JP2014505666A - Antihypertensive composition and use for cancer treatment - Google Patents

Abstract

Disclosed are methods and compositions for improving the delivery and / or efficacy of cancer therapeutics. By administering an antihypertensive agent to a subject as a single agent or in combination with a cancer therapeutic agent (e.g., a therapeutic agent ranging in size from a large nano-therapeutic agent to a low molecular weight chemotherapeutic agent and / or an oxygen radical) Disclosed are methods and compositions for treating or preventing cancer (eg, solid tumors such as fibrogenic tumors).

Description

Cross-reference to related applications This application is based on 35 U.SC §119 based on US Provisional Application No. 61 / 415,192 filed on November 18, 2010; and US Provisional Application No. 61 / 438,240 filed on January 31, 2011. Claim the benefits of (e). Each of which is hereby incorporated by reference in its entirety.

Government Assistance This invention was made with federal funding under grant number PO1-CA-80124-03 awarded by the National Institutes of Health. The US government has certain rights in the invention.

Background Advances in biomedical research have led to the introduction of several new systemically administered molecular and nanotherapeutics in both preclinical and clinical settings (Jones, D. (2007) Nat Rev Drug Discov 6,174-175 (Non-Patent Document 1); Moghimi, SM et al. (2005) Faseb J 19,311-330 (Non-Patent Document 2)). Although these new agents act on unique targets that provide greater specificity or improved pharmacodynamic properties for tumor cells, their effectiveness is limited by delivery due to the characteristics of the tumor microenvironment (Jain, RK (1998) Nat Med 4,655-657 (Non-patent Document 3); Sanhai, WR et al. (2008) Nat Nanotechnol 3,242-244 (Non-patent Document 4)). For example, FDA-approved PEGylated liposomal doxorubicin (DOXIL®) and oncolytic viruses for which multiple clinical trials are currently underway have intratumoral dispersion and therapeutic efficacy in size (approximately 100 nm) (Nemunaitis J, et al. (2001) J Clin Oncol 19: 289-298 (Non-patent Document 5)).

  In tumors, at least two processes governing drug delivery after systemic administration, namely, vascular transport in tissues and transvascular transport into tissues, are hindered by physiological barriers for all therapeutic drug classes (Jain , RK & Stylianopoulos (2010) Nat Rev Clin Oncol. 139 (Non-patent document 6); Chauhan, VP et al. (2009) Biophysical journal 97, 330-336 (Non-patent document 7)). These barriers include pancreatic cancer (Olive, KP et al. (2009) Science 324,1457-1461), colorectal cancer (Halvorsen, TB & Seim, E. (1989) J Clin Fibrogenic tumors such as Pathol 42,162-166 (Non-patent document 9), lung cancer and breast cancer (Ronnov-Jessen, L. et al. (1996) Physiol Rev 76,69-125 (Non-patent document 10)), Affects treatment for patients with fibrous tumors. Fibrous tumors typically have a dense collagen network, so small fiber spacing in the stroma slows the movement of particles larger than 10 nanometers (Netti PA, et al. (2000) Cancer Res 60: 2497-2503 (Non-Patent Document 11); Pluen A, et al. (2001) Proc Natl Acad Sci USA 98: 4628-4633 (Non-Patent Document 12); Ramanujan S, et al. (2002) Biophys J 83 : 1650-1660 (Non-Patent Document 13); and Brown E, et al. (2003) Nat Med 9: 796-800 (Non-Patent Document 14)). These barriers limit the amount of drug reaching the target cancer cells, resulting in low drug efficacy.

  Currently, approaches to overcome these delivery barriers for nanotherapeutics and low molecular weight drugs are limited. Thus, new cancer treatments, especially new ones that enhance the delivery and dispersion of cancer therapies, including nanotherapeutic agents (eg, lipid or polymer nanoparticles and viruses), protein drugs, nucleic acid drugs, and small molecule chemotherapeutic agents There is a need to identify new agents.

Jones, D. (2007) Nat Rev Drug Discov 6,174-175 Moghimi, S.M.et al. (2005) Faseb J 19,311-330 Jain, R.K. (1998) Nat Med 4,655-657 Sanhai, W.R.et al. (2008) Nat Nanotechnol 3,242-244 Nemunaitis J, et al. (2001) J Clin Oncol 19: 289-298 Jain, R.K. & Stylianopoulos (2010) Nat Rev Clin Oncol. 139 Chauhan, V.P. et al. (2009) Biophysical journal 97,330-336 Olive, K.P.et al. (2009) Science 324,1457-1461 Halvorsen, T.B. & Seim, E. (1989) J Clin Pathol 42,162-166 Ronnov-Jessen, L. et al. (1996) Physiol Rev 76, 69-125 Netti PA, et al. (2000) Cancer Res 60: 2497-2503 Pluen A, et al. (2001) Proc Natl Acad Sci USA 98: 4628-4633 Ramanujan S, et al. (2002) Biophys J 83: 1650-1660 Brown E, et al. (2003) Nat Med 9: 796-800

  The present invention is based, in part, on the discovery that losartan, an angiotensin II receptor antagonist drug approved for the treatment of hypertension (hypertension), improves the delivery and efficacy of cancer therapeutics. We, among others, losartan normalizes collagen, the stromal matrix of solid tumors, and facilitates the dispersion and / or penetration of chemotherapeutic drugs, including high molecular weight chemotherapeutic drugs, such as nanotherapeutic drugs I discovered that. For example, losartan reduces collagen I levels in cancer-associated fibroblasts (CAF) isolated from breast cancer biopsies (eg, reduces collagen production) and reduces human breast, pancreas, and skin in mice It caused a dose-dependent decrease in matrix collagen in a tumor fibrosis model. Losartan also improved the dispersion, therapeutic efficacy, and / or penetration of nanoparticles (eg, oncolytic herpes simplex virus (HSV) and PEGylated liposomal doxorubicin (DOXIL®)). We have also found that losartan facilitates vascular decompression and vascular normalization, improves tumor perfusion and delivery of low molecular weight chemotherapeutic drugs, and thus enhances the therapeutic effects of radiation and chemotherapeutic drugs. discovered.

  Accordingly, methods and compositions for improving the delivery and / or efficacy of therapeutic agents (eg, cancer therapeutic agents) are disclosed. Targeting antihypertensive and / or collagen modifying agents as a single agent or in combination with therapeutic agents (eg, cancer therapeutic agents of sizes ranging from large nanotherapeutics to low molecular weight chemotherapeutics and / or oxygen radicals) Disclosed are methods and compositions for treating or preventing cancer (e.g., solid tumors such as fibrogenic tumors) by administering to.

  Accordingly, in one aspect, the present invention improves the method of treating or preventing a hyperproliferative disorder (eg, cancer) in a subject or the delivery and / or efficacy of a therapy (eg, cancer therapy) to a subject. Features how to do. The method comprises a condition sufficient to treat or prevent a disorder (eg, a cancer or tumor) in a subject, or to improve delivery and / or efficacy of a therapy (eg, cancer therapy) provided to the subject, For example, comprising administering to a subject an antihypertensive and / or collagen modifying agent (referred to herein as “AHCM” or “AHCM agent”) under conditions of dosage of AHCM and an anti-cancer agent; Optionally administering a cancer treatment).

In one embodiment, the method includes one or more of the following:
(A) the subject is selected or identified as in need of AHCM acceptance based on the need for improved delivery and / or efficacy of the treatment (eg, cancer treatment);
(B) an entity having a hydrodynamic diameter greater than about 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm, eg, nanoparticles As administered AHCM, therapy (eg, cancer therapy), or both;
(C) the subject has a history of treatment (or lack of treatment) for hypertension as described herein, eg, subject within 5 days of onset of cancer diagnosis or AHCM dosing, 10 Within a day, within 30 days, within 60 days, or within 100 days, either a dose (eg, a dose sufficient to substantially reduce a subject's blood pressure, or a sub-anti-hypertentive dose) A) AHCM, eg, AHCM listed herein or any AHCM. In one embodiment, the subject is not hypertensive or hypertensive prior to administration of AHCM;
(D) The subject is treated according to the dosing schedule described herein. For example, AHCM administration is started before the start of cancer treatment administration, e.g., at least 1 day, 2 days, 3 days, or 5 days before cancer treatment, or 1 week, 2 weeks, 3 weeks, 4 Started weekly, five weeks ago, or earlier (eg, AHCM is administered at least two weeks before cancer treatment);
(E) To provide AHCM and cancer treatment in accordance with the dosing schedule described herein. For example, after a first course of treatment with a non-hypertensive dose of AHCM is provided, a second high-dose course of treatment with AHCM is provided, for example at a dose that is greater than or equal to the standard antihypertensive dose (eg, the second course is Administered on a time course that would counter the pressor effects of anti-cancer treatment);
(F) at least 1 hour, 5 hours, 10 hours, or 24 hours; at least 2 days, 5 days, 10 days, or 14 days; at least 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks; AHCM is administered substantially continuously over a period of at least 1 year, 2 years, 3 years, 4 years, or 5 years;
(G) AHCM is administered sequentially and / or simultaneously with treatment, eg, cancer treatment. AHCM and treatment may be administered in any order (in the same or different dosages) and may overlap with the treatment. In one embodiment, AHCM is administered prior to treatment (eg, as described in step (d)). In other embodiments, AHCM is administered sequentially and concurrently with treatment (eg, AHCM is administered prior to treatment (eg, as described in step (d)) and administered concurrently with treatment. ) In yet other embodiments, the treatment is administered first and AHCM is administered after initiation of the treatment. In embodiments where administration of AHCM and treatment is simultaneous, administration of AHCM and treatment includes clinically appropriate, eg, (i) as a combination treatment, (ii) a period of treatment with either AHCM or treatment, Or (iii) may be continued as a combination of (i) and (ii) in any order.

