PLA, PGA, PLGA, polymers thereof, and the like.

[0074] 36. The device according to aspect 3, wherein construct 1 is dispersed.

[0075] 37. The device according to aspect 36, wherein construct 1 is dispersed in a degradable carrier.

[0076] 38. The device according to aspect 37, wherein the carrier is selected from gelatin or other collagen

[0077] 39. The device according to aspect 37, wherein the carrier is selected from synthetic degradable polymers.

[0078] 40. The device of aspect 39, wherein the carrier is selected from the group consisting of PEG, PLA, PGA, PLGA, polymers thereof, and the like.

[0079] 41. The device according to aspect 22, wherein construct 1 is dispersed.

[0080] 42. The device according to aspect 41, wherein construct 1 is dispersed in a degradable carrier.

[0081] 43. The device according to aspect 42, wherein the carrier is selected from gelatin or other collagen

[0082] 44. The device according to aspect 42, wherein the carrier is selected from synthetic degradable polymers.

[0083] 45. The device of aspect 44, wherein the carrier is selected from PEG, PLA, PGA, PLGA, polymers thereof, and the like.

[0084] 46. ​​The device according to aspect 24, wherein construct 4 is dispersed in a degradable carrier.

[0085] 47. The device according to aspect 46, wherein the carrier is selected from gelatin or other collagen.

[0086] 48. The device according to aspect 46, wherein the carrier is selected from a synthetic degradable polymer.

[0087] 49. The device of aspect 48, wherein the carrier is selected from PEG, PLA, PGA, PLGA, polymers thereof, and the like.

[0088] 50. The device of aspect 22, wherein the synthetic cellular receptor is selected from the group consisting of DDR1, DDR2, GPVI, LAIR-1, and OSCAR receptors.

[0089] 51. The device of aspect 22, wherein construct 3 binds to type III collagen via the synthetic cell receptor of aspect 50.

[0090] 52. The device according to aspect 22, wherein construct 3 binds to type III collagen via a collagen binding peptide present in vWF.

[0091] 53. The device according to aspect 52, wherein the collagen binding peptide is selected from

[0092] d. Any peptide containing the sequence GxRGQOGVMGFO.

[0093] e. Any peptide containing the sequence RGQOGVMGF.

[0094] f. Any peptide containing the sequence GPRGQOGVMGFO. BRIEF DESCRIPTION OF THE DRAWINGS

[0095] Embodiments of the present invention will be further described but are in no way limited by the following drawings.

[0096] Figure 1 is a list of amino acid functional groups commonly used as conjugation targets;

[0097] Figure 2 is a list of primary amine-reactive chemical groups;

[0098] Figure 3 It is a reaction scheme for the NHS ester-mediated coupling of carboxylic acids with primary amines of biomolecules;

[0099] Figure 4 It is a reaction scheme for coupling imidates with primary amines of biomolecules;

[0100] Figure 5 is a reaction scheme for carbodiimide-mediated coupling of carboxylic acids with primary amines of biomolecules;

[0101] Figure 6 is a scheme for the carbodiimide-mediated coupling of carboxylic acids with primary amines of biomolecules conjugated to sulfo-NHS;

[0102] Figure 7 is a reaction scheme for coupling maleimide to the thiol group of a biomolecule;

[0103] Figure 8 It is a reaction scheme for coupling iodoacetyl groups to thiol groups of biomolecules;

[0104] Fig. 9 is a reaction scheme for coupling pyridyl disulfide to thiol groups of biomolecules;

[0105] Fig. 10A The aryl ketone coupling mechanism is shown;

[0106] Fig. 10B is a reaction scheme for the coupling of aryl ketones with substrates having removable hydrogen;

[0107] Fig.11 Shown is the chemical structure of the reagent tetrakis(4-benzoylbenzyl ether) (TBBE) of pentaerythritol, which includes four potentially reactive benzophenone groups. DETAILED DESCRIPTION

[0108] Provided herein are collagen devices for treating cancer, methods for forming the collagen devices, and methods of using the collagen devices in treating cancer.

[0109] One aspect of the present invention relates to the digital processing of dental and medical devices and ECM scaffolds and cell scaffolds composed primarily of collagen, modified collagen and / or collagen-based peptides using 3D printing. In an exemplary embodiment, to produce polymerizable peptides, collagen is digested with an enzyme and then the peptides are modified with functional groups that can be polymerized using radiation. These modified peptides are then formulated with initiators, cross-linkers, solvents and / or other additives to create the desired design input for a specific dental or medical application. This formulation can then be 3D printed.

[0110] In another exemplary embodiment, collagen is digested to produce peptides, and other peptides are added or subtracted to produce custom desired design inputs for specific dental or medical applications (e.g., see Applicant's International Patent Application No. PCT / US2021 / 049646, published as WO 2022 / 060622A1, the disclosure of which is incorporated herein by reference). These newly formulated peptides are then modified with functional groups that can be polymerized with radiation. These modified peptides are then formulated with initiators, cross-linkers, solvents, and / or other additives to create the desired design inputs for specific dental or medical applications. This formulation can then be 3D printed. Another example involves extruding modified collagen to make it soluble in solvents and optionally modified with functional chemistry to allow for energy-driven post-processing. In an exemplary embodiment, the extrudable collagen may also contain other collagen-based peptides, such as collagen mimetic peptides ("CMPs", also known as collagen hybrid peptides ("CHPs"), to amplify the biological processes of collagen. In an exemplary embodiment, the extrudable collagen may also contain other bioactive peptides and growth factors, as well as cytokines, to enhance the healing process.

[0111] An exemplary procedure for producing a molded collagen-based dispersion or a reconstructed collagen matrix is ​​provided below. First, an acid dispersion of collagen fibers having a solid content of about 0.1 to 1.5% (w / w) is prepared. Both inorganic and organic acids or bases can be used. For example, a 0.05M to 0.1M collagen lactic acid dispersion having a pH of about 2.3 to 2.5 is prepared. An aliquot of insoluble collagen fibers is weighed and dispersed in an acid solution, homogenized using a commercial homogenizer, and filtered with a stainless steel mesh filter to obtain a collagen dispersion. The prepared collagen dispersion is then placed in a flask, and the pH value is adjusted to the isoelectric point of collagen (i.e., pH 5.0) by adding NH4OH for redissolution. The reconstructed collagen fibers are then taken out of the beaker and placed on a stainless steel sieve to remove excess solution until the desired solid content of the reconstructed collagen fibers is reached.

[0112] In another exemplary embodiment, the preparation of collagen dispersion and reconstruction of collagen fibers involves the use of alkali. Similar to the preparation of acidic dispersions, an alkaline dispersion of about 0.2-2.0% (w / w) of collagen fibers is dispersed in 0.001M-0.05M NaOH, homogenized, filtered to form a final collagen dispersion, and degassed under vacuum. The dispersion is then neutralized by adding HCl. The neutralized collagen dispersion is centrifuged and decanted to a certain amount of supernatant to achieve the desired density of the final reconstructed collagen mixture. The mixture can then be cross-linked and placed in a mold, then freeze-dried and finally sterilized.

