Therapeutic constructs for co-delivery of mitotic kinase inhibitor and immune checkpoint inhibitor

Abstract

Disclosed herein are therapeutic constructs including a delivery particle, at least one mitotic kinase inhibitor, and at least one immune checkpoint inhibitor. Also disclosed are therapeutic constructs including a mitotic kinase inhibitor, an immune checkpoint inhibitor, and a chemical linker. These therapeutic constructs cause cancer death by both therapeutic and immune effects and promote targeted delivery of more therapeutics to the surviving cancer cells in a positive feed-back loop. They enhance therapeutic index of free drugs and can be used intratumorally or systemically. This strategy can treat broad cancer types and is particular useful for cancer without obvious receptors for cancer-targeted delivery of otherwise toxic therapeutics.

What is claimed is: 
     
         1 . A therapeutic construct comprising:
 a delivery system comprising   at least one mitotic kinase inhibitor; and   at least one immune checkpoint inhibitor.   
     
     
         2 . The therapeutic construct of  claim 1 , wherein the delivery system comprises a liposome, a lipid-based particle, a polymeric particle, an inorganic or organic nanoparticle or microparticle, or a hybrid thereof. 
     
     
         3 . The therapeutic construct of  claim 2 , wherein the delivery vehicle comprises one or more of fullerenes, endohedral metallofullerenes, trimetallic nitride templated endohedral metallofullerenes, single-walled and multi-walled carbon nanotubes, calcium phosphate particles, aluminum salt particles, branched and dendritic carbon nanotubes, gold nanorods, silver nanorods, single-walled and multi-walled boron/nitrate nanotubes, carbon nanotube peapods, carbon nanohorns, carbon nanohorn peapods, liposomes, nanoshells, dendrimers, microparticles, quantum dots, superparamagnetic nanoparticles, nanorods, cellulose nanoparticles, silicon, silica and polymer micro- and nano-spheres, silica-shells, biodegradable PLGA micro- and nano-spheres, gold nanoparticles, cerium oxide particles, zinc oxide particles, silver nanoparticles, carbon nanoparticles, iron nanoparticles, and/or modified micelles. 
     
     
         4 . The therapeutic construct of any one of  claims 1 - 3 , wherein the delivery vehicle comprises a mesoporous silica nanoparticle. 
     
     
         5 . The therapeutic construct of  claim 4 , wherein the mesoporous silica nanoparticle has a mean particle size of about 5-200 nm. 
     
     
         6 . The therapeutic construct of  claim 4  or  5 , wherein the mesoporous silica nanoparticle is coated with cross-linked polyethyleneimine and polyethylene glycol. 
     
     
         7 . The therapeutic construct of any one of  claims 1 - 6 , wherein the at least one mitotic kinase inhibitor and/or immune checkpoint inhibitor comprises an oligonucleotide, a polynucleotide, a small molecule inhibitor, or an antibody. 
     
     
         8 . The therapeutic construct of any one of  claims 1 - 7 , wherein the at least one mitotic kinase inhibitor is an inhibitor of a polo-like kinase (PLK), an Aurora kinase, cyclin-dependent kinase (CDK)1, CDK2, HASPIN, monopolar spindle 1 kinase (Mps1), a NimA-related kinase (NEK). 
     
     
         9 . The therapeutic construct of any one of  claims 1 - 8 , wherein the at least one mitotic kinase inhibitor comprises one or more of GSK461364, BI2536, Tak960, NMS-P937, volasertib, Chk 1 Kinase Inhibitor LY2603618, AU14022, YK-4-279, AZ703, alisertib, prexasertib, or AZD7762. 
     
     
         10 . The therapeutic construct of any one of  claims 1 - 9 , wherein the at least one mitotic kinase inhibitor comprises volasertib. 
     
     
         11 . The therapeutic construct of any one of  claims 1 - 10 , wherein the at least one immune checkpoint inhibitor comprises a siRNA, inhibitor, or antibody against one or more of PD-L1, PD-1, TIM-3, LAG-3, or CTLA-4. 
     
     
         12 . The therapeutic construct of any one of  claims 1 - 11 , wherein the at least one immune checkpoint inhibitor is an antibody against PD-L1, PD-1, or CTLA-4. 
     
     
         13 . The therapeutic construct of any one of  claims 1 - 12 , wherein the at least one immune checkpoint inhibitor is an antibody against PD-1. 
     
     
         14 . The therapeutic construct of  claim 13 , wherein the at least one immune checkpoint inhibitor comprises at least one of: nivolumab, pembrolizumab, MPDL3280A, ipilimumab, tremelimumab, atezolizumab, avelumab, durvalumab, cemiplimab, pidilizumab, or spartalizumab. 
     
     
         15 . The therapeutic construct of any of the previous claims, further comprising an adjuvant. 
     