  In one embodiment, AHCM is administered in an amount sufficient to alter (eg, enhance) the dispersion or efficacy of the treatment, eg, cancer treatment. In some embodiments, AHCM alone is insufficient to inhibit or prevent tumor growth, but to modify (eg, enhance) the dispersion or efficacy of a treatment, eg, cancer treatment. It is administered in a sufficient amount.

  In one embodiment, the AHCM is a decrease in collagen level or production, decreased tumor fibrosis, increased stromal tumor transport, improved tumor perfusion, or penetration of cancer therapeutics in the tumor or tumor vasculature in a subject. It is administered at a dose that causes one or more of the enhanced diffusion.

In one embodiment, AHCM is
Angiotensin II receptor blocker (AT ₁ blocker),
Renin angiotensin aldosterone antagonist ("RAAS antagonist"),
Angiotensin converting enzyme (ACE) inhibitor,
Thrombospondin 1 (TSP-1) inhibitor,
Transforming growth factor β1 (TGFβ1) inhibitor,
It is selected from one or more of a connective tissue growth factor (CTGF) inhibitor or a combination of two or more thereof.

  Unless the context states otherwise, the term “AHCM” can refer to one or more agents described herein.

  The method may comprise one, two, three, or more AHCMs alone or in combination with one or more cancer treatments.

  In one embodiment, the AHCM is a RAAS antagonist. In one embodiment, the RAAS antagonist is one of aliskiren (TEKTURNA®, RASILEZ®), remixylene (Ro 42-5892), enalkylene (A-64662), SPP635, or derivatives thereof, or Selected from multiple species.

In another embodiment, AHCM is an AT ₁ inhibitor. In one embodiment, the AT ₁ blocker is losartan (COZAAR®), candesartan (ATACAND®), eprosartan mesylate (TEVETEN®), EXP 3174, irbesartan (AVAPRO®) ), L158,809, olmesartan (BENICAR (registered trademark)), salaracin, telmisartan (MICARDIS (registered trademark)), valsartan (DIOVAN (registered trademark)), or a derivative thereof. .

  In yet another embodiment, the AHCM is an ACE inhibitor. In one embodiment, the ACE inhibitor is benazepril (LOTENSIN®), captopril (CAPOTEN®), enalapril (VASOTEC®), fosinopril (MONOPRIL®), lisinopril (PRINIVIL®). Trademark), ZESTRIL (registered trademark), moexipril (UNIVASC (registered trademark)), perindopril (ACEON (registered trademark)), quinapril (ACCUPRIL (registered trademark)), ramipril (ALTACE (registered trademark)), trandolapril ( MAVIK (registered trademark)), or one or more of them.

  In yet another embodiment, the AHCM is a TSP-1 inhibitor. In one embodiment, the TSP-1 inhibitor is selected from one or more of ABT-510, CVX-045, LSKL, or derivatives thereof.

  In one embodiment, the AHCM is a TGFβ1 inhibitor, such as an anti-TGFβ1 antibody, a TGFβ1 peptide inhibitor. In certain embodiments, the TGFβ1 inhibitor is CAT-192, Fresolimumab (GC 1008), LY2157299, Peptide 144 (P144), SB-431542, SD-208, US Patent No. 7,846,908 and US Patent Application Publication No. 2011 / [0008] It is selected from one or more of the compounds described in No. 364, or their derivatives.

  In yet another embodiment, AHCM is a CTGF inhibitor. In certain embodiments, the CTGF inhibitor is one or more of the compounds described in DN-9693, FG-3019, and European Patent Application Publication No. 1839655, US Pat. No. 7,622,454, or derivatives thereof. Selected from seeds.

  The exemplary AHCM described herein is not limiting, for example, derivatives of AHCM described herein may be used in the methods described herein.

AHMC Dosage and Dosage Form The method of the invention uses AHCM to enhance treatment (eg, cancer treatment).

In one embodiment, AHCM is administered at a dose corresponding to a standard medical dose. Standard medical doses of AHCM are available in the art. For example, when AHCM is the AT ₁ inhibitor, losartan, the standard medical dose used for antihypertensive in humans is about 25-100 mg / day. In the methods of the present invention, losartan can be administered orally, either alone or in combination with the cancer treatment described herein, on a daily schedule (once or twice daily). Losartan can be provided in about 12.5 mg, 25 mg, 50 mg, or 100 mg dosage forms (eg, oral tablets).

Exemplary standards for medical doses of other AT ₁ inhibitors used for antihypertensive or anti-heart failure in humans are as follows: 4-32 mg / day candesartan (ATACAND®) (eg Available in oral dosage forms containing 4 mg, 8 mg, 16 mg or 32 mg candesartan; 400-800 mg / day eprosartan mesylate (TEVETEN®) (eg 400 mg or 600 mg Available in dosage forms for oral administration containing a large amount of eprosartan); 150-300 mg / day irbesartan (AVAPRO®) (eg, for oral administration containing 150 mg or 300 mg irbesartan) 20-40 mg / day olmesartan (BENICAR®) (available in dosage forms for oral administration containing 5 mg, 20 mg, or 40 mg olmesartan); 20-80 mg / day Day of Telmisartan (MICA RDIS®) (eg available in dosage forms for oral administration containing 20 mg, 40 mg, or 80 mg telmisartan); and 80-320 mg / day valsartan (DIOVAN®) (eg Available in oral dosage forms containing 40 mg, 80 mg, 160 mg, or 320 mg valsartan).

  In one embodiment, AHCM is administered at a non-hypertensive dose (eg, a dose that has no significant effect on mean arterial blood pressure when administered to a hypertensive subject; or a dose that is less than a standard medical antihypertensive dose). In one aspect, the AHCM measures the mean arterial blood pressure of a subject, as measured, for example, at a steady state plasma level for a given dosage after administration of a preselected number of times at that dosage. Administered in an amount that does not substantially reduce. In one aspect, AHCM is less than 1%, less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40%, less than 45%, less than 50% , Administered at least once in a dose that reduces the mean arterial blood pressure in the subject. In one aspect, the AHCM is less than 1%, less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, 35% of the reduction caused by the standard medical antihypertensive dose for that AHCM Less than, less than 40%, less than 45%, less than 50%, less than 60%, less than 70%, less than 80%, less than 90%, administered at a dose that lowers blood pressure. In one embodiment, the AHCM causes the subject's blood pressure to be in the normal range, e.g., about 120 systolic and about 80 diastolic, or the subject's blood pressure is 120 +/- 5 systolic and 80+ / -5 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60% of the dose that would be in the diastolic range It is administered at doses less than 70%, 80%, 90%.

In one embodiment, the AHCM is a dose that is less than the standard medical dose used for antihypertensive or anti-heart failure (e.g., 0.01, 0.02, 0.03, 0.04, 0.05, Less than 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7). Standard medical doses of AHCM are available in the art. For example, if AHCM is the AT ₁ inhibitor, losartan and the standard medical dose is about 25-100 mg / day, the non-optimal antihypertensive dose is 0.25-17.5, 0.5-15, 1.3-12, 1.5-12, 2 It can be in the range of -12, 2-10, 2-5, 2-3 mg / day, typically 2 mg / day. In one embodiment, the AHCM is losartan and a dose less than 25 mg / day, 20 mg / day, 15 mg / day, 10 mg / day, 5 mg / day, 4 mg / day, 3 mg / day, 2 mg / day, 1 mg / day Is administered. Losartan can be administered orally alone or in combination with the cancer therapeutics described herein at a non-hypertensive dose of 2-3 mg / day on a daily schedule (once or twice daily) . Exemplary standards for medical doses for other AT ₁ inhibitors are as follows: 4-32 mg / day candesartan (ATACAND®), 400-800 mg / day eprosartan mesylate (TEVETEN (Registered trademark)), 150-300 mg / day irbesartan (AVAPRO (registered trademark)), 20-40 mg / day olmesartan (BENICAR (registered trademark)), 20-80 mg / day telmisartan (MICARDIS (registered trademark)) , And 80-320 mg / day valsartan (DIOVAN®). In one embodiment, AHCM a medical buck dose or medical anti heart failure doses of standard lower doses (e.g., candesartan, eprosartan, irbesartan, olmesartan, medical antihypertensive dose or for other AT ₁ inhibitors such as telmisartan, and valsartan The standard dose of medical anti-cardiac insufficiency is less than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7). In certain embodiments, the AHCM is a dosage form smaller than the standard of a medical antihypertensive dosage form or a medical anti-heart failure dosage form (eg, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 of a standard medical dosage form). 0.1, 0.15, 0.16, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7). For example, when AHCM is losartan, the dosage form is about 0.5 mg to 11 mg; 1 mg to 10 mg; 1 to 5 mg, or 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or 10 mg obtain. In some embodiments, losartan is less than 12.5 mg, e.g., about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg , About 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, or about 12 mg in dosage forms (eg, oral tablets) .