[0113] The composition of the tumor extracellular matrix (ECM) is beginning to be recognized as playing a key role in the tumor microenvironment. During tumor growth, the ECM surrounding the tumor undergoes dramatic remodeling. The normal matrix is ​​degraded and replaced by an ECM with a higher collagen density and increased stiffness. The structure and density of this tumor-specific ECM supports tumor growth and metastasis and is therefore associated with a poor prognosis in several types of cancer, such as breast, pancreatic, and gastric cancer. This dense tumor ECM acts as a physical barrier, preventing the infiltration of immune cells, such as T cells and macrophages, which can attack and kill cancer cells. While collagen in healthy tissue acts as a network for the migration of activated T cells, dense collagen in the tumor microenvironment hinders immune cell infiltration and T cell migration (Boissonnas 2007). Furthermore, in addition to acting as a physical barrier, the collagen-dense ECM can reduce the proliferation of cytotoxic T cells, a T-cell subset that can directly kill malignant cells (Kuczek 2019), and direct tumor-associated macrophages to acquire an M2-like phenotype, which promotes tumor growth through immunosuppression and angiogenesis (Liu 2021). The dense ECM can also serve as a reservoir for immunomodulatory growth factors and act as a barrier to hinder the spread of chemotherapy.

[0114] Di Martino et al. (2021) demonstrated that cancer cells can secrete type III collagen into the extracellular space, inducing the cells to enter a dormant state. This is driven by the activation of discoidin domain receptor tyrosine kinase (DDR1), a receptor overexpressed in certain tumors that recognizes specific conserved sequences in collagen. The ECM surrounding these dormant cells is characterized by a loose, wavy collagen matrix that is partially enriched in DDR1-induced type III collagen, allowing immune cells (such as T cells) to invade and diffuse chemotherapy to tumor cells. Upon awakening of dormant cells, the extracellular domain of DDR1 is cleaved by MMPs. This domain of DDR1 is responsible for collagen fiber alignment, which contributes to the formation of a highly aligned and dense ECM, ultimately preventing T cell invasion and leading to failure of antitumor immunity. Monoclonal antibodies that recognize the extracellular domain of DDR1 can capture and prevent it from participating in collagen fiber alignment, allowing immune cell invasion (Sun 2021). Similarly, collagen-based peptides containing a conserved consensus sequence recognized by the DDR1 receptor, referred to herein as DDR1 peptides or DDR1-binding peptides, can capture the extracellular domain of DDR1 and prevent ECM reorganization. Alternatively, these peptides can also act as agonists of membrane-bound DDR1, inducing the expression of loose, wavy type III collagen that composes the ECM associated with tumor dormancy. T cells can also be genetically engineered or induced to express DDR1 peptides, promoting their migration and tumor infiltration in collagen matrices.

[0115] Recombinant human (rh) collagen is increasingly being used in medical devices and tissue engineering scaffolds due to concerns about the ability of animal-derived collagen to elicit an immune response in a small subset of the population. Recombinant human collagen has been developed in several stable or transient expression systems, including in yeast, mammalian cells, bacterial systems, and tobacco plants, as an alternative source of collagen with modifications that mimic the biological and mechanical functions of native collagen. This is particularly useful when the specific collagen to be isolated (such as type III collagen) is of low abundance, difficult to purify from donor tissue, or difficult to separate from other collagen types. The technology can also be used to produce recombinant human collagen-based peptides with sequence modifications designed to increase the density of cell-binding sites or to increase the uniformity of collagen fragments in a scaffold with desired specifications, such as increased DDR1 binding sites.

[0116] DDR1 has been shown to regulate collagen fiber alignment and induce type III collagen expression, ultimately controlling tumor cell dormancy. Various embodiments of engineered collagen devices that can regulate tumor cell dormancy are discussed below. In exemplary embodiments, such devices can be introduced locally into a patient at or near the tumor site by various methods, for example, in various embodiments, by surgical implantation, injection, or percutaneous placement (i.e., topical / percutaneous), or by non-surgical placement such as by a needle or catheter.

[0117] Membranes: Type III collagen membranes may be used in various exemplary embodiments. Degradation of the film over time should be optimized by controlling crosslinking, density, and porosity. The number of DDR1 binding sites in these devices can be increased by synthesizing DDR1 binding peptides and crosslinking the peptides into the device. Both native DDR1 binding peptides and added DDR binding peptides are released over time as the device degrades. The number of DDR1 binding sites (peptides) in these devices can also be increased by extracting isolated DDR1 peptides from digested type III collagen and crosslinking the peptides into the device. As the device degrades, both native DDR1 binding peptides as well as added peptides are released over time. DDR1 binding peptides can be added to the device (without crosslinking to the device) so that they can be released over time. The level of DDR1 peptides in the device can be reduced by mixing the device with various collagens, ECMs, GAGs, and other biocompatible polymers. One can also alter the DDR1 peptides in the type III collagen by genetically engineering rh type III collagen with varying amounts of DDR1 binding peptides.

[0118] As previously mentioned, the DDR1 receptor binds to and is activated by specific evolutionarily conserved sequences within collagen. The sequence GVMGFO, in which amino acid O is hydroxyproline, has been identified as a DDR1 binding motif in collagen. CHP and CMP are synthetic peptide sequences with Gly-Xaa-Yaa amino acid triplet repeating units (mimicking the signature sequence of collagen). These peptides can be designed and synthesized to contain a cell interaction domain and a collagen receptor binding motif, such as the DDR1 binding motif GVMGFO. These sequences are typically flanked by repeats of GPO or GfO, where f is 2S,4S fluoroproline, to maintain a triple helical hybridization tendency. Thus, these peptides can be in single, double, or triple helical form. Other binding sites and sequences that can bind and activate DDR1, as determined by collagen binding assays, cell membrane-bound DDR1 autophosphorylation, and activation of downstream signaling pathways, may also be incorporated. In various exemplary embodiments, these peptides can also be modified with positively or negatively charged amino acid side chains to control their diffusion through an electric field. Listed below are exemplary embodiments of potential DDR1 activating or DDR1 binding peptide sequences that can bind and / or activate DDR1 alone, or as previously described, can also be added or cross-linked to the device.

[0119] a. Any peptide containing the GVMGFO or GVMGFP sequence.

[0120] b. Any peptide containing the GPSGFO or GPSGFP sequence.

[0121] c. Any peptide containing the GPRGFO or GPRGFP sequence.

[0122] d. Any peptide containing the GARGFO or GARGFP sequence.

[0123] e. Any peptide containing the GQOGFO or GQOGFP sequence.

[0124] f. Any peptide containing the sequence GAMGFO or GAMGFP.

[0125] g. Any peptide containing the sequence in items 1-6, flanked by any number of GPO or GfO

[0126] Amino acid triplets, such as GPO(n)-GVMGFO-GPO(n) and GfO(n)-GVMGFO-GfO(n), where O is the amino acid hydroxyproline, f is 2S,4S fluoroproline, and n is the number of GPO or GfO repeats.

[0127] h. Any peptide sequence that can bind to DDR1, induce autophosphorylation of cell membrane-bound DDR1, or activate the DDR1 signaling pathway.

[0128] i. Any peptide from items 1 to 8 in single helical, double helical or triple helical form.

[0129] j. Any peptide in items 1-9 modified with positively or negatively charged amino acid side chains.