     
         16 . The therapeutic construct of  claim 15 , wherein the adjuvant comprises one or more of a CpG oligonucleotide, a DNA TLR agonist containing a CpG sequence, a non-CpG DNA TLR agonist, an RNA TLR agonist, an aluminum salt, an anti-CD40 antibody, a fusion protein, a cytokine, a small molecule TLR agonist, an oil- or surfactant-based adjuvant, a lipopolysaccharide, a plant extract, or a derivative thereof. 
     
     
         17 . The therapeutic construct of  claim 15  or  16 , wherein the adjuvant comprises a CpG oligonucleotide, imiquimod, resiquimod, gardiquimod, poly I:C, poly ICLC, dSLIM, or EnanDIM. 
     
     
         18 . The therapeutic construct of  claim 16 , wherein the adjuvant comprises a CpG oligonucleotide. 
     
     
         19 . The therapeutic construct of any one of  claims 1 - 18 , having a hydrodynamic size of 5-999 nm. 
     
     
         20 . The therapeutic construct of any one of  claims 1 - 18 , having a hydrodynamic size of 1-1000 microns. 
     
     
         21 . A therapeutic construct comprising:
 an immune checkpoint inhibitor;   a mitotic kinase inhibitor; and   a chemical linker linking the immune checkpoint inhibitor and the mitotic kinase inhibitor.   
     
     
         22 . The therapeutic construct of  claim 21 , wherein the mitotic kinase inhibitor is an oligonucleotide, a polynucleotide, a small molecule inhibitor, or an antibody. 
     
     
         23 . The therapeutic construct of  claim 21  or  22 , wherein the immune checkpoint inhibitor is an oligonucleotide, a polynucleotide, a small molecule inhibitor, or an antibody. 
     
     
         24 . The therapeutic construct of any one of  claims 21 - 23 , wherein the immune checkpoint inhibitor is an antibody. 
     
     
         25 . The therapeutic construct of any one of  claims 21 - 24 , wherein the immune checkpoint inhibitor is an antibody against PD-L1, PD-1, TIM-3, LAG-3, or CTLA-4. 
     
     
         26 . The therapeutic construct of any one of  claims 21 - 25 , wherein the immune checkpoint inhibitor is an antibody against PD-L1, PD-1, or CTLA-4. 
     
     
         27 . The therapeutic construct of any one of  claims 21 - 26 , wherein the immune checkpoint inhibitor is an antibody against PD-L1. 
     
     
         28 . The therapeutic construct of any one of  claims 21 - 27 , wherein the mitotic kinase inhibitor is selected from GSK461364, BI2536, Tak960, NMS-P937, volasertib, Chk 1 Kinase Inhibitor LY2603618, AU14022, YK-4-279, AZ703, alisertib, prexasertib, or AZD7762. 
     
     
         29 . The therapeutic construct of any one of  claims 21 - 28 , wherein the mitotic kinase inhibitor is alisertib. 
     
     
         30 . The therapeutic construct of any one of  claims 21 - 29 , wherein the chemical linker comprises one or more of the a hydrazine; a disulfide; N-succinimidyl-4-(2-pyridyldithio)butanoate; N-succinimidyl-4-(2-pyridyldithio)-2-sulfo butanoate; perfluorophenyl 3-(pyridin-2-yldisulfanyl)propanoate; 2,5-dioxopyrrolidin-1-yl 3-methyl-3-(pyridin-2-yldisulfanyl)butanoate; Gly-Phe-Leu-Gly; Ala-Leu-Ala-Leu; Val-Cit; Phe-Lys; Val-Ala; Ala-Phe-Lys; Phe-Lys; (Gly) n , wherein n is 1-20; a β-glucuronide linker; maleimidocaproyl; N-(maleimidomethyl)cyclohexane-1-carboxylate; 4-(4-acetylphenoxy)butanoic acid; dibromomaleimide; para-aminobenzoic acid; 4-nitrophenol; acetic acid; formic acid; 4-maleimidobutyric acid N-succinimidyl ester; N-(4-maleimidobutyryloxy)succinimide; N-(6-maleimidocaproyloxy)succinimide; 3-maleimidopropionic acid N-succinimidyl ester; N-(3-maleimidopropionyloxy)succinimide; 5-maleimidovalericacid-NHS; linear, branched, or multi-arm polyethylene glycol having a molecular weight of 100-10000 Da; propargyl-N-hydroxysuccinimidyl ester; pyrophosphate; succimimidyl-4-azidobutyrate; 4-azidobenzoic acid N-hydroxysuccinimide ester; tert-butyl 1-(4-formylphenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-oate; or a residue thereof. 
     
     
         31 . The therapeutic construct of any one of  claims 21 - 30 , wherein the chemical linker comprises N-(maleimidomethyl)cyclohexane-1-carboxylate linker or a residue thereof. 
     
     
         32 . The therapeutic construct of any one of  claims 21 - 31 , wherein the chemical linker comprises sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate. 
     
     
         33 . The therapeutic construct of any one of  claims 21 - 32 , wherein the chemical linker comprises a linear polyethyleneglycol having a molecular weight of 100-10000 Da, or a residue thereof. 
     