  In one embodiment, AHCM is formulated as a tablet (eg, an oral tablet). In other embodiments, AHCM is formulated for other routes of administration, such as subcutaneous or intravenous administration.

  In some embodiments, a non-hypertensive dose of AHCM, or a dose of AHCM that is less than the standard medical dose used for antihypertensive or anti-heart failure, inhibits tumor growth or progression when administered alone to a subject Insufficient dose to do or prevent.

  In yet another embodiment, the AHCM is greater than the standard medical dose used for antihypertensive or anti-heart failure (eg, 1.1 times, 1.5 times, 1.7 times the standard medical dose used for anti-hypertensive or anti-heart failure). Twice, twice, three times, four times, five times, more than 10 times or more doses). Standard medical doses of AHCM are available in the art; some of which are exemplified herein.

  In other embodiments, the AHCM is a dosage form larger than the standard for a medical antihypertensive dosage form or a medical anti-heart failure dosage form (eg, 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, more than 10 times or more dosage form). Standard medical dosage forms of AHCM are available in the art; some of which are exemplified herein.

  In some embodiments, does a dose of AHCM that is comparable to or greater than a standard of a medical antihypertensive dose or a medical anti-heart failure dose inhibit tumor growth or progression when administered alone to a subject? Or it may be a dose that is insufficient to prevent.

  In other embodiments, the anticancer agent results in a higher dosage or higher anticancer agent level compared to a reference, eg, dosage on a package insert, standard medical dosage, or maximum tolerated dose (MTD). Will be administered on a schedule.

  In certain embodiments, the anticancer agent is administered at a lower dosage or in a plan that results in a lower level of anticancer agent compared to a reference, e.g., a dosage on a package insert, a standard medical dosage, or MTD. The In some embodiments, the anticancer agent is not effective to inhibit or prevent tumor growth or progression when administered alone, but does it inhibit tumor growth or progression when administered in combination with AHCM? Or is administered in an amount sufficient to prevent.

  In some embodiments, the cancer therapy or cancer therapeutic agent, when administered in combination with AHCM, treats cancer in a subject at a dose that is less than the lowest dose that would be used in the absence of AHCM, or To prevent, it is administered to the subject.

  In some embodiments, when both an AHCM agent and a cancer therapy or cancer therapeutic agent are administered to a subject, the dose of hypotensive and / or collagen modifying agent is used to treat a hypertension-related disorder or heart failure. The dose of cancer therapy or cancer therapeutic agent may be less than the lowest dose that would be used in the absence of AHCM to treat or prevent cancer in the subject. It can be a dose.

  In some embodiments, the dose of cancer therapy or cancer therapeutic administered to the subject is less than the lowest dose that would be used alone to treat a patient with cancer and is administered to the subject as an adjuvant. The dose of AHCM agent may be less than the lowest dose that would be used alone to treat cancer. In such embodiments, the dose of AHCM agent administered to the subject as an adjuvant may be a non-hypertensive dose or is comparable to or comparable to a standard medical dose for the treatment of hypertension or heart failure. There may be more.

  In some embodiments, the dose of cancer therapy or cancer therapeutic administered to the subject is less than the lowest dose that would be used in the absence of AHCM to treat patients with cancer, and the subject as an adjuvant The dose of AHCM agent administered to the patient is less than the lowest dose that would be used alone to treat cancer, but is sufficient to improve the efficacy of cancer therapy or the delivery of cancer therapeutics to the tumor Is. In such embodiments, the dose of AHCM agent administered to the subject as an adjuvant may be a non-hypertensive dose or is comparable to or comparable to a standard medical dose for the treatment of hypertension or heart failure. There may be more.

  Methods for determining the lowest dose of agents for treatment, eg, anticancer agents and / or AHCM are well known to those skilled in the art. For example, a skilled artisan can treat in an animal model corresponding to a particular type of cancer, for example, by administering different doses of AHCM and / or anticancer agents to an animal and monitoring tumor growth relative to a control. The lowest dose of AHCM and / or anticancer agent effective for the purpose can be determined. The control can be an animal treated with an anticancer agent alone (ie, in the absence of AHCM).

  AHCM and treatment (eg, cancer treatment) can be administered in combination, eg, sequentially and / or simultaneously, as described herein. AHCM and treatment may be administered in any order (in the same or different dosages) and / or may overlap with the treatment. In one embodiment, AHCM is administered prior to treatment. In other embodiments, AHCM is administered sequentially and concurrently with the treatment (eg, AHCM is administered prior to treatment and administered concurrently with treatment). In yet other embodiments, the cancer treatment is administered first and AHCM is administered after initiation of the cancer treatment. In embodiments where administration of AHCM and treatment is simultaneous, administration of AHCM and cancer treatment includes clinically appropriate (i) combination therapy, (ii) a period of treatment with either AHCM or cancer treatment, or (Iii) may be continued as a combination of (i) and (ii) in any order.

  Administration of AHCM can be substantially continuous. For example, administration of AHCM can be substantially continuous over a period of at least 1 hour, 5 hours, 10 hours, 24 hours; 2 days, 5 days, 10 days, 14 days, or more.

  In other embodiments, administration of AHCM may be intermittent, eg, having gaps at predetermined intervals during the course of treatment. In certain embodiments, two or more doses of AHCM are administered alone or in combination with a treatment (eg, cancer treatment). In one embodiment, AHCM is administered at a non-optimal antihypertensive and antihypertensive dose during the course of treatment. For example, before or at the time of therapy (eg, cancer therapy) (eg, treatment with an anti-cancer agent that increases mean arterial blood pressure, eg, treatment with an anti-angiogenic agent (eg, Avastin, sunitinib, or sorafenib)) An optimal antihypertensive dose of AHCM can be administered; then a second antihypertensive dose of AHCM can be administered.

The size of the therapeutic entity The methods described herein allow for increased flexibility in the range of treatment modalities used or selected, eg, the size of the therapeutic entity. Thus, in one embodiment, AHCM is administered as an entity having a hydrodynamic diameter greater than about 1 nm, 5 nm, 10 nm, 100 nm, 500 nm, or 1,000 nm. For example, the AHCM can be a protein, such as an antibody. AHCM may be administered as nanoparticles, eg, polymer nanoparticles or liposomes, including small molecule therapeutics or proteins, eg, AHCM as an antibody.

  In one embodiment, the cancer treatment is a cancer treatment (also referred to herein as an “anticancer agent”) or the second treatment is about 1 nm, 5 nm, 10 nm, 20 nm, 50 nm, 75 nm, 100 nm, Administered as an entity having a hydrodynamic diameter greater than 150 nm, 200 nm, 500 nm, or 1,000 nm. For example, the anticancer agent can be a protein, such as an antibody. The anticancer agent may be administered as a nanoparticle, eg, a polymer nanoparticle or liposome, comprising a small molecule therapeutic agent (ie, a molecule having a hydrodynamic diameter of about 1 nm or less) or a protein, eg, an anticancer agent as an antibody. .

  In one embodiment, the AHCM is administered as an entity having a hydrodynamic diameter greater than about 1 nm (eg, greater than about 1 nm, 5 nm, 10 nm, 20 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 500 nm, or 1,000 nm). And the anti-cancer agent is administered as an entity having a hydrodynamic diameter of about 1 nm or less. In one embodiment, the AHCM is present in an entity that does not include a chemotherapeutic agent. AHCM may be formulated into a long release formulation for extended release, for example, a substantially continuous release over hours, days, weeks, months, or years.

  In one aspect, AHCM is administered as an entity having a hydrodynamic diameter of about 1 nm or less and the anticancer agent is greater than about 1 nm (eg, about 1 nm, 5 nm, 10 nm, 20 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm Administered as an entity having a hydrodynamic diameter (greater than 500 nm, or 1,000 nm).

  In one embodiment, AHCM is administered as an entity having a hydrodynamic diameter of about 1 nm or less and the anticancer agent is administered as an entity having a hydrodynamic diameter of less than about 1 nm.

  In one embodiment, the AHCM is administered as an entity having a hydrodynamic diameter greater than about 1 nm (eg, greater than about 1 nm, 5 nm, 10 nm, 20 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 500 nm, or 1,000 nm). And the anti-cancer agent is administered as an entity having a hydrodynamic diameter greater than about 1 nm (eg, greater than about 1 nm, 5 nm, 10 nm, 20 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 500 nm, or 1,000 nm) . The AHCM and the anticancer agent may be in separate entities or in the same entity. For example, when provided as separate entities, AHCM can be provided as a first nanoparticle and an anti-cancer agent can be provided as a second nanoparticle (eg, the second nanoparticle is combined with the first nanoparticle). With different structural properties (eg size or composition) or functional properties (eg release kinetics or pharmacodynamic properties). Alternatively, the AHCM and anticancer agent may be provided in the same entity, eg, in the same nanoparticle.