[0130] Layered membranes for cancer treatment:The type III collagen produced due to DDR1 activation helps to form a loose extracellular matrix and allows immune cells to invade and chemotherapy to spread, ultimately keeping tumor cells dormant and static. Various embodiments of an engineered type III collagen device that can be surgically implanted are discussed below. One such device comprises multiple layers (e.g., at least two layers) of type III collagen, at least some of which contain DDR1 binding peptide sequences. The first layer is cross-linked type III collagen, and the cross-linking density causes the layer to be absorbed / decomposed by digestion within approximately 6-12 months. The second layer is cross-linked type III collagen, so that the cross-linking density causes the layer to be absorbed / decomposed by digestion within approximately 3-6 months. The third layer is cross-linked type III collagen, and the cross-linking density causes the layer to be absorbed / decomposed by digestion within approximately 0-3 months. Due to size (thickness) and density, some layers may have more, less, or no cross-linking, but will still break at these time points. As this layered device decomposes over 12 months, it will release the DDR1 binding peptide from the collagen, enabling it to bind to the cleaved DDR1 receptors to prevent endogenous collagen fiber alignment and / or induce the production of endogenous type III collagen. The concentration of DDR1 peptide in each layer can be increased by chemically binding the DDR1 binding peptide to the device. As the device is digested, more and more DDR1 binding peptide will be released. The concentration of DDR1 binding peptide in each layer can be reduced by mixing type III collagen with other biomolecules or biocompatible polymers. As the device is digested / degraded, lower amounts of DDR1 binding peptide will be released. DDR1 binding peptides can be added to the device without binding, thereby releasing over time. In particular, the burst effect of DDR1 binding peptides at the time of device placement will be beneficial for immediate cancer prevention. Of course, cancer APIs can be added to the device for local treatment of cancer. In addition, modified DDR1 binding peptides bound to APIs can be added to the device. In this method, the API-modified DDR1 binding peptide will be released from the device and selectively bind to cancer cells, thereby delivering the API directly to cancer cells. Of course, the device can be designed with one or more layers and achieve similar performance by releasing the DDR1 binding peptide for a long time. Instead of using type III collagen as a carrier, biodegradable polymers with different degradation rates and loading the DDR1 binding peptide at different concentrations can produce devices that can inhibit cancer for extended periods of time. In various exemplary embodiments, the device can also be designed to release for more than 12 months. In other embodiments, the device can have less than or more than three layers. In various exemplary embodiments, these devices can be manufactured by additive manufacturing or molding.

[0131] Biodegradable film containing DDR1 peptide and type III collagen: In various exemplary embodiments, the above-mentioned films can also be engineered by utilizing biodegradable biomolecules or polymers as DDR1 peptides and type III collagen. For example, DDR1 peptides can be added to 3 layers, each layer being a different ratio of polylactic acid (PLA) and polyglycolic acid (PGA), resulting in different release rates of DDR1 peptides and type III collagen. In another example, 3 layers of hyaluronic acid containing DDR1 peptides or type III collagen, in which HA has different cross-linking densities, result in each layer degrading and releasing DDR1 peptides at 3 different rates. In various exemplary embodiments, DDR1 peptides or type III collagen can also be added to other collagen types, thereby producing various release curves, ultimately releasing DDR1 bound peptides.

[0132] Injectable device for treating cancer : In various exemplary embodiments, the device may also be an injectable device containing a DDR1 peptide. In various embodiments, the injectable device may include type III collagen, rh type III collagen (modified with DDR1 binding sites or native), DDR1 peptides and / or carriers, etc. These DDR1 binding peptides, whether bound to or released from the collagen matrix, can capture the extracellular domain of DDR1 and prevent it from participating in ECM reorganization. As previously described, these peptides can also activate membrane-bound DDR1, induce the expression and production of type III collagen, and thus promote ECM associated with tumor dormancy. The DDR1 peptide that binds to cancer cells can also have an API bound to the peptide as a means of selectively delivering the API to cancer cells. Another embodiment includes collecting T cells and genetically engineering or inducing them to express DDR1 peptides to promote their migration into the collagen matrix and tumor infiltration.

[0133] The overall approach of the injection device is to deliver DDR1 peptides to the cancer over an extended period of time. There are many ways to design such a device, including the following exemplary techniques:

[0134] 1) Type III collagen particles can be formed in various sizes, densities, and cross-linking concentrations, causing the particles to disintegrate at different rates, thereby releasing naturally occurring DDR1 peptides. For example, large, dense, and cross-linked type III collagen particles degrade faster than small, less dense, and cross-linked /

[0135] Non-crosslinked particles are slower. When type III collagen particles deliver DDR1 peptides to cancer cells, the particles can also serve as carriers. The particles can be loaded with additional DDR1 binding peptides to increase the overall DDR1 peptide concentration.

[0136] 2) Many other carriers, including particulate forms, can be used to deliver DDR1 peptides to the cancer cells for a long time: type III collagen, other collagen types, glycosaminoglycans (GAGs), PLA, PGA, polylactide-co-glycolic acid (PLGA), polyurethane (PU) and other biodegradable / bioresorbable polymers, hydrogels and biomolecules.

[0137] 3) The above particles can be suspended in various biocompatible aqueous solutions, including water, buffer, saline that may contain other osmotic agents. In most cases, the suspension of the particles will occur before injection. Of course, the aqueous suspension can also contain DDR1 peptides, which also include digested type III collagen fragments.

[0138] 4) The above particles used in the injection device can also be added to the above film.

[0139] 5) Much like the DDR1 peptides conjugated to API used in membrane devices, this molecule / peptide can be used in an injectable device.

[0140] 6) In the above injection example, genetically engineered collagen to which DDR1 peptide has been added can be used.

[0141] Disposable device for treating cancer : In various exemplary embodiments, the devices described in the above sections of the injectable devices and films for cancer treatment can also be used topically (i.e., transdermally / percutaneously) to treat cancer. Other local devices can be used to deliver the DDR1 peptide to the cancer site. For example, type III collagen can be processed into a soluble gelatin form and used topically. Type III gelatin can also contain additional DDR1 peptides to change the concentration. The DDR1 peptide can be added to a number of local carrier formulations to deliver the peptide to the treatment site. Many of these formulations are well known in the art, including, for example, micelles, lipid nanoparticles, lipid carriers, microemulsions, and liposomes. In another embodiment, a DDR1 peptide bound to an API described in a membrane device, the molecule / peptide can be used in a local device.

[0142] Other methods to alter DDR1 peptide concentration in collagen / biomolecules : Type III collagen contains DDR1 peptides that bind to DDR1 binding sites. In an exemplary embodiment, rh type III collagen can be engineered to contain an increased amount of DDR1 binding sites. This approach is another way to control the concentration of DDR binding peptides in the above-mentioned device. In fact, using this technology, DDR1 binding sites can be incorporated into any protein or any type of collagen. DDR1 peptides themselves can also be produced in this way. Many proteins or biomolecules can be genetically engineered to include DDR1 peptide sequences. New proteins or biomolecules can be designed similar to what is disclosed in this application.

[0143] In various exemplary embodiments, cancer-forming T cells can be genetically engineered or induced to express DDR1 peptides, promoting their migration and tumor infiltration in collagen matrices.

[0144] The modified cells can be integrated back into the patient to treat cancer. Prior to using the CART-T method, the patient is preferably treated with a device of the present disclosure.

[0145] The liver and pancreas contain highly enzymatic environments. This can cause rapid degradation of protein-based devices. To address these environments, the aforementioned films can contain a layer of biodegradable polymers (via a hydrolysis process) to help act as a scaffold or possible DDR1 peptide library. The aforementioned biodegradable polymers can also be used in this approach.

[0146] Although in other exemplary embodiments, the use of type III collagen is described herein as a way to deliver DDR1 peptides to cancer in a patient. Collagen can also serve as a scaffold for tissue regeneration. Many of the devices described herein are also used as tissue regeneration devices.

[0147] Although type III collagen contains a DDR1 binding site, one can genetically engineer most proteins or biomolecules to contain DDR1 binding sites and design similar devices using these novel biomolecules. Although type III collagen has a typical DDR1 binding motif that can activate DDR1, other types of collagen, such as type II and type IV collagen, also have this DDR1-activating DDR1 binding motif or can be engineered to have such a binding site. Other types of collagen may have sequences that have a weaker affinity for DDR1 and can also activate DDR1. Therefore, they can also be used in the above devices.

