Truncated car peptides and methods and compositions using truncated car peptides

Abstract

Disclosed are compositions and methods useful for targeting and internalizing molecules into cells of interest and for penetration by molecules of tissues of interest. The compositions and methods are based on peptide sequences, such as truncated CAR peptides, that are selectively internalized by a cell, penetrate tissue, or both. The disclosed internalization and tissue penetration is useful for delivering therapeutic and detectable agents to cells and tissues of interest.

1 . An isolated peptide, wherein the C-terminal end of the peptide consists of the amino acid sequence CARSKNK (SEQ ID NO:4). 
     
     
         2 . The peptide of  claim 1 , wherein the peptide is a modified peptide. 
     
     
         3 - 5 . (canceled) 
     
     
         6 . The peptide of  claim 1 , wherein the peptide is an activatable peptide. 
     
     
         7 . The peptide of  claim 6 , wherein the amino acid sequence CARSKNK (SEQ ID NO:4) at the C-terminal end of the peptide is blocked by a blocking group coupled to the terminal carboxy group. 
     
     
         8 . The peptide of  claim 7 , wherein the blocking group comprises an amino acid or an amino acid sequence. 
     
     
         9 . A composition comprising the peptide of  claim 1 . 
     
     
         10 . The composition of  claim 9  further comprising a co-composition, wherein the peptide and the co-composition are not covalently coupled or non-covalently associated with each other. 
     
     
         11 . The composition of  claim 9  further comprising a cargo composition, wherein the peptide and the cargo composition are covalently coupled or non-covalently associated with each other. 
     
     
         12 . The composition of  claim 9 , wherein the peptide selectively homes to regenerating tissue, a site of injury, a surgical site, a tumor, tumor vasculature, a site of tumor angiogenesis, a site of inflammation, a site of arthritis, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, or interstitial space of lungs. 
     
     
         13 . (canceled) 
     
     
         14 . The composition of  claim 10 , wherein the co-composition selectively homes to regenerating tissue, a site of injury, a surgical site, a tumor, tumor vasculature, a site of tumor angiogenesis, a site of inflammation, a site of arthritis, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, or interstitial space of lungs. 
     
     
         15 . The composition of  claim 10 , wherein the peptide and the co-composition are not bound to each other. 
     
     
         16 - 18 . (canceled) 
     
     
         19 . The composition of  claim 10 , wherein the co-composition comprises a therapeutic agent, a therapeutic protein, a therapeutic compound, a therapeutic composition, a chemotherapeutic agent, a cancer chemotherapeutic agent, a toxin, a cytotoxic agent, Abraxane, paclitaxel, taxol, imatinib, an anti-angiogenic agent, a pro-angiogenic agent, an anti-inflammatory agent, an anti-arthritic agent, a TGF-β inhibitor, decorin, a systemic vasodilator, an anti-coagulant, tissue factor pathway inhibitor (TFPI), site-inactivated factor VIIa, a β-2 agonist, salmeterol, formoterol, N-Acetylcysteine (NAC), Superoxide Dismutase (SOD), a superoxide dismutase mimetic, EUK-8, an endothelin (ET-1) receptor antagonist, a prostacyclin derivative, a phosphodiesterase type 5 inhibitor, Ketoconazole, a small interfering RNA (siRNA), a microRNA (miRNA), a polypeptide, a nucleic acid molecule, a small molecule, a carrier, a vehicle, a virus, a phage, a viral particle, a phage particle, a viral capsid, a phage capsid, a virus-like particle, a liposome, a micelle, a bead, a nanoparticle, a microparticle, a detectable agent, a contrast agent, an imaging agent, a label, a labeling agent, a fluorophore, fluorescein, rhodamine, FAM, a radionuclide, indium-111, technetium-99, carbon-11, carbon-13, or a combination. 
     
     
         20 . The composition of  claim 11 , wherein the peptide is comprised in a tCAR composition. 
     
     
         21 . The composition of  claim 20 , wherein the tCAR composition comprises one or more cargo compositions. 
     
     
         22 . The composition of  claim 20 , wherein the tCAR composition comprises one or more homing molecules. 
     
     
         23 . The composition of  claim 11 , wherein the peptide is comprised in a tCAR conjugate. 
     
     
         24 . The composition of  claim 23 , wherein the tCAR conjugate comprises one or more cargo compositions. 
     