  In one embodiment, AHCM is 1 nm or less or 2 nm or less; 2-20 nm, 10-25 nm, 20-40 nm, 40 nm, 50-150 nm; 10-100 nm, 15-100 nm, 20-100 nm, 25-100 nm, 35-100 nm 40-100 nm, 45-100 nm, 50-100 nm; 10-200 nm, 15-200 nm, 20-200 nm, 25-200 nm, 35-200 nm, 40-200 nm, 45-200 nm, 50-200 nm; 10-500 nm, 15 -500 nm, 20-500 nm, 25-500 nm, 35-500 nm, 40-500 nm, 45-500 nm, 50-500 nm, 75-500 nm, 100-500 nm, 150-500 nm, 200-500 nm, 300-500 nm; and 10- 1000 nm, 15-1000 nm, 20-1000 nm, 25-1000 nm, 35-1000 nm, 40-1000 nm, 45-1000 nm, 50-1000 nm, 75-1000 nm, 100-1000 nm, 150-1000 nm, 200-1000 nm, 300-1000 nm; Or selected from therapeutic entities having a hydrodynamic diameter of 10 nm, 15 nm, 20 nm, 25 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, or 200 nm.

  In one embodiment, the AHCM is a small molecule therapeutic agent; is a protein, eg, an antibody; or is provided in nanoparticles.

  In one embodiment, the anticancer agent or second therapeutic agent is 1 nm or less or 2 nm or less; 2-20 nm, 10-25 nm, 20-40 nm, 40 nm, 50-150 nm; 10-100 nm, 15-100 nm, 20-100 nm, 25 -100 nm, 35-100 nm, 40-100 nm, 45-100 nm, 50-100 nm; 10-200 nm, 15-200 nm, 20-200 nm, 25-200 nm, 35-200 nm, 40-200 nm, 45-200 nm, 50-200 nm 10-500 nm, 15-500 nm, 20-500 nm, 25-500 nm, 35-500 nm, 40-500 nm, 45-500 nm, 50-500 nm, 75-500 nm, 100-500 nm, 150-500 nm, 200-500 nm, 300 ~ 500nm; and 10 ~ 1000nm, 15 ~ 1000nm, 20 ~ 100nm, 25 ~ 1000nm, 35 ~ 1000nm, 40 ~ 100nm, 45 ~ 1000nm, 50 ~ 1000nm, 75 ~ 1000nm, 100 ~ 1000nm, 150 ~ 1000nm, 200 ~ Or selected from therapeutic entities having a hydrodynamic diameter of 10 nm, 15 nm, 20 nm, 25 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, or 200 nm.

  In one embodiment, the anti-cancer agent is a small molecule therapeutic with a hydrodynamic diameter of 1 nm or less; a protein, eg, an antibody; or provided in nanoparticles.

  In one embodiment, the AHCM or anticancer agent or second therapeutic agent can each independently be provided as an entity having the following size range (nm): a hydrodynamic diameter of 1 nm or less or 0.1-1.0 nm, eg, typical Small molecules; hydrodynamic diameters of 5-20 nm or 5-15 nm, eg of proteins, eg antibodies; or 10-5,000 nm, 20-1,000 nm, 10-500 nm, 10-200 nm, 10 -150 nm, or 10-100 nm, 10-25 nm, 20-40 nm, 40 nm, 50-150 nm; 10-100 nm, 15-100 nm, 20-100 nm, 25-100 nm, 35-100 nm, 40-100 nm, 45-100 nm, 50 to 100 nm; 10 to 200 nm, 15 to 200 nm, 20 to 200 nm, 25 to 200 nm, 35 to 200 nm, 40 to 200 nm, 45 to 200 nm, 50 nm to 200 nm; 10 to 500 nm, 15 to 500 nm, 20 to 500 nm, 25 to 500 nm, 35-500 nm, 40-500 nm, 45-500 nm, 50-500 nm, 75-500 nm, 100-500 nm, 150-500 nm, 200-500 nm, 300-500 nm; and 10-1000 nm, 15-1000 nm, 20-1000 nm 25-1000nm, 35-1000nm, 40 ~ 1000nm, 45-1000nm, 50-1000nm, 75-1000nm, 100-1000nm, 150-1000nm, 200-1000nm, 300nm-1000nm; or 10nm, 15nm, 20nm, 25nm, 35nm, 45nm, 50nm, 75nm, 100nm, Hydrodynamic diameter of 150 nm or 200 nm, for example typical nanoparticle range.

Subjects The methods described herein can be used to treat a subject having the characteristics or needs defined herein. In embodiments, the subject or treatment for the subject is selected based on the characteristics described herein. In one embodiment, the methods described herein allow for optimized selection of patients and treatments.

  In some embodiments, a subject can be selected or identified before being subjected to any aspect of the methods described herein.

  In one embodiment, the subject is selected as in need of AHCM acceptance based on treatment optimization, e.g., based on the need for improved delivery and / or efficacy of treatment (e.g., cancer treatment) or Identified.

  In one embodiment, the subject has no hypertension or has not been treated for hypertension at the beginning of AHCM treatment or at the time of patient selection for administration of AHCM.

  In one aspect, the subject, e.g., patient, has an AHCM, e.g., herein, within 5 days, within 10 days, within 30 days, within 60 days, or within 100 days of diagnosis of cancer or initiation of AHCM dosing. Not receiving the listed AHCM or any AHCM dose.

  In one aspect, a subject, eg, a subject with normal or low blood pressure, is selected or identified based on the need for AHCM, eg, based on treatment optimization, eg, treatment (eg, cancer treatment). Based on the need for improved delivery and / or efficacy of AHCM, it is selected or identified as requiring the reception of AHCM.

  In some embodiments, a subject in need of receiving AHCM, based on improved delivery or efficacy of cancer treatment or need for treatment optimization, partially responds to a single cancer treatment? Or it is a subject that does not respond.

  In one embodiment, AHCM is selected for treatment of a subject based on the ability to optimize cancer treatment, eg, the ability to improve the delivery and / or efficacy of cancer therapy.

  In one aspect, the subject being treated is not a hypertensive patient, for example, has no history of hypertension or has not been treated with an antihypertensive agent. In one embodiment, the subject being treated has normal or low mean arterial blood pressure. In other embodiments, the subject to be treated has never received antihypertensive therapy or has not been treated with antihypertensive therapy.

  In one embodiment, the subject is in need of cancer treatment. In another embodiment, the subject is in need of or is under anticancer therapy (eg, treatment with any of the anticancer therapeutics described herein). In certain embodiments, the method comprises determining whether the subject has cancer (eg, solid cancer or fibrotic cancer) and, in response, administering AHCM and an anticancer agent.

  In other embodiments, the subject is at risk of developing cancer or having recurrence, such as a subject with a preneoplastic or genetic predisposition to cancer (eg, a subject with a BRCA1 mutation; or in an adjuvant environment Breast cancer patients being treated (eg, with tamoxifen).

  In other embodiments, the subject has early stage cancer or more advanced (eg, moderate) or metastatic cancer.

  In one embodiment, the subject has pancreatic cancer (eg, pancreatic adenocarcinoma), breast cancer, colorectal cancer, lung cancer (eg, small cell lung cancer or non-small cell lung cancer), skin cancer, ovarian cancer, prostate cancer, cervical cancer. Having a solid fibrotic tumor selected from one or more of gastrointestinal cancer (eg, carcinoid or matrix), gastric cancer, head and neck cancer, kidney cancer, or liver cancer, or metastatic lesions thereof. In other embodiments, the subject has a hyperproliferative cancer condition (eg, benign, pre-malignant, or malignant condition). A subject can be a person at risk of having a disorder, for example, a subject having a relative suffering from a disorder, or a subject having a genetic trait associated with the risk of disorder. In one embodiment, the subject may be symptomatic or asymptomatic. In one embodiment, the subject retains an oncogenic gene or oncogenic gene product modification. In one aspect, the subject is a patient undergoing cancer treatment (eg, the same or other anticancer agent, surgery, and / or radiation). In one aspect, the subject is a patient who has undergone cancer treatment (eg, other anticancer agents, surgery, and / or radiation). In one embodiment, the subject has never been treated with cancer therapy.

  In one embodiment, the subject has a metastatic cancer, such as a cancer disclosed herein (pancreatic cancer (eg, pancreatic adenocarcinoma), breast cancer, colorectal cancer, lung cancer (eg, small cell lung cancer or non-small cell). A patient with a metastatic form of lung cancer), skin cancer, ovarian cancer, or liver cancer.

  In one embodiment, the subject is a patient with a cancer or hyperproliferative disorder that is resistant to treatment.

  In some embodiments, the subject selected to be subjected to the methods or pharmaceutical compositions herein does not have kidney disease or kidney related disease.

  In one embodiment, the subject to be treated is a mammal, eg, a primate, typically a human (eg, a patient having or at risk of having a cancer or tumor described herein). .

  In one embodiment, the subject being treated has a hyperproliferative disorder, eg, a hyperproliferative connective tissue disorder (eg, a hyperproliferative fibrotic disease). In one embodiment, the hyperproliferative fibrotic disease is systemic or organ specific. Exemplary hyperproliferative fibrotic diseases include systemic disorders (eg, systemic sclerosis, multifocal fibrosis, scleroderma graft-versus-host disease in bone marrow transplant recipients, renal systemic fibrosis) , Scleroderma), and organ-specific disorders such as, but not limited to, lung, liver, heart, kidney, pancreas, skin, and other organ fibrosis.