[0148] DDR peptides enhance immune responses to co-administered therapeutic agents to treat cancer : The innate immune system routinely eradicates cancer cells (Pandya et al., 2016), but sometimes such cells evade immune cells and form tumors. As Gonzalez et al. (2018) assert, "In principle, tumor development can be controlled by cytotoxic innate and adaptive immune cells; however, as tumors progress from neoplastic tissue to clinically detectable tumors, cancer cells evolve different mechanisms to mimic peripheral immune tolerance to avoid tumor invasion."

[0149] Although the treatment of cancer in patients by delivering DDR1 peptides, or using implanted collagen devices to deliver DDR1 peptides, is new, unrelated drug-based and emerging cancer therapies are known and routinely practiced. These drugs include cancer APIs designed to kill or inhibit tumor cell metabolism and proliferation, such as bleomycin, capecitabine and dacarbazine, doxorubicin, eribulin, gemcitabine, ixabepilone, lapatinib paclitaxel, and vinblastine sulfate (Cortazar et al., 2012), as well as novel therapeutic approaches designed to elicit immune responses against tumor-specific antigens and prevent tumor cells from inactivating T cells that destroy them, such as cancer vaccines (https: / / www.cancer.gov / about-cancer / treatment / drugs / abd). In addition, new therapeutic drugs are being discovered every year. For example, U.S. Patent Publication No. 20230022524 (the entire contents of which are incorporated herein by reference) discloses heterobifunctional compounds as inhibitors and degraders of hematopoietic progenitor kinase 1 (HPK1) (You et al., 2021; Si et al., 2020), which is a negative regulator of T cells (Sawasdikosol et al., 2020).

[0150] As explained by NIH's National Cancer Institute, "Immune checkpoints are a normal part of the immune system. They work by preventing immune responses from becoming too intense and destroying healthy cells in the body. Immune checkpoints engage when proteins on the surface of immune cells called T cells recognize and bind to partner proteins on other cells, such as some tumor cells. These proteins are called immune checkpoint proteins. When the checkpoint and partner proteins bind together, they send an 'off' signal to the T cell. This stops the immune system from destroying the cancer."

[0151] Immunotherapy drugs called immune checkpoint inhibitors work by blocking the binding of checkpoint proteins to their partner proteins. This prevents the "off" signal from being sent, allowing T cells to kill cancer cells. One of these drugs acts on a protein called CTLA-4. Other immune checkpoint inhibitors act on a checkpoint protein called PD-1 or its partner protein PD-L1. Some tumors reduce T cell responses by producing large amounts of PD-L1 (https: / / www.cancer.gov / about-cancer / treatment / types / immunotherapy / checkpoint -inhibitors ).”

[0152] The development of therapeutic (rather than preventive) vaccines has accelerated over the past two decades. Therapeutic cancer vaccines are generally designed to induce immune responses that prime endogenous tumor-reactive T cells against tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs) and prevent T cells from being inactivated by cancer cells (Lin et al., 2022). Clinical results of cancer therapeutic vaccines have been mixed, ranging from frustrating failures to great successes.

[0153] Examples of immunotherapeutic vaccines described in the literature include glycoprotein 100 (gp100) vaccine (Tahaghoghi-Hajghorbani et al., 2023), Epstein-Barr virus (EBV) target antigen vaccine (Taylor et al., 2014), synthetic long peptide (SLP) vaccine ISA101 (Ding et al., 2022), E6 / E7 plasmid (VGX-3100), and E6 / E7 / Fms-like tyrosine kinase 3 ligand (Flt3L)-plasmid (GX-188E) vaccine (Kim et al., 2014), E6 / E7IL-2MVA vector vaccine (Harper et al., 2019), CDX-1401 and Resci model (TLR7 / 8) and Hiltonol (polyICLC, TLR3) vaccines (Dhodapkar et al., 2014) and adenovirus (ChAdOx1) / MVA vaccine targeting MAGE-A3 and NY-ESO-1 (McAuliffe et al., 2021).

[0154] Examples of immunotherapeutic vaccines described in the literature that have failed to achieve meaningful clinical efficacy include the Rindopepimut (CDX-110) vaccine (Weller et al., 2017), the TLR4 agonist adjuvant (AS02B) MAGE-A3 protein vaccine (Vansteenkiste et al., 2013), the Nelippimut-S plus GM-CSF vaccine (Mittendorf et al., 2019), and the single-epitope HLA-II-restricted 15-mer peptide (AE37) + GM-CSF vaccine (Mitendorf et al., 2016).

[0155] Examples of commercially available clinical immunotherapy vaccines include atezolizumab (Tecentriq) (https: / / www.tecentriq.com), avelumab (Bavencio) (https: / / www.bavencio.com / hcp), cemiplizumab (Libtayo) (https: / / www.libtayo.com), durvalumab (Imfinzi) (https: / / www.imfinzi.com), enzalutamide (XTANDI) (https: / / www.xtandi.com), ipilimumab (Yervoy) (https: / / www.yervoy.com) , Opdivo (https: / / www.opdivo.com), Opdualag (https: / / www.opdualag.com), Keytruda (https: / / www.keytruda.com), Rituxan (https: / / www.rituxan.com), and Imjudo (https: / / www.accessdata.fda.gov / drugsatfda_docs / label / 2022 / 761289lbl.pdf). .

[0156] Many other cancer vaccines based on shared antigens, anonymous antigens, personalized antigens, autologous tumor lysates, and oncolytic viruses and bacteria administered intratumorally are currently in development or being evaluated in registered clinical trials. For example, Moderna (Cambridge, MA) publicly lists three cancer pipeline vaccines on its website, namely personalized cancer vaccine (PCV) mRNA-4157, KRAS vaccine mRNA-5671, and checkpoint vaccine mRNA-4359 (https: / / www.modernatx.com / research / product-pipeline). Similarly, other drug manufacturers have reported that cancer vaccines are in development.

[0157] Lin et al. (2022) concluded, "The data suggest that inducing T cells against self-proteins, even those overexpressed in tumors, requires an enhanced immune response to achieve maximal efficacy." In another aspect of the invention, the combined use of DDR1 peptides and any anti-cancer therapeutic agent enhances the efficacy of the therapeutic agent by enhancing an effective immune response. Engineered type III collagen devices (including films, particles, and other carriers) containing DDR1 binding peptides are delivered or implanted as an adjunct to drug or immune cancer therapy to prevent the formation of dense tumor ECM, which acts as a physical barrier to block the infiltration of immune cells (such as T cells and macrophages) that can attack and kill cancer cells, while the therapeutic agent directly attacks the tumor or induces an immune response against the tumor and promotes T cell attack on the tumor. The combined use of DDR1 peptides and checkpoint inhibitors or DDR1 peptides with other cancer vaccines is particularly attractive because each vaccine promotes an increase in T cell activity and the combination enhances the effective reactivity of T cells against tumor cells. Furthermore, simultaneous use of DDR1 peptides could render cancer vaccines unsuccessful, as the combined effects of DDR1 peptides and vaccines could achieve T cell activity that exceeds the threshold required for effective tumor eradication.

[0158] A preferred example is a combination of a DDR1 peptide that promotes type III collagen formation and inhibits the formation of a physical barrier to prevent immune cell-mediated tumor cell death and a therapeutic agent described in U.S. Patent Publication No. 20230022524 (incorporated by reference in its entirety) that inhibits and degrades HPK1, which otherwise mediates T cell escape of tumors. Two embodiments of the present invention are particularly suitable, the first is an HPK1 inhibitor / degrader and an accompanying engineered type III collagen membrane, which contains a DDR1 binding peptide sequence flanked by repeating sequences of GFO or GPO, wherein F is fluoroproline and O is hydroxyproline, i.e., (GPO)6 on each side, to maintain the triple helical form of the peptide at physiological temperature, and the second is an accompanying particle comprising one or more layers of a degradable polymer containing the DDR1 peptide, which delivers the peptide at a controlled rate when the polymer degrades.