     
         25 . The composition of  claim 10 , wherein the tCAR conjugate comprises one or more homing molecules. 
     
     
         26 . The composition of  claim 10 , wherein the composition comprises a plurality of cargo compositions. 
     
     
         27 . The composition of  claim 10 , wherein the composition comprises a plurality of copies of the peptide. 
     
     
         28 . The composition of  claim 10 , wherein the composition comprises a plurality of co-compositions. 
     
     
         29 . The composition of  claim 9 , wherein the composition further comprises a surface molecule and a plurality of membrane perturbing molecules. 
     
     
         30 . The composition of  claim 29 , wherein the composition further comprises one or more homing molecules, wherein the homing molecules selectively home to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis. 
     
     
         31 . The composition of  claim 10 , wherein the co-composition comprises a surface molecule, one or more homing molecules, and a plurality of membrane perturbing molecules, wherein the homing molecules selectively home to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis. 
     
     
         32 . The composition of  claim 11 , wherein the cargo composition comprises a surface molecule, one or more homing molecules, and a plurality of membrane perturbing molecules, wherein the homing molecules selectively home to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis. 
     
     
         33 - 39 . (canceled) 
     
     
         40 . The composition of  claim 9 , wherein the composition is internalized in cells. 
     
     
         41 . The composition of  claim 9 , wherein the composition penetrates tissue. 
     
     
         42 - 55 . (canceled) 
     
     
         56 . The composition of  claim 9  further comprising one or more moieties. 
     
     
         57 . The composition of  claim 56 , wherein the moieties are independently selected from the group consisting of a therapeutic agent, a therapeutic protein, a therapeutic compound, a therapeutic composition, a chemotherapeutic agent, a cancer chemotherapeutic agent, a toxin, a cytotoxic agent, Abraxane, paclitaxel, taxol, imatinib, an anti-angiogenic agent, a pro-angiogenic agent, an anti-inflammatory agent, an anti-arthritic agent, a TGF-β inhibitor, decorin, a systemic vasodilator, an anti-coagulant, tissue factor pathway inhibitor (TFPI), site-inactivated factor VIIa, a β-2 agonist, salmeterol, formoterol, N-Acetylcysteine (NAC), Superoxide Dismutase (SOD), a superoxide dismutase mimetic, EUK-8, an endothelin (ET-1) receptor antagonist, a prostacyclin derivative, a phosphodiesterase type 5 inhibitor, Ketoconazole, a small interfering RNA (siRNA), a microRNA (miRNA), a polypeptide, a nucleic acid molecule, a small molecule, a carrier, a vehicle, a virus, a phage, a viral particle, a phage particle, a viral capsid, a phage capsid, a virus-like particle, a liposome, a micelle, a bead, a nanoparticle, a microparticle, a detectable agent, a contrast agent, an imaging agent, a label, a labeling agent, a fluorophore, fluorescein, rhodamine, FAM, a radionuclide, indium-111, technetium-99, carbon-11, carbon-13, or a combination. 
     
     
         58 . The composition of  claim 9 , wherein the tCAR composition has a therapeutic effect. 
     
     
         59 . The composition of  claim 10 , wherein the co-composition has a therapeutic effect. 
     
     
         60 . The composition of  claim 58 , wherein the therapeutic effect is a slowing in the increase of or a reduction of tumor burden. 
     
     
         61 . The composition of  claim 58 , wherein the therapeutic effect is a slowing of the increase of or reduction of tumor size. 
     
     
         62 . The composition of  claim 58 , wherein the therapeutic effect comprises a reduction in inflammation, an increase in speed of wound healing, reduction in amounts of scar tissue, decrease in pain, decrease in swelling, or decrease in necrosis. 
     
     
         63 . The composition of  claim 58 , wherein the therapeutic effect comprises pulmonary vasodilation, decrease in pulmonary pressure, anti-coagulation, airway smooth muscle relaxation, increase in glutathione (GSH), decrease in inflammatory immune response, inhibition of thromboxane synthesis, or inhibition of leukotriene synthesis. 
     
     
         64 . A method of enhancing internalization, penetration, or both into or through a cell, tissue, or both, the method comprising:
 exposing the cell, tissue, or both to a tCAR composition, thereby enhancing internalization, penetration, or both into or through the cell, tissue, or both,   wherein the tCAR composition comprises the peptide of  claim 1 .   
     