  In other embodiments, the subject to be treated has a hyperproliferative hereditary disorder, eg, a hyperproliferative hereditary disorder selected from Marfan syndrome or Royce Dietz syndrome.

  In other embodiments, the hyperproliferative disorder (eg, hyperproliferative fibrotic disorder) is chronic obstructive pulmonary disease, asthma, aortic aneurysm, radiation-induced fibrosis, skeletal muscle myopathy, diabetic nephropathy, and / or arthritis It is selected from one or more of them.

Combination Therapy In one embodiment, AHCM is administered in combination with a therapy, eg, a cancer therapy (eg, one or more of anti-cancer drugs, surgery, and / or radiation). The terms “chemotherapeutic agent”, “chemotherapeutic agent”, and “anticancer agent” are used interchangeably herein. Administration of AHCM and cancer treatment may be sequential (with or without overlap) or simultaneous. Administration of AHCM may be continuous or intermittent throughout the course of treatment (eg, cancer treatment).

  In one embodiment, AHCM administration is initiated prior to initiation of administration of cancer treatment, e.g., at least 1 day, 2 days, 3 days, or 5 days prior to cancer treatment, or 1 week, 2 weeks, 3 weeks. Starts before, 4 weeks, 5 weeks, or earlier (eg, AHCM is administered at least 2 weeks before cancer treatment). In one embodiment, it is initiated only 5 days, 10 days, 20 days, 30 days, 60 days, or 120 days before the start of cancer treatment. In one aspect, AHCM administration is initiated prior to cancer treatment and until criteria, e.g., time-based criteria, e.g., a predetermined number of days of AHCM administration or a predetermined number of AHCM administrations are met. Cancer treatment is not started. In one embodiment, the criteria are those that meet a preselected level of AHCM, eg, a preselected level in serum or plasma. In one embodiment, the criteria include collagen I, collagen III, collagen IV, transforming growth factor β1 (TGFβ1), connective tissue growth factor (CTGF), or thrombospondin 1 (TSP-1). Meet preselected plasma or serum levels of non-limiting biomarkers. In another embodiment, the criteria are those that meet a preselected level of modification of tumor morphology.

  In one embodiment, administration of AHCM is sequential and / or simultaneous with the treatments described herein, eg, cancer treatment.

  In one embodiment, AHCM is administered or a preselected level of AHCM, eg, plasma level of AHCM, is maintained during a preselected portion of the time a subject receives cancer treatment. By way of example, AHCM treatment is maintained for the entire period for which the cancer treatment is administered, or for the entire period for which the preselected level of anticancer agent persists in the subject.

  Typically, treatment with AHCM continues throughout the cancer treatment schedule. In yet other embodiments, administration of AHCM is interrupted prior to termination of cancer treatment. In other embodiments, administration of AHCM is continued after completion of cancer treatment.

  In one embodiment, two or more doses of AHCM are administered alone or in combination with cancer treatment. In one embodiment, AHCM is administered at a non-hypertensive dose and a hypotensive dose during the course of treatment. For example, a non-optimal antihypertensive dose of AHCM prior to or at the time of cancer therapy (eg, treatment with an anti-cancer agent that increases mean arterial blood pressure, eg, treatment with an anti-angiogenic agent (eg, Avastin, sunitinib, or sorafenib)) Then a hypotensive dose of AHCM can be administered.

  In one embodiment, the AHCM (single or combined) has a period of at least 15 minutes, 30 minutes, 45 minutes; a period of at least 1 hour, 5 hours, 10 hours, 24 hours; at least 2 days, 5 days 10 days, 14 days; at least 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks; at least 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 Monthly, 9 months, 10 months, 11 months; administered substantially continuously over a period of at least 1 year, 2 years, 3 years, 4 years, 5 years, or longer. In one embodiment, AHCM is administered as a sustained release formulation. In certain embodiments, AHCM is formulated for continuous delivery, eg, oral delivery, subcutaneous delivery, or intravenous continuous delivery. In one embodiment, AHCM is administered via an implantable device, such as a pump (eg, a subcutaneous pump), an implant, or a depot. The method of delivery is such that the AHCM dose described herein (eg, a standard dose, a non-hypertensive dose, or a dose greater than the standard dose) is a predetermined period of time (eg, at least 15 minutes, 30 minutes, 45 minutes; 1 hour, 5 hours, 10 hours, 24 hours, 2 days, 5 days, 10 days, 14 days; 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks; 2 months, 3 months, 4 months , 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months; periods of 1 year, 2 years, 3 years, 4 years, 5 years, or longer) and / or It can be optimized to be maintained in the subject. Substantially continuous or extended release AHCM delivery or formulation prevents cancer over a period of hours, days, weeks, months, or years (with or without chemotherapy) Or it may be used for treatment. In one embodiment, the cancer treatment comprises nanotherapy (eg, viral cancer therapeutic agent (eg, oncolytic herpes simplex virus (HSV)), lipid nanoparticle (eg, liposomal formulation (eg, PEGylated liposomal doxorubicin (DOXIL®) Trademark)))), or polymer nanoparticles); antibodies that bind to cancer targets; RNAi or antisense RNA agents; chemotherapeutic agents (eg, cytotoxic or mitotic inhibitors); radiation; or surgery; Additional examples of anti-cancer treatments that can be used in combination with AHCM are provided below, selected from one or more of any combination.

  In other embodiments, AHCM and / or treatment (eg, treatment of cancer or hyperproliferation) is a profibrotic pathway (eg, a pathway that is dependent or independent of activation of TGFβ and / or CTGF). In combination with an inhibitor ("profibrotic pathway inhibitor"). In one embodiment, AHCM and / or cancer treatment is administered in combination with one or more of inhibitors of endothelin 1, PDGF, Wnt / β catenin, IGF-1, TNFα, and / or IL-4. Is done. In another embodiment, AHCM and / or cancer treatment is administered in combination with an inhibitor of endothelin 1 and / or PDGF. In other embodiments, AHCM and / or cancer treatment is chemokine receptor type 4 (CXCR4) (eg AMD3100, MSX-122); stroma-derived factor 1 (SDF-1) (eg tannic acid); hedgehog ( For example, GDC-0449, cyclopamine, or GANT58) is administered in combination with one or more inhibitors.

  Administration of AHCM, cancer treatment, and / or profibrotic pathway inhibitor may be sequential (eg, with or without overlap), or as described herein, or It may be simultaneous.

Cancer Therapy In one embodiment, the cancer being treated is an epithelial, mesenchymal, or hematological malignancy. In one aspect, the cancer to be treated is a solid tumor (eg, carcinoid, carcinoma, or sarcoma), soft tissue tumor (eg, heme malignancy), and metastatic lesions, eg, of the cancers disclosed herein. Any metastatic lesion. In one embodiment, the cancer being treated is a fibrotic or fibrogenic solid tumor, such as a tumor having one or more of limited tumor perfusion, compressed blood vessels, or fibrotic tumor stroma. It is. In one embodiment, the solid tumor is of pancreatic cancer (eg, pancreatic adenocarcinoma), breast cancer, colorectal cancer, lung cancer (eg, small cell lung cancer or non-small cell lung cancer), skin cancer, ovarian cancer, or liver cancer. Selected from one or more of the following: Additional examples of cancers to be treated are described herein below.

  In another embodiment, AHCM is administered in combination with a cancer treatment (eg, one or more of anti-cancer agents, surgery, and / or radiation). In one embodiment, the cancer treatment includes, for example, nanotherapy (eg, viral cancer therapeutics (eg, oncolytic herpes simplex virus (HSV)), lipid nanoparticles (eg, liposomal formulations (eg, PEGylated liposomal doxorubicin (DOXIL)). (Registered trademark))), or one or more types of nanotherapeutic agents comprising polymer nanoparticles); one or more types of cancer therapeutic antibodies (eg, anti-HER2 antibodies, anti-EGFR antibodies, Anti-CD20 antibodies); RNAi agents and antisense RNA agents; one or more chemotherapeutic agents (eg, low molecular weight chemotherapeutic agents including cytotoxic or mitotic inhibitors); radiation; or surgery, or thereof Including one or more of any combination, one or more AHCM and one or more therapeutic modalities (eg, first, second, third), nanocure Any combination of therapeutic agents, antibody agents, low molecular weight chemotherapeutic agents, radiation may be used, exemplary cancer therapeutic agents include nanotherapeutic agents (eg, one or more lipid nanoparticles (eg, liposomes) A formulation (eg, PEGylated liposomal doxorubicin (DOXIL®) or liposomal paclitaxel (eg, Abraxane®)), or one or more low molecular weight chemotherapeutic agents (eg, gemcitabine) , Cisplatin, epirubicin, 5-fluorouracil, paclitaxel, oxaliplatin, or leucovorin); one or more antibodies against a cancer target (eg, a growth factor receptor such as HER-2 / neu, HER3, VEGF); , Sunitinib, erlotinib, gefitinib, sorafenib, icotinib, lapatinib, neratinib One or more tyrosine kinase inhibitors, including low molecular weight drugs and antibody drugs, such as vandetanib, BIBW 2992 or XL-647, anti-EGFR antibodies (eg, cetuximab, panitumumab, saltumumab, nimotuzumab, nesitumumab, or matuzumab) Including, but not limited to, additional examples of chemotherapeutic agents used in combination therapy are described below.