[0159] DDR1-targeting peptides for local delivery of therapeutic agents to tumors: In various exemplary embodiments of the invention, the same DDR1 peptide that promotes type III collagen formation can be attached with an anti-cancer therapeutic (including cancer APIs designed to kill tumor cells and vaccines designed to elicit an immune response against tumors) connected by a degradable linker to form a peptide-drug conjugate. Chavda et al. (2022) described the general concept of using peptides as drug transporters in this manner. They defined a peptide drug conjugate as consisting of: 1. a homing peptide or device; 2. a cytotoxic payload; and 3. a synergistic linker that delivers the cytotoxin to a target receptor on the cancer cell. The present invention includes constructs in which the DDR1 peptide serves as a homing peptide, but alternatively includes an immunotherapy rather than a cytotoxin as a payload. U.S. Patent Publication No. 20210338558 (incorporated herein by reference in its entirety), claim 1 describes a similar construction consisting of: 1. a targeting portion for binding to native collagen fibers; 2. a skin care agent or cosmeceutical payload; and 3. an intermediate release linker bound to the skin care agent or cosmeceutical; the inventors recognized that the peptide GVMGFO is a DDR1 targeting portion.

[0160] Suitable linkers for peptide-drug conjugates depend on the specific peptide / drug combination. Examples of drug linker-peptide combinations include gemcitabine glutaryl linker-[D-Lys6]-gonadotropin-releasing hormone (GnRH), paclitaxel ester linker Angiopep-2, paclitaxel ester linker-Polishrink, thapsigargin linker Tetrapide, doxorubicin ester linker GnRH, maytansine disulfide linker bicyclic peptide, doxorubicin amide linker Tetrapide, 111In-DTPA-amide linker-D-phe-1-octreotide, hTNF-amide linker-NGR, paclitaxel ester linker-Poliwrink, CLIP71-amino linker-LHRH, gemcitabine amide, ester, cathepsin B, carbamate linker knot peptide, DOTA-g-aminobutyric acid linker bombesin 7-14, methotrexate amino linker acetyl YTAIAWVKAFIRKLRK (Chavda et al., 2022). U.S. Patent Publication No. 20210338558 also discloses various intermediate release linkers; these include proteins or peptides, non-protein polymers such as polyethylene glycol (PEG), PEG-modified proteins and biotin / avidin, biotin / streptavidin, antigen / antibody or hapten / antibody linkages. Examples of peptide linkers include TGEKP, TGGGGSGGGGTGEKP, modifications of TGGGGSGGGGTGEKP, LRQKDGGGSERP, LRQKDGERP, GGRGRGRQ, QNKKGGSGDGKKKQHI, TGGERP, ATGEKP, GGGSGGGGEGP, and derivatives of those linkers in which amino acid substitutions are made, such as in QNKKGGSGDGKKKQHI or GGGSGGGGEGP, the serine (S) residue between the diglycine or polyglycine sequence is replaced by threonine (T), the glutamic acid (E) at position 9 in GGGSGGGGEGP is replaced by aspartic acid (D), and other linkers such as glycine or serine repeats. When both the therapeutic agent and the intermediate release linker are peptide-derivatized, the connection between the therapeutic agent and the intermediate release linker can be a peptide (amide) bond formed between these peptides. Additional cleavable linkers include disulfide-containing linkers, which are cleaved by reduction; diol-containing linkers, which are cleaved by periodate; diazo-containing linkers, which are cleaved by dithionate; ester-containing linkers, which are cleaved by hydroxylamine; and sulfone-containing linkers, which are cleaved by base.

[0161] Typically, the connection between the anticancer therapeutic agent and the intermediate release linker and between the intermediate release linker and the targeting peptide is a covalent connection involving reactive moieties and cross-linking agents known in the art. In addition, as understood in the art, the connection can be covalent, through interactions such as hydrophobic / hydrophilic interactions, hydrogen bonds, ionic interactions or salt connections.

[0162] Peptide linkers are particularly attractive because they can be seamlessly attached to the synthetic DDR1 peptides of the present invention using a standard peptide synthesizer. Optionally, the peptide linker can be substituted with polyethylene glycol (PEGylation) to infuse it with desired physical and chemical properties to aid in the delivery of the attached drug. Once formed, the [(DDR1)-(linker peptide)] construct can be attached to the therapeutic agent by non-covalent means as described above, but more typically by covalent bonding, using wet chemistry and photochemical techniques known in the art.

[0163] In some embodiments, the linker peptide may be modified with one or more heat or photoreactive groups capable of covalently linking to the therapeutic agent (or DDR1 peptide, if not already synthetically linked). In other embodiments, the therapeutic agent may be modified with one or more heat or photoreactive groups capable of covalently linking to the linker peptide (or [(linker peptide)-(DDR1 peptide)] conjugate.

[0164] The linker peptide (or [(linker peptide)-(DDR1 peptide)] conjugate) can be linked to the therapeutic agent, and the choice of method depends on the chemical structure of the therapeutic agent. Cross-linking agents that react with various reactive chemical moieties, including hydroxyl (-OH), primary amine (-NH2), sulfhydryl (-SH), and carboxyl (-COOH), are known in the art. The therapeutic agent can be easily modified with the linker peptide of the present invention by first reacting the cross-linker with the reagent and then with the linker peptide, or vice versa. This can be achieved by various methods, including wet chemical and photochemical methods.

[0165] Wet chemistry methods useful in the present invention are described in the Thermo Scientific Crosslinking Technology Handbook (Thermo Fisher Scientific Inc.: Waltham, MA, 2012, available from https: / / tools.thermofisher.com / content / sfs / brochures / 1602163-Crosslinking-Reagents-Handbook.pdf. Accessed on July 28, 22, incorporated herein by reference). Figure 1 Chemical moieties that are often targeted for bioconjugation are shown; Table 1 lists common chemical cross-linkers that react with these moieties. With respect to the present invention, many anti-cancer vaccines are protein-based vaccines, which makes them attractive for bioconjugation. In contrast, many APIs, while not protein-based, are reactive with the same chemical cross-linkers used for bioconjugation.

[0166] Table 1

[0167] Commonly used cross-linker reactive groups for protein coupling (Table 1 in Thermo Scientific Cross-linking Technology Manual)

[0168]

[0169]

[0170] Amine-reactive chemical groups target primary amines at the N-terminus of polypeptide chains (alpha amines) as well as side chains of lysine residues (epsilon amines). Primary amines are often located on the surface of proteins due to their positive charge under physiological conditions, thus allowing bioconjugation without denaturation. Figure 2 Chemical groups that react with primary amines are shown; the most commonly used are NHS esters and imido esters. Figure 3 and 4 The general cross-linking reaction schemes for these reagents are shown separately.

[0171] Carboxylic acid reactive chemical groups target the carboxyl C-terminus of polypeptide chains, as well as the side chains of aspartic acid and glutamic acid. Carboxyl groups are also often located on the surface of proteins and can therefore be used for bioconjugation.

[0172] Carbodiimides (EDC and DCC) result in direct conjugation of carboxylates (-COOH) with primary amines (-NH2). Figure 5 Shown is a general reaction scheme for the EDC-mediated coupling of a carboxylic acid of one biomolecule (eg, a linker peptide) to a primary amine of another biochemical molecule (eg, a therapeutic agent). Figure 6 A general reaction scheme for EDC coupling with sulfo-NHS conjugation is shown, which improves the reaction efficiency and produces a stable sulfo-NNS-derivatized biomolecule (peptide or therapeutic agent) that can be stored and used later after reaction with the primary amine of a second complementary biomolecule (therapeutic agent or peptide).