     
         65 . A method of enhancing internalization, penetration, or both into or through a cell, tissue, or both, the method comprising:
 exposing the cell, tissue, or both to a tCAR composition, thereby enhancing internalization, penetration, or both into or through the cell, tissue, or both,   wherein the tCAR composition comprises the composition of  claim 9 .   
     
     
         66 . The method of  claim 64  further comprising exposing the cell, tissue, or both to a co-composition, thereby enhancing internalization, penetration, or both of the co-composition into or through the cell, tissue, or both, wherein, prior to exposing the cell, tissue, or both, the tCAR composition and the co-composition are not covalently coupled or non-covalently associated with each other. 
     
     
         67 . The method of  claim 64  further comprising exposing the cell, tissue, or both to a cargo composition, thereby enhancing internalization, penetration, or both of the cargo composition into or through the cell, tissue, or both, wherein the tCAR composition and the cargo composition are covalently coupled or non-covalently associated with each other. 
     
     
         68 . The method of  claim 65 , wherein the cell, tissue, or both is in a subject. 
     
     
         69 . The method of  claim 68 , wherein the cell, tissue, or both are exposed to the tCAR composition by administering the tCAR composition to the subject. 
     
     
         70 . The method of  claim 66 , wherein the cell, tissue, or both is in a subject, wherein the cell, tissue, or both are exposed to the tCAR composition and the co-composition by administering the tCAR composition and the co-composition to the subject. 
     
     
         71 . The method of  claim 67 , wherein the cell, tissue, or both is in a subject, wherein the cell, tissue, or both are exposed to the tCAR composition and the cargo composition by administering the tCAR composition and the cargo composition to the subject. 
     
     
         72 - 75 . (canceled) 
     
     
         76 . The method of  claim 70 , wherein the tCAR composition and the co-composition are administered to the subject simultaneously. 
     
     
         77 . The method of  claim 76 , wherein the tCAR composition and the co-composition are administered to the subject in a single composition comprising the tCAR composition and the co-composition. 
     
     
         78 . The method of  claim 70 , wherein the tCAR composition and the co-composition are administered to the subject in separate compositions. 
     
     
         79 . The method of  claim 70 , wherein the tCAR composition and the co-composition are administered to the subject at different times. 
     
     
         80 . The method of  claim 79 , wherein the tCAR composition and the co-composition are administered to the subject in separate compositions. 
     
     
         81 . The method of  claim 78 , wherein the tCAR composition and the co-composition are administered to the subject by separate routes. 
     
     
         82 . The method of  claim 66 , wherein the tCAR composition and the co-composition are not bound to each other. 
     
     
         83 - 94 . (canceled) 
     
     
         95 . The method of  claim 68 , wherein the subject has a disease or condition. 
     
     
         96 . The method of  claim 95 , wherein the disease is pulmonary or fibrotic. 
     
     
         97 . The method of  claim 96 , wherein the disease or condition is pulmonary arterial hypertension (PAH). 
     
     
         98 . The method of  claim 95 , wherein the disease or condition is cancer. 
     
     
         99 . The method of  claim 95 , wherein the disease or condition is an autoimmune disease. 
     
     
         100 . The method of  claim 95 , wherein the disease or condition is an inflammatory disease. 
     
     
         101 . The method of  claim 68 , wherein the subject has one or more sites to be targeted, wherein the tCAR composition homes to one or more of the sites to be targeted. 
     
     
         102 . The method of  claim 68 , wherein the subject has one or more sites to be targeted, wherein the co-composition or cargo composition homes to one or more of the sites to be targeted. 
     
     
         103 . The method of  claim 64 , wherein the peptide selectively homes to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis. 
     
     
         104 . The method of  claim 64 , wherein the tCAR composition selectively homes to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis. 
     
     
         105 . The method of  claim 66 , wherein the co-composition or cargo composition selectively homes to tumor vasculature, lung vasculature, regenerating tissue, wounded tissue, angiogenic tissue, cancer issue, lung tissue, pulmonary arterial hypertension lung vasculature, pulmonary arterial hypertension lesions, remodeled pulmonary arteries, interstitial space of lungs, a site of injury, a surgical site, a tumor, a site of angiogenesis, a site of tumor angiogenesis, a site of inflammation, or a site of arthritis.