  In one embodiment, the chemotherapeutic agent used in combination with AHCM is a cytotoxic agent or a cell division inhibitor. Exemplary cytotoxic agents include microtubule inhibitors, topoisomerase inhibitors (eg, irinotecan), or taxanes (eg, docetaxel), antimetabolites, mitotic inhibitors, alkylating agents, intercalating agents, signaling Agents that can interfere with the pathway, agents that promote apoptosis, and radiation are included. In yet other embodiments, the method comprises an immunomodulator, such as an immune cell such as IL-1, IL-2, IL-4, IL-6, IL-12, interferon alpha, interferon gamma, or GM-CSF. Can be used in combination with growth factors.

  In one embodiment, AHCM alone or in combination with one or more cancer treatments described herein for cancer prevention (eg, alone or in combination with a cancer prevention agent) It is administered during periods of active disorder or during periods of remission or less active disorder. AHCM alone or in combination with one or more cancer therapies described herein for cancer prevention, before treatment or prevention, simultaneously with treatment or prevention, after treatment or prevention Or it can be administered during remission of the disorder. In one embodiment, the cancer treatment is administered concurrently with AHCM, sequentially, or a combination of both.

  In one embodiment, AHCM is, for example, cancer in a high-risk subject (eg, a subject with a pre-neoplastic or cancer genetic predisposition (eg, a subject with a BRCA1 mutation); or a breast cancer patient treated with tamoxifen). Is administered alone or in combination with a cancer inhibitor.

  In some embodiments, AHCM, alone or in combination with cancer therapy, is a primary treatment for cancer, for example, previously administered another drug intended to treat cancer Used in subjects that never happen.

  In other embodiments, the AHCM alone or in combination with a cancer therapy is a secondary treatment for cancer, for example, previously administered another drug intended to treat cancer Used in some subjects.

  In other embodiments, the AHCM, alone or in combination with cancer therapy, is a tertiary, quaternary, or later treatment for cancer, e.g., two types aimed at treating cancer, 3 Used in subjects who have previously been administered a species, or more other drugs.

  In other embodiments, AHCM is administered as an adjunct therapy, eg, a treatment added to the main therapy.

  In one embodiment, AHCM is administered as an adjuvant treatment.

  In other embodiments, AHCM is administered as a neoadjuvant treatment.

  In some embodiments, AHCM is administered to a subject before or after surgical resection / removal of cancer.

  In some embodiments, AHCM is administered to a subject before, during, and / or after cancer radiotherapy.

  In some embodiments, the AHCM is a subject, eg, a cancer patient who will receive a cancer therapy (eg, treatment with a chemotherapeutic agent, radiation therapy, and / or surgery), a cancer patient who has received, or has received It is administered to some cancer patients.

  In other embodiments, AHCM is administered prior to cancer treatment. In other embodiments, AHCM is administered concurrently with cancer treatment. In yet other embodiments, the AHCM is administered prior to the cancer treatment and is administered concurrently with the cancer treatment. In the case of co-administration, AHCM may continue to be administered after completion of cancer treatment.

  In other embodiments, AHCM is administered sequentially with cancer treatment. For example, AHCM can be administered before initiating treatment with cancer therapy or after completing treatment with cancer therapy. In one embodiment, administration of AHCM overlaps with cancer treatment and continues after completion of cancer treatment. In one embodiment, AHCM is administered at the same time, sequentially, or as a combination where simultaneous administration is followed by either cancer treatment or monotherapy with either AHCM.

  In one embodiment, the method includes administering AHCM as the first therapeutic agent followed by administering a cancer treatment (eg, a second therapeutic agent, radiation therapy, and / or surgery). In another embodiment, the method comprises first administering a cancer treatment (eg, a first therapeutic agent, radiation therapy, and / or surgery), followed by administering AHCM as a second therapeutic agent. . In yet other embodiments, the method comprises administering AHCM in combination with a second, third, or more additional therapeutic agent (eg, an anticancer agent described herein).

  AHCM and / or the anticancer agents described herein can be systemically administered to a subject (e.g., oral, parenteral, subcutaneous, intravenous, rectal, intramuscular, intraperitoneal, intranasal, transdermal, or inhalation, Or intracavitary introduction). Typically, AHCM is administered orally. In certain embodiments, the AHCM and / or anticancer agent is administered intratumor (eg, via an oncolytic virus).

  In some embodiments, AHCM is administered as a pharmaceutical composition comprising one or more AHCM and a pharmaceutically acceptable excipient.

  In one embodiment, AHCM is administered in or present in a composition, eg, a pharmaceutical composition (eg, the same nanoparticle composition).

  In other embodiments, AHCM and cancer treatment are administered as separate compositions, eg, pharmaceutical compositions (eg, nanoparticle compositions). In other embodiments, AHCM and cancer treatment are administered separately, but administered via the same route (eg, either orally or both intravenously). In some embodiments, AHCM and cancer treatment are administered by different routes (eg, AHCM is administered orally and the cancer therapeutic is administered intravenously). In yet other examples, AHCM and cancer treatment are administered in the same composition, eg, a pharmaceutical composition.

Subject Monitoring The method of the present invention comprises, for example, one or more of tumor size; transforming growth factor β1 (TGFβ1), connective tissue growth factor (CTGF), or thrombospondin 1 (TSP-1). Level or signaling; tumor collagen I level; fiber content, interstitial pressure; biomarkers in plasma or serum, eg, collagen I, collagen III, collagen IV, TGFβ1, CTGF, TSP-1; one or more The level of cancer markers; the appearance of new lesions, metabolism, the rate of hypoxia; the appearance of new disease-related symptoms; the size of tissue mass, eg, reduction or stabilization; the quality of life, eg, the amount of disease-related pain Histological analysis, leaflet patterns, and / or presence of mitotic cells; tumor invasiveness, primary tumor angiogenesis, metastatic spread; multidisciplinary imaging techniques Further comprising monitoring the subject for one or more changes (eg, increase or decrease) of tumor size and location that can be visualized using; or any other parameter related to clinical outcome obtain. Subjects can be monitored in one or more of the following periods: before the start of treatment; during the treatment; or after one or more elements of treatment have been administered. Monitoring can be used to assess the need for further treatment with the same AHCM, alone or in combination with the same anticancer agent, or additional treatment with additional agents. In general, a decrease in one or more of the above parameters indicates an improved state of the subject.

  The methods of the invention can further comprise analyzing a nucleic acid or protein from the subject, eg, analyzing the genotype of the subject. The analysis can be performed, for example, to assess the particular dosage, delivery mode, delivery time, inclusion of an adjunct therapy, for example, the suitability of administration in combination with a second agent, or with another treatment. It can be used to select between, or generally to determine a potential drug response phenotype or genotype of a subject. The nucleic acid or protein can be analyzed at any stage of treatment, but preferably the appropriate dosage and treatment regimen of AHCM for the prophylactic or therapeutic treatment of the subject (e.g., the amount per treatment or To determine the frequency of treatment) before administration of AHCM and / or anticancer agents.

Dosage Form In another aspect, the invention features a pharmaceutically acceptable composition comprising AHCM and an anticancer agent, eg, a small molecule or protein, eg, an antibody, in a single dosage form. . In another embodiment, one or both of AHCM and the anticancer agent are provided in nanoparticles. The AHCM and the anticancer agent may be in separate entities or in the same entity. For example, when provided as separate entities, AHCM can be provided as a first nanoparticle and an anti-cancer agent can be provided as a second nanoparticle (eg, the second nanoparticle is combined with the first nanoparticle). With different structural properties (eg size or composition) or functional properties (eg release kinetics or pharmacodynamic properties). Alternatively, the AHCM and anticancer agent may be provided in the same entity, eg, in the same nanoparticle.

  In another aspect, the invention features a pharmaceutically acceptable composition (eg, a nanoparticle) comprising AHCM, eg, an AHCM described herein. In one embodiment, AHCM is present in a dosage larger than the dosage described herein, eg, a standard medical dosage form, a non-hypertensive dosage form, or a standard medical dosage form.

  In one embodiment, the AHCM is formulated in a dosage form according to a standard of a medical antihypertensive dosage form or a medical anti-heart failure dosage form, eg, a standard medical dosage form described herein.

  In certain embodiments, the AHCM is a dosage form smaller than the standard for a medical antihypertensive dosage form or a medical anti-heart failure dosage form (e.g., a standard medical dosage form, e.g., 0.01 times the standard medical dosage form described herein, 0.02 times, 0.03 times, 0.04 times, 0.05 times, 0.06 times, 0.07 times, 0.08 times, 0.09 times, 0.1 times, 0.15 times, 0.16 times, 0.2 times, 0.3 times, 0.4 times, 0.5 times, 0.6 times, 0.7 times Smaller formulation). In other embodiments, the AHCM is larger than the standard for a medical antihypertensive dosage form or a medical anti-heart failure dosage form (eg, a standard medical dosage form, eg, 1.1 times the standard medical dosage form described herein, 1.5 times Twice, 1.7 times, 2 times, 3 times, 4 times, 5 times, more than 10 times or more).