[0173] Sulfhydryl-reactive chemical groups target the side chains of cysteine ​​residues. Sometimes disulfide bonds (-SS-) form between adjacent polypeptide side chains in proteins, which must first be reduced to sulfhydryl groups before cross-linking via these groups. Maleimide, haloacetyl, and pyridyl disulfide moieties all react with protein sulfhydryls. Figure 7-9 A general reaction scheme is shown for the coupling of maleimide, haloacetyl, and pyridyl disulfide groups to biomolecule sulfhydryl groups, respectively. Maleimide chemistry is often used in conjunction with NHS ester chemistry to produce heterobifunctional crosslinkers that enable controlled two-step coupling of peptides to therapeutic agents.

[0174] In addition to amino acids with amino, carboxyl or thiol side groups, in various exemplary embodiments, other chemicals with amino, carboxylic acid or thiol groups can also be used to cap the connecting peptide. Preferably, such chemicals have a terminal amino or carboxyl group that can react with the C-terminal carboxyl or N-terminal amino group of the connecting peptide, and multiple amino, carboxyl or thiol side groups, or a combination thereof, to target bioconjugation to the therapeutic agent with higher efficiency than unmodified peptides.

[0175] Polyamine-containing chemicals include propane-1,2,3-triamine (three NH2 groups), tris(2-aminoethyl)amine (four NH2 groups), 1,1,2,2-3-ethylenediaminetetraacetic acid (four NH2 groups) and 2,2-bis(aminomethyl)propane-1,3-diamine (four NH2 groups).

[0176] Polycarboxylic acid-containing chemicals include propane-1,2,3-tricarboxylic acid (three COOH groups) and citric acid (three -COOH groups).

[0177] Polymercapto chemicals include 2,3-dimercaptopropionic acid (one terminal COOH and one terminal SH group, as well as one additional SH group) and 2,4-dimercaptoglutaric acid (two terminal COOH groups and two SH groups)

[0178] Chemicals with multiple amino and carboxyl groups include: 2,2-diaminoacetic acid (one terminal COOH and one terminal NH2 group, plus one NH2 group), 2,4-diaminobutyric acid (one terminal COOH and another terminal NH2 group, plus one NH2 group), 2,3-diaminopropionic acid (one terminal COOH and another NH2 group), 2,4-diaminoglutaric acid (two terminal COOH groups, two other NH2 groups.

[0179] In addition to wet chemical methods, photochemical methods can also be used for bioconjugation. US Patent No. 5,563,056 (incorporated herein by reference) describes potential reactive groups and activated residual functional groups, as shown in Table 2.

[0180] Table 2 Potential reactive groups and their activated residues described in U.S. Patent No. 5,563,056

[0181]

[0182]

[0183] Fig. 10A and 10BA general photocoupling reaction of aryl ketones using derivatized benzophenone as an example is shown (see Simso EJ. PhotoLink: a new coating concept, Textile Technology International. 1996; 1:23, available at https: / / p2infohouse.org / ref / 33 / 32073.pdf). U.S. Patent No. 5,744,515 describes (incorporated herein by reference) the related coupling mechanism as follows. "Photoreactive aryl ketones, such as acetophenone and benzophenone, or derivatives thereof, are preferred because these functional groups generally readily undergo the activation / deactivation / reactivation cycles described herein. Benzophenones are a particularly preferred photoreactive group because they are capable of photochemical excitation, initially forming an excited singlet state, which undergoes intersystem crossing into a triplet state. The excited triplet state can then insert into a carbon-hydrogen bond by abstracting a hydrogen atom (e.g., from the support surface), thereby forming a free radical pair. Subsequent collapse of the free radical pair results in the formation of a new carbon-carbon bond. If the UV-induced excitation of the reactive bond (e.g., carbon-hydrogen) of the benzophenone group is reversible, the molecule returns to the ground state energy level upon removal of the energy source. Photoactivatable aryl ketones, such as benzophenone and acetophenone, are particularly important because these groups will reactivate multiple times in water, thereby increasing coating efficiency. Therefore, photoreactive aryl ketones are particularly preferred."

[0184] Particularly useful in this regard is a heterobifunctional crosslinker having a photoreactive benzophenone group at one end and an amine-reactive N-oxysuccinimide (NOS) ester group at the other end, the two ends being linked together by an ε-aminocaproic acid (EAC) spacer. This compound can be used to attach the NOS terminal to proteins and peptides via their primary amines, thereby producing a photoderivatized peptide or protein that can be bioconjugated to any material capable of free radical reaction with excited benzophenone. U.S. Patent Nos. 5,744,515 and 6,121,027 describe examples of photoreactive proteins and peptides, including fibronectin, laminin, and type IV collagen, and peptides of fibronectin (RGD, C / HV, C / H-II), laminin (F-9), and type IV collagen (HEP-III), respectively. The chemical methods described in U.S. Patent No. 6,121,027 are particularly suitable for preparing the photoreactive peptides of the present invention, and the patent is incorporated herein by reference.

[0185] In the case of a photoderivatized protein-based therapeutic, the NOS terminus of the cross-linker is bound to the therapeutic molecule via its primary amine, while the benzophenone terminus remains free to bind to the linker peptide of the invention in close contact and upon subsequent illumination. In the case of a photoderivatized linker peptide, the benzophenone terminus of the cross-linker remains free to bind to the therapeutic molecule at any CH group of the amino acids that compose it, some of which are more reactive than others.

[0186] Other photocrosslinkers can also be used to bind the connecting peptides of the invention to the therapeutic agents of the invention. As first disclosed in U.S. Pat. No. 5,414,075, a constrained multifunctional reagent containing multiple potentially reactive groups for surface modification can first bind to a support structure after activation and then bind to a different second molecule after reactivation. For example, Fig.11 The chemical structure of the reagent tetrakis (4-benzoylbenzyl ether) of pentaerythritol (TBBE) is shown, which includes four potentially reactive benzophenone groups. These multifunctional reagents are easily combined first with the therapeutic agent of the present invention and then with the connecting peptide of the present invention. In another embodiment, the [(therapeutic agent)-(connecting peptide)-(DDR1 peptide)] construct, referred to herein as Construct 1, can be formulated with a biodegradable carrier so that it can be released over time.

[0187] In other embodiments, the [(therapeutic agent)-(connector peptide)-(DDR1 peptide)] construct is added to the reconstituted (or otherwise modified) collagen such that it affects the pharmacokinetic or release properties of the former.

[0188] Collagen targeting peptides other than DDR1 for local delivery of therapeutic agents to tumors: In various exemplary embodiments of the present invention, anticancer therapeutic agents are delivered to tumors by utilizing additional ligand-non-DDR1 receptor interactions. This is achieved by using one of two different constructs. In the first construct, the above-mentioned [(therapeutic agent)-(connecting peptide)] is bound to a ligand that binds to a collagen receptor on a tumor cell, including a DDR2 receptor, select integrin receptors (α1β1, α2β1, β10β1, and α11β1), select immunoglobulin (IgG)-like receptors, glycoprotein VI (GPVI), leukocyte-associated immunoglobulin-like receptor 1 (LAIR-1), osteoclast-associated receptor (OSCAR), and mannose receptor (Agarwal et al., 2019). The choice of ligand depends on the type of tumor cell and its collagen receptor. Such ligands are typically peptides, but can also be other structures. The conjugate is similar to the conjugate of the previous aspect, but the DDR1 peptide is replaced by a peptide that binds to a non-DDR1 receptor [(therapeutic agent)-(connecting peptide)-(non-DDR1 peptide)], referred to herein as construct 2. Targeting ligands that bind to non-collagen receptors can also be used. In the second construction, the therapeutic agent linker peptide is linked to a synthetic receptor or targeting peptide that binds a ligand in type III collagen, and the [(therapeutic agent)-(linker peptide)-(type III collagen receptor or targeting peptide)] conjugate bound to a type III collagen device (particle or film) (referred to herein as construct 3) is placed at or near the tumor site. Alternatively, the therapeutic agent linker peptide is linked to a synthetic receptor or targeting peptide that binds a ligand in type I collagen, such as a DDR or integrin receptor, to produce a [(therapeutic agent)-(linker peptide)-(type I collagen receptor)] construct, referred to herein as construct 4 (particle or film), and delivered to the tumor to take advantage of the overexpression of type I collagen in many tumors (Kanta 2015; Hsu et al. 2022).