     
         34 . The therapeutic construct of any one of  claims 21 - 33 , wherein the ratio of mitotic kinase inhibitor to immune checkpoint inhibitor is about 1-20. 
     
     
         35 . The therapeutic construct of  claim 34 , wherein the ratio of mitotic kinase inhibitor to immune checkpoint inhibitor is about 2-8. 
     
     
         36 . The therapeutic construct of  claim 34  or  35 , wherein the ratio of mitotic kinase inhibitor to immune checkpoint inhibitor is about 4-6. 
     
     
         37 . The therapeutic construct of any one of  claims 21 - 33 , wherein the ratio of immune checkpoint inhibitor to mitotic kinase inhibitor is about 1-20. 
     
     
         38 . The therapeutic construct of  claim 37 , wherein the ratio of immune checkpoint inhibitor to mitotic kinase inhibitor is about 2-8. 
     
     
         39 . The therapeutic construct of  claim 37  or  38 , wherein the ratio of immune checkpoint inhibitor to mitotic kinase inhibitor is about 4-6. 
     
     
         40 . A composition comprising the therapeutic construct of any one of  claims 1 - 39  and a pharmaceutically acceptable carrier, excipient, or diluent. 
     
     
         41 . A method of treating cancer comprising administering to a subject with cancer an effective amount of the therapeutic construct of any one of  claims 1 - 39 , or the composition of  claim 40 . 
     
     
         42 . The method of  claim 41 , wherein the subject is a human. 
     
     
         43 . A method of treating a cell exhibiting symptoms of cancer comprising contacting the cell with a therapeutically effective amount of the therapeutic construct of any one of  claims 1 - 39 , or a composition of  claim 40 . 
     
     
         44 . A method of treating a cell obtained from a subject exhibiting symptoms of cancer, comprising contacting the cell with a therapeutically effective amount of the therapeutic construct of any one of  claims 1 - 39 , or the composition of  claim 40 . 
     
     
         45 . A method of treating a cell obtained from a subject exhibiting symptoms of cancer, comprising contacting cell ex vivo with a therapeutically effective amount of the therapeutic construct of any one of  claims 1 - 39 , or the composition of  claim 40 . 
     
     
         46 . The method of  claim 44  or  45 , wherein the cell is a cancer cell. 
     
     
         47 . The method of  claim 44  or  45 , wherein the cell is not a cancer cell. 
     
     
         48 . The method of  claim 47 , wherein the cell is an immune cell. 
     
     
         49 . The method of any one of  claims 41 - 48 , further comprising administering at least one treated cell back to a subject. 
     
     
         50 . A method of treating a subject diagnosed as having a hyperproliferative disease or condition, comprising administering to the subject an effective amount of the composition of  claim 40 . 
     
     
         51 . The method of  claim 50 , wherein the hyperproliferative disease comprises one or more of cancer, precancer, or cancer metastasis. 
     
     
         52 . The method of  claim 50  or  51 , wherein the hyperproliferative disease comprises one or more of melanoma, lung cancer, breast cancer, pancreatic cancer, brain cancer, prostate cancer, head and neck cancer, kidney cancer, colorectal cancer, lymphoma, colon cancer, or liver cancer. 
     
     
         53 . The method of any one of  claims 50 - 52 , wherein the administering comprises one or more of:
 injection to or at a tumor in the subject;   infusion locally to or at a tumor in the subject;   systemic injection in the subject;   systemic infusion in the subject;   inhalation by the subject;   oral administration to the subject; or   topical application to the subject.   
     
     
         54 . The method of any one of  claims 50 - 53 , wherein the administering comprises microneedle application. 
     
     
         55 . A method of enhancing an effect of an anti-cancer therapy in a subject in need thereof, comprising administering to a subject in need thereof:
 an effective amount of the therapeutic construct of any one of  claims 1 - 39 , or the composition of  claim 40 ; and   at least one anti-cancer agent.   
     
     
         56 . The method of  claim 55 , wherein the anti-cancer agent is a chemotherapeutic agent, a targeted therapeutic agent, or an immune checkpoint inhibitor. 
     
     
         57 . The method of  claim 55  or  56 , wherein the therapeutic construct or composition and the anti-cancer therapy are administered sequentially or concurrently. 
     
     
         58 . A method of enhancing, increasing, or improving a radiation therapy effect in a subject diagnosed as having a neoplasia, comprising administering to a subject in need thereof:
 an effective amount of the therapeutic construct of any one of  claims 1 - 39 , or the composition of  claim 40 ; and   at least one radiation therapy.   
     
     
         59 . The method of  claim 58 , wherein the therapeutic construct or composition and the radiation therapy are administered sequentially or concurrently. 
     
     
         60 . The method of any one of  claims 49 - 59 , wherein the subject is human. 
     
     
         61 . A kit comprising:
 the therapeutic construct of any one of  claims 1 - 39 ; and   at least one anti-cancer agent.   
     
     
         62 . The kit of  claim 61 , wherein the anti-cancer agent is a chemotherapeutic agent, a targeted therapeutic agent, or an immune checkpoint inhibitor.