  In another aspect, the present invention provides a pharmaceutical composition comprising an anti-cancer agent, eg, an anti-cancer agent described herein, as a nano-particle, eg, a nanoparticle constructed for the methods described herein. Features an acceptable composition.

  In another aspect, the present invention alone or in combination with a therapy described herein, eg, an anti-cancer agent, includes AHCM, eg, a therapy optionally comprising instructions for the treatment of cancer. Features a kit. In one embodiment, the kit comprises one or more dosage forms of a pharmaceutical preparation or nanoparticle described herein.

In another aspect, the present invention optimizes the access of an agent, eg, a systemically administered agent, eg, a diagnostic or imaging agent, to a target tissue, eg, cancer, or a target tissue, For example, it features a method that optimizes delivery to cancer. The method includes administering to the subject an antihypertensive and / or collagen modifying agent (“AHCM”); and optionally comprising administering an agent, eg, a diagnostic or imaging agent, to the subject.

In one embodiment, the method includes one or more of the following:
(A) AHCM is an antihypertensive and is administered at a standard medical dose, a non-hypertensive dose, or a dose greater than a standard antihypertensive dose;
(B) the agent, for example a diagnostic or imaging agent, has a hydrodynamic diameter greater than 1 nm, 5 nm, or 20 nm, for example nanoparticles;
(C) the agent is an imaging agent, eg, a radiation agent, NMRA agent, contrast agent; or (d) the subject is treated with a dosing schedule described herein, eg, AHCM administration Before administration of the agent, for example, at least 1 day, 2 days, 3 days, or 5 days before administration of the agent, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or Be started before.

  In one embodiment, AHCM is administered in an amount sufficient to alter (eg, enhance) the dispersion or efficacy of the agent. In one embodiment, AHCM is sufficient to modify (eg, enhance) the dispersion or efficacy of the agent, but alone in an amount insufficient to inhibit or prevent tumor growth or progression. Administered.

  In one aspect, the AHCM is a decrease in collagen level or production, decreased tumor fibrosis, increased stromal tumor transport, improved tumor perfusion, or penetration or spread of a cancer therapeutic in a tumor or tumor vasculature in a subject. At a dose that causes one or more of the enhancements.

  In one aspect, the subject is further treated with a cancer therapy, eg, a therapy described herein.

  In one aspect, the subject is a human or non-human animal, such as a mouse, rat, non-human primate, horse, or cow.

  In another embodiment, the present invention includes AHCM, alone or in combination with an agent described herein, e.g., a diagnostic or imaging agent, e.g., instructions for the diagnosis of cancer. Features an optional diagnostic kit.

Screening Assay In another embodiment, the invention features a method or assay for identifying AHCM. The method or assay comprises preparing a cancer or cancer-related cell (eg, a culture of cancer-related fibroblasts); contacting the cancer or cancer-related cell with a candidate agent; in the presence or absence of the candidate agent. Detecting a change in the cancer cell. In one embodiment, the detected change comprises one or more of an increase or decrease in TGFβ1 level, connective tissue growth factor (CTGF) level, or collagen (eg, collagen 1) level. In one embodiment, the candidate agent is a renin angiotensin aldosterone antagonist (“RAAS antagonist”), an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (AT ₁ blocker), thrombospondin 1 (TSP). -1) It is selected from one or more of an inhibitor, a transforming growth factor β1 (TGFβ1) inhibitor, or a connective tissue growth factor (CTGF) inhibitor. Suitable candidate agents reduce one or more of TGFβ1 levels (eg, total TGFβ1 and / or active TGFβ1), connective tissue growth factor (CTGF) levels, or collagen levels.

  The method or assay is the same as the method or assay in which the treatment was performed, in the absence of a reference value, eg, a candidate agent, or a control agent, eg, positive agent (eg, losartan) or negative agent (eg, saline control). And comparing the difference between the treated method and the control method.

  The method or assay can be performed in vitro, in vivo, or a combination of both. In one embodiment, the method or assay comprises evaluating a candidate agent in vitro, eg, using a culture of cancer associated cells. In such embodiments, candidate agents are added to the culture medium; conditioned medium is analyzed for increases or decreases in TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen levels.

  In another embodiment, the candidate agent is administered to a subject, eg, an animal model, eg, an animal tumor model. In such embodiments, the candidate agent is administered to the subject under appropriate conditions; the subject is analyzed for increases or decreases in TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen levels. In one embodiment, the levels of these parameters are analyzed as described in the accompanying examples.

  In yet other embodiments, candidate agents evaluated using in vitro assays are tested in vivo.

  In another aspect, the invention provides a composition for the use of an AHCM agent alone or in combination with an anticancer agent described herein for the treatment of a cancer or tumor described herein. Or the use of an AHCM agent, alone or in combination with an anticancer agent described herein.

  Titles or numbered or lettered elements, such as (a), (b), (i), etc. are only presented for ease of reading. The use of title or numbered or lettered elements in this document does not require that the steps or elements be performed in alphabetical order or that the steps or elements must be distinguished from each other.