[0189] GVMGFO and some of the additional peptide ligands for DDR1 listed above are also ligands for DDR2, and in addition to GVMGFO identified using the ColII and ColIII toolkit peptides, there are additional DDR2-binding peptides (Agarwal G et al., 2019); some ligands are common to both DDR1 and DDR2, while others are specific ligands for either DDR1 or DDR2 (Leitinger et al., 2014). The DDR2-binding peptides shown in Table 2 (Konitsiotis et al., 2008, Table 1) and Table 3 (Xu et al., 2011, Table 3) can be easily linked to a linker peptide in a single synthetic step, and the appropriate terminal amino acid can be attached for chemical coupling to a therapeutic agent.

[0190] Similar to targeting peptides, in various exemplary embodiments, targeting antibodies can also be used to deliver therapeutic agents to tumor sites. For example, DDR monoclonal antibodies are described in the academic literature (Carafoli F et al., 2013). Like [(therapeutic agent)-(connecting peptide)-(DDR1-peptide) and [(therapeutic agent-connecting peptide-DDR2-peptide)] constructs, [(therapeutic agent)(connecting peptide)(DDR monoclonal antibody)] constructs target DDR receptors in vivo.

[0191] Table 2

[0192] DDR2 binding peptide

[0193] (Table 1 of Konitsiotis et al.)

[0194] Table 1 Guest sequences of peptides used to characterize DDR-2-collagen interactions 1

[0195]

[0196] 1 DDR2 binding indicates binding to His-DDR2 or DS2-Fc. Plus sign (+) indicates similar response to peptide II-22, II-23 or GDS-GFO (control); minus sign (-) indicates less than 20% of the control group; plus and minus sign (+ / -) indicates between 20% and 80% of the control group.

[0197] 2 GAP - GPR-GFO: non-helical peptide flanked by (GAP) 5 instead of (GPP) 5 like all other peptides.

[0198] Table 3

[0199] DDR1 and DDR2 binding peptides

[0200] (Table 1 of Xu et al.)

[0201] Table 1 Guest sequences of peptides used to characterize DDR collagen interactions a

[0202]

[0203] a DDR1 binding indicates binding to DDR1-Fc or DDR1-Fc2; DDR2 binding indicates binding to DDR2-Fc or His-DDR2. Plus sign (+) indicates similar reaction to peptide II-22, III-23, III-23 or GPS-GFO (control); minus sign (-) indicates less than 10% of control; (+) / - indicates 10-30% of control; + / - indicates 30-80% of control

[0204] b The data include those of Konitsiotis et al. (2008) J. Biol. Chem. 283, 6861-6868.

[0205] c GAP - GPR-GFO: non-helical peptide flanked by (GAP) 5 instead of (GPP) 5 like all other peptides.

[0206] In various exemplary embodiments, the [(therapeutic agent connecting peptide-integrin targeting peptide)] construct can also be used to deliver anti-cancer therapeutics using the integrin receptors of different collagens. For example, the collagen binding A domains α1β1 and α2β1 of integrins recognize the GFOGER sequence in native type I and type II collagen (Knight et al., 2000). Farndale (2019) summarizes various integrin collagen peptide sequences that can be used in the present invention. The GxOGER sequence appears in type I, II, and III collagens, while GROGER appears in type I and III collagens, but not in type II collagen. Different collagen sequences selectively bind to different integrins, α1 and α10β1 have affinity for GLOGEN in type III collagen, while α2 and α11β1 have higher affinity for GFOGER in type I, II, and IV collagens, etc. Integrin α2β1 binds to several GXX'GER motifs in collagen, while the GXX'GEX' motif supports binding of all collagen-binding integrins (Farndale et al., 2008). The GLPGER and GFPGER sequences in streptococcal collagen-like proteins bind to both the α1 and α2I-domains without the need for hydroxyproline, while GFPGEN binds only to the α1I-domain (Seo et al., 2010).

[0207] IgG-like receptors bind collagen via distinct motifs. GpVI has a more complex recognition of collagen than α2β1; the GPO triad is essential but not sufficient for binding (Farndale et al., 2008). The binding peptides in the collagen toolkit all contain OGPOGP, and two AGPOGP motifs also contribute to GPVI binding (Feitsma et al., 2022). GPVI is a platelet membrane protein consisting of 319 amino acid residues with a molecular weight of 62 kDa (Jung et al., 2008). The structure of the collagen binding domain (CBD) of GPVI is known, and recombinant GPVI-CBD has been prepared; similar to related immune receptors, GPVI contains two vertically arranged immunoglobulin-like domains (Horii et al., 2006). LAIR-1, like GpVI, recognizes the GPO triplets found in type I and type III collagens, but sequences containing more complex GPOs in collagen provide their natural ligands (Farndale et al., 2008). LAIR-1, also known as CD305, is a type I glycoprotein composed of 287 amino acids that was first molecularly cloned in 1997 (Van Laetem et al., 2022). Its structure contains an extracellular C2-type Ig-like domain and two immunoreceptor tyrosine-based inhibitory motif (ITIM) domains in the cytoplasmic tail. The minimal OSCAR-binding collagen sequence identified using hOSCAR Fc fusion protein is GPOGPX'GFX' (Barrow et al., 2011). One such binding peptide is triple helix (GPO)3GPOGPAGFO(GPO)2G, as the triple helical structure of collagen is essential for OSCAR binding. The three-dimensional structure of the extracellular region of human OSCAR consists of two Ig-like domains, D1 (membrane distal) and D2 (membrane proximal), connected by a short interdomain linker (Haywood et al., 2016).

[0208] In various exemplary embodiments, other targeting peptides may also be used in the [(therapeutic agent-linker peptide-targeting peptide)] construct, depending on the specific tumor cell type and its respective target. For example, MMP-1 is overexpressed in triple-negative breast cancer (Wang et al., 2019), and the GPQG~LAGQRGIVGLOGQRGER and GPOGPQG~LAGQRGIVGLOGQRGER motifs bind to MMP-1 (Farndale 2019). Similarly, MMP-13 is overexpressed in various cancers and plays multiple roles in tumor progression and metastasis (Li et al., 2022), and GPQG~LAGQRGIVGLOGQRGER binds to MMP-13 (Farndale 2019). Integrin α5β1 is overexpressed in many tumors (Hou et al., 2020), and the novel peptide of the RX1ETX2WX3 sequence described in U.S. Patent No. 7,067,619 binds to α5β1-integrin (incorporated herein by reference in its entirety).

[0209] While therapeutic agents can be delivered to the tumor site via a linker peptide that directly binds to a cell receptor on the cancer cell, in various exemplary embodiments, this can also be achieved by linking a synthetic cell receptor that binds to a peptide motif in a type III collagen device. Although type III collagen devices are preferred based on their ability to promote loose collagen structure, the [(therapeutic agent)-(linker peptide)-(synthetic cell receptor)] conjugate referred to herein as construct 5 can also be bound to devices made from other collagen types (such as type I collagen), with the specific receptor depending on the specific collagen type. The device can be in the form of particles that are injected directly into or near the tumor, or first encapsulated in a degradable carrier prior to injection. In various exemplary embodiments, the device can also be in the form of an inlay or film that is surgically implanted at or near the tumor site.

[0210] Von Willebrand factor (vWF) binds to type III collagen (Romijn et al., 2003) via its A3 domain at a single high affinity site, GxRGQOGVMGFO (Farndale 2019), where RGQOGVMG is the minimal binding sequence (Lisman et al., 2006). Interestingly, GPRGQOGVMGFO in type III collagen also binds to the DDR2 receptor (Farndale et al., 2008). Thus, the [(therapeutic agent)-(linker peptide)-GPRQOGVMGFO) conjugate, referred to herein as construct 6, could be placed directly at or near the tumor site, or linked to type III collagen that is subsequently placed at or near the tumor, or both. “Throughout all of this, the amino acid sequence of the collagen molecule contains a myriad of potential ECM ligand interaction sites, including the DDR, that are aligned within the D phase separated by microns (laterally and axially) (Orgel et al., 2019)”, suggesting the use of other, yet-to-be-discovered targeting peptides.

[0211] All features disclosed in this specification may be combined in any combination.

[0212] Each feature disclosed in this specification can be replaced by an alternative feature used for the same, equivalent or similar purpose. Therefore, unless otherwise explicitly stated, each feature disclosed is only an example of a series of equivalent or similar features.

[0213] From the above description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

[0214] In general, any combination of the disclosed features, components and methods described herein is possible. The steps of a method may be performed in any order that is physically possible.

[0215] All cited patents, patent publications, and non-patent references are incorporated herein by reference in their entirety.

[0216] Although the embodiments have been disclosed, the present invention is not limited thereto.

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Claims

1. A method for treating cancer in a patient, comprising the steps of: The collagen device having the DDR1 peptide is introduced into the patient's body, thereby delivering the DDR1 peptide to the cancer.

2. The method of claim 1, wherein the collagen device is an injectable device comprising one or more of type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites, wherein the type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites comprise a DDR1 peptide.

3. The method of claim 1, wherein the collagen device is an inlay device comprising one or more of type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites, wherein the type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites comprise a DDR1 peptide.

4. The method of claim 1, wherein the collagen device is a membrane comprising one or more of type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites, wherein the type III collagen, native rh type III collagen, and rh type III collagen modified with DDR1 binding sites comprise a DDR1 peptide. The method of claim 1 , wherein the DDR1 peptide comprises the DDR1 peptide dispersed in a degradable carrier.

6. The method according to claim 5, wherein the carrier is selected from gelatin and other collagens.

7. The method of claim 5, wherein the carrier is a synthetic degradable polymer.

8. The method of claim 5, wherein the carrier is selected from the group consisting of PEG, PLA, PGA, PLGA, and polymers thereof.

9. The method of claim 1, wherein the DDR1 peptide is dispersed in an injectable carrier.

10. The method according to claim 1, wherein the DDR1 peptide is selected from a. Peptide containing the GVMGFO sequence; b. Peptide containing GVMGFP sequence; c. Peptides containing the GPSGFO sequence; d. Peptide containing GPSGFP sequence; e. Peptides containing the GPRGFO sequence; f. A peptide containing a GPRGFP sequence; g. Peptides containing the GARGFO sequence; h. Peptide containing GARGFP sequence; i. Peptides containing the GQOGFO sequence; j. Peptide containing the GQOGFP sequence; k. Peptides containing the GAMGFO sequence; 1. Peptide containing GAMGFP sequence; m. A peptide comprising the sequence of one of peptides al and flanked by at least one amino acid triplet; n. a peptide sequence that performs a function selected from binding to DDR1, inducing autophosphorylation of cell membrane-bound DDR1, and activating a DDR1 signaling pathway; o. A peptide according to one of the peptides an, wherein the form is selected from a single helical form, a double helical form and a triple helical form; p. a peptide according to one of the peptides ai, which has been modified with positively charged amino acid side chains; and q. A peptide according to peptide ai which has been modified with negatively charged amino acid side chains.

11. The method of claim 10, wherein the at least one amino acid triplet is selected from the group consisting of a GPO amino acid triplet and a GfO amino acid triplet, wherein O is the amino acid hydroxyproline and f is 2S,4S fluoroproline.

12. The method of claim 11, wherein the at least one amino acid triplet is selected from GPO(n)-GVMGFO-GPO(n) and GfO(n)-GVMGFO-GfO(n), wherein n is the number of GPO or GfO repeats.

13. A method for treating cancer in a patient, comprising the steps of: The DDR1 peptide is introduced into the patient, thereby delivering the DDR1 peptide to the cancer.

14. The method of claim 13, wherein the DDR1 peptide comprises the DDR1 peptide dispersed in a degradable carrier.

15. The method of claim 14, wherein the carrier is selected from gelatin and other collagens.

16. The method of claim 14, wherein the carrier is a synthetic degradable polymer.

17. The method of claim 14, wherein the carrier is selected from the group consisting of PEG, PLA, PGA, PLGA, and polymers thereof.

18. The method of claim 13, wherein the DDR1 peptide is dispersed in an injectable carrier.

19. The method of claim 13, wherein the DDR1 peptide is selected from a. A peptide containing the sequence GVMGFO; b. Peptide containing GVMGFP sequence; c. Peptides containing the GPSGFO sequence; d. Peptide containing GPSGFP sequence; e. Peptides containing the GPRGFO sequence; f. A peptide sequence containing GPRGFP; g. Peptides containing the GARGFO sequence; h. Peptide containing GARGFP sequence; i. Peptides containing the GQOGFO sequence; j. Peptide containing the GQOGFP sequence; k. Peptides containing the GAMGFO sequence; 1. Peptide containing GAMGFP sequence; m. A peptide comprising the sequence of one of peptides al and flanked by at least one amino acid triplet; n. a peptide sequence that performs a function selected from binding to DDR1, inducing autophosphorylation of cell membrane-bound DDR1, and activating a DDR1 signaling pathway; o. A peptide according to one of the peptides an, wherein the form is selected from a single helical form, a double helical form and a triple helical form; p. a peptide according to one of the peptides ai, which has been modified with positively charged amino acid side chains; and q. A peptide according to peptide ai which has been modified with negatively charged amino acid side chains.

20. The method of claim 19, wherein the at least one amino acid triplet is selected from the group consisting of a GPO amino acid triplet and a GfO amino acid triplet, wherein O is the amino acid hydroxyproline and f is 2S,4S fluoroproline.

21. The method of claim 20, wherein the at least one amino acid triplet is selected from GPO(n)-GVMGFO-GPO(n) and GfO(n)-GVMGFO-GfO(n), wherein n is the number of GPO or GfO repeats.

22. A method for treating cancer in a patient, comprising the steps of: introducing a DDR1 peptide into a patient, thereby delivering the DDR1 peptide to the cancer; and administering an anti-cancer therapeutic agent to the patient.

23. The method of claim 22, wherein the anti-cancer therapeutic is selected from the group consisting of a cancer API; a cancer vaccine; and an inhibitor / degrader of HPK1.

24. A method for treating cancer in a patient, comprising the steps of: introducing a collagen device having a DDR1 peptide into a patient, thereby delivering the DDR1 peptide to the cancer; as well as An anti-cancer therapeutic is administered to the patient.

25. The method of claim 24, wherein the anti-cancer therapeutic is selected from the group consisting of a cancer API; a cancer vaccine; and an inhibitor / degrader of HPK1.

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