  All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

  Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

FIG. 6 is a bar graph panel showing the effect of losartan (10 μmol / L) on total TGFβ levels, active TGFβ levels, and collagen I synthesis in cancer-associated fibroblasts (CAF) in vitro. 2 shows the effect of losartan on collagen production in tumors. FIG. 2A shows collagen assessed by SHG imaging in HSTS26T tumors treated with losartan (10 mg / kg / day, 20 mg / kg / day, and 60 mg / kg / day) compared to controls over 2 weeks. Fig. 3 is a photographic panel showing a dose-dependent decrease in levels. Scale bar = 200 μm. FIG. 2B shows a dose-dependent decrease in collagen levels in tumors treated with losartan, 20 mg, 33%, and 67%, respectively, with 10 mg / kg / kg reduced SHG levels at the end of 15 days. Dose response curves of the effects of daily, 20 mg / kg / day, and 60 mg / kg / day losartan doses are shown. There was a statistically significant difference between the control group and the two higher doses (20 mg / kg / day and 60 mg / kg / day) ( ^* ). There was also a statistically significant difference between the 20 mg / kg / day group and the 60 mg / kg / day group (†). 2 is a bar graph showing the dose response of losartan to collagen content in HSTS26T tumors. Treatment with 20 mg / kg / day and 60 mg / kg / day of losartan resulted in a 42% and 63% reduction in collagen I staining, respectively. Staining in each treatment group was compared to a control group that received saline. 2 is a bar graph showing the effect of losartan in reducing mean arterial blood pressure (MABP) in mice in a dose-dependent manner. 20 mg / kg / day reduced MABP by 10 mmHg ( ^* p <0.04), but MABP remained within the normal range (70 mmHg to 95 mmHg) for SCID mice (13). Conversely, when animals were treated with 60 mg / kg / day, the 35 mm Hg ( ^** p <0.04) reduction in MABP was lower than the normal range MABP in SCID mice. Figure 2 shows the effect of losartan on collagen levels in tumors. FIG. 5A shows the results of collagen I and nuclear immunostaining in tumor sections in L3.6pl and MMTV tumors treated with control and losartan (20 mg / kg / day). Scale bar = 100 μm. Losartan treatment (eg, 20 mg / kg / day) significantly reduced collagen levels in treated tumors. FIG. 5B is a bar graph summarizing the effects after 2 weeks of treatment with 20 mg / kg / day of losartan; losartan treatment was performed with collagen I immunostaining in L3.6pl (p <0.03) and FVB MMTV PyVT, Reducing significantly by 50% (p <0.05) and 47% (p <0.05), respectively. FIG. 5C is a photographic panel showing collagen I and nuclear immunostaining in tumor sections in HSTS26T and Mu89 tumors treated with control and losartan (20 mg / kg / day). Note that 200 μm from the edge of the HSTS26T tumor has no detectable decrease in collagen I immunostaining. This phenomenon is less pronounced in treated Mu89 tumors where there was persistent staining in both the edge and central tumor areas. Scale bar = 100 μm. FIG. 5D is a bar graph summarizing the effects of losartan that significantly reduced collagen I immunostaining in HSTS26T and Mu89 by 44% (p <0.02) and 20% (p <0.05), respectively. Panel of bar graphs showing the effect of losartan on TSP-1, active TGFβ1, total TGFβ1, and collagen I in HSTS26T tumors. Treated animals received drinking water containing losartan (15 mg / kg / day). Tumors were excised, homogenized after 2 weeks of treatment, and analyzed by ELISA for total and active TGFβ1 levels. Note a 3.5-fold decrease in TSP-1 after losartan treatment, a 4-fold decrease in active TGFβ1, and a 2-fold decrease in collagen 1 (p <0.05). FIG. 7A shows the effect of losartan with reduced tumor TSP-1 immunostaining in both MU89 and HSTS26T tumors. In HSTS26T tumors, changes in TSP-1 after losartan treatment correspond to changes in collagen I immunostaining; TSP-1 levels decrease at the tumor center but remain high within 200 μm from the tumor edge. The TSP-1 margin was larger in the MU89 tumor (500 μm from the edge). Scale bar = 100 μm. FIG. 7B is a bar graph summarizing the effects of losartan treatment that significantly reduced TSP-1 immunostaining in HSTS26T and MU89 tumors by 73% (p <0.04) and 24% (p <0.03), respectively. Figure 2 shows the effect of losartan with increased delivery of nanoparticles and nanotherapeutics. FIG. 8A shows two photographs (control and losartan) and a bar graph summarizing the dispersion of intratumoral (i.t.) injected 100 nm diameter nanoparticles in HSTS26T tumors. Losartan significantly increased the dispersion of i.t. injected nanoparticles in both tumor types (p <0.001) (1.5 fold in HSTS26T and 4 fold in Mu89). Analysis of the dispersion pattern shows that there were fewer intratumoral nanoparticles in the control tumor, and a majority of the nanoparticles were back from the needle track and accumulated on the tumor surface. In contrast, treated tumors have a significant number of intratumoral nanoparticles. Scale bar = 100 μm. FIG. 8B shows two photographs (control and losartan) and a bar graph summarizing the dispersion of viral infection 24 hours after intratumoral injection of HSV expressing green fluorescent protein. Although HSV infection in control tumors is limited to cells in close proximity to the injection site, tumors treated with losartan have a wider spread of HSV infection within the tumor. Scale bar = 1mm. Losartan significantly increased viral spread in HSTS26T and Mu89 tumors (p <0.05). FIG. 8C shows two photographs (control and losartan) and a bar graph summarizing the dispersion of 100 nm diameter nanoparticles injected intravenously (i.v.) in L3.6pl tumors. The nanoparticles are localized around the perfused blood vessel. In tumors treated with losartan, there is a 2-fold increase in nanoparticle content (p <0.05) compared to control tumors. Scale bar = 100 μm. FIG. 6 is a bar graph showing the change in diffusion coefficient in HSTS26T tumor after losartan treatment. IgG diffusion coefficient was measured in HSTS26T tumors implanted in dorsal window chambers of SCID mice. Treated animals received losartan by i.p. injection (40 mg / kg / day) and control animals received saline. The results show a significant increase (p <0.04) in diffusion coefficient as measured by multiphoton fluorescence post-fading recovery (FRAP). FIG. 2 is a representative dispersion profile showing the fraction of injected nanospheres present as a function of distance from tumor blood vessels (depth of penetration). Nanosphere penetration depth was analyzed in frozen sections from tumors excised 24 hours after intravenous nanosphere injection. The mean characteristic penetration length increased from 18 ± 5 μm (mean ± SE) in controls to 37 ± 6 μm in tumors treated with losartan. In 6 control tumors and 6 treated tumors, 10 areas were analyzed for each tumor. FIG. 6 shows the effect of losartan that significantly delayed the growth of tumors treated with DOXIL® or HSV. FIGS. 11A-11B are the resulting polylines from mice bearing HSTS26T tumors (A) and Mu89 tumors (B) treated for 2 weeks with either losartan or saline prior to it injection of HSV A graph is shown. Losartan alone did not affect the growth of Mu89 or HSTS26T tumors. The augmentation delay was significantly longer in HSTS26T tumors treated with losartan and HSV compared to tumors treated with single HSV. HSV i.t. injection did not delay the growth of Mu89 tumors, but treatment with the combination of losartan and HSV significantly delayed the growth of Mu89 tumors. FIG. 11C shows that mice that received losartan treatment prior to ivDOXIL® infusion (Losartan and DOXIL®) received DOXIL® alone (DOXIL® alone) Figure 9 shows the effect on tumor volume in L3.6pl tumors that had small tumors. Note that there is no difference in tumor size between mice treated with saline and mice treated with losartan. FIG. 11D is an image showing a clear size difference between a control tumor (left column) and a tumor treated with losartan (right column) one week after DOXIL® injection. Scale bar = 1cm. One week after DOXIL® injection, tumors treated with losartan (right column) were smaller than control tumors (left column). Figure 2 shows the relationship between collagen structure and viral infection and necrosis. FIG. 12A, in the Mu89 tumor, collagen bundles are seen around the tumor periphery. Sometimes these bundles enter the tumor (black arrows) and divide the tumor into separate compartments. These compartments appear to limit HSV migration; it is evident from the necrotic area contained within the area bounded by the collagen bundle. When these tumors were treated with losartan, the collagen bundles at the periphery of the tumor remained intact, but the processes became more disordered (inset). This probably allowed virus propagation and necrosis to extend beyond the boundaries. Scale bar = 100 μm. In FIG. 12B, HSTS26T, a dense reticulated collagen network restricted viral infection to the area immediately surrounding the injection point. Losartan treatment caused a decrease in the density of the network, possibly allowing virus particles to infect larger areas and thus more tumor cells. Arrows indicate live cells and virus-infected cells, respectively. Scale bar = 10 μm. Schematic representation of virus distribution and infection in Mu89 tumor (A) and HSTS26T tumor (B). The schematic diagram shows how different collagen network structures affect virus propagation and distribution. Collagen fibers (1) restrict migration of viral particles (round spheres, 2) and uninfected (4) infection of cancer cells (3). FIG. 13A, in the Mu89 tumor, the collagen bundle divides the tumor into isolated areas that cannot be surpassed by the viral particles. Losartan treatment destabilizes the collagen bundle and allows virus particles to move from one region to another. In FIG. 13B, HSTS26T tumor, the collagen structure is a mesh screen. Viral particles can propagate through the sieve but do not expand very far from the injection site. Losartan treatment significantly destabilizes the network structure in the internal region of the tumor, allowing the virus to propagate and infect larger areas. FIG. 14A shows viral infection (HSV immunostaining) and necrosis 21 days after HSV injection in HSTS26T and MU89. Complete tumor area, necrotic hematoxylin staining, and HSV immunostaining are shown. Necrotic areas are indicated by black arrows. There was no detectable difference in necrotic area between HSTS26T and MU89, but in MU89 the necrosis was restricted to a specific area, and in HSTS26T tumors the entire necrotic tissue (bound by HSV immunostaining). Is present). Scale bar = 2mm. FIG. 14B is a bar graph showing that there is a 2-fold increase in necrosis (p <0.05) in tumors that received losartan (both HSTS26T and MU89) prior to HSV injection. In vivo growth rate for HSTS26T and MU89 after losartan treatment. Tumors were excised and stained for Ki67 to assess growth rate. There was no statistically significant difference in positive Ki67 staining after losartan treatment in HSTS26T and MU89 tumors. However, there was a significant difference in proliferation between the two tumor types and the number of Ki67 positive cells was three times higher in HSTS26T tumors. The results of PCR analysis of AGTR1 expression in CAF cells, MU89 cells, and HSTS26T cells are shown. MU89 cells and CAF express AGTR1, but not HSTS26T cells. HUVEC was used as a positive control. GAPDH levels revealed that all three samples had approximately the same amount of cDNA. FIG. 5 shows the effect of normalizing the tumor microenvironment of angiotensin inhibition by AT ₁ blockers or ACE inhibitors. Studies using ARB and losartan are shown. Angiotensin inhibition reduces (A) stromal matrix density in breast tumors (MMTV) and pancreatic tumors (L3.6PL) in mice and (B) compressive stress in breast tumors (E0771) and pancreatic tumors (Pan-02) Reduce. (C) This increases the fraction of perfused blood vessels (arrows) in the tumor (E0771 is shown), (D) Normalized blood vessels, more efficient and effective in drug and oxygen delivery Bring the network (E0771 is shown). FIG. 5 shows the effect of angiotensin inhibition by AT ₁ blockers or ACE inhibitors to improve drug transport and dispersion in tumors. A study using ARB, losartan is presented here. Through tumor normalization, angiotensin inhibition (A) improves tumor oxygenation through enhanced perfusion (E0771 is shown) and (B) allows drug delivery to blood vessels more rapidly. Reorganization of the stromal matrix (C) also improves nanoparticle penetration in fibrogenic tumors (denoted L3.6PL) (D). FIG. 5 shows the effect of angiotensin inhibition by AT ₁ blockers or ACE inhibitors to improve the effectiveness of cancer treatment. Studies using ARB and losartan are shown. Angiotensin inhibition given in combination with chemotherapy (A) improves the efficacy of the low MW chemotherapeutic drug doxorubicin in breast cancer models, (B) slows tumor growth, and (C) increases animal survival ( E0771 is shown). Similarly, angiotensin inhibition (D, E) can cause nanoparticle DOXIL in pancreatic tumors (denoted L3.6PL), for example, by reducing tumor weight (D) and / or tumor size or volume (E) Improve the effectiveness of (registered trademark).

Detailed Description of the Invention The present invention is based at least in part on the discovery that losartan, an antihypertensive agent, improves the delivery and efficacy of cancer therapeutics.

  An abnormal matrix of tumors limits the delivery of nanotherapeutic agents in many types of cancer, such as pancreatic cancer, breast cancer, lung cancer, colorectal cancer. Fibrous tissue overgrowth prevents nanotherapeutic drug migration in tumors by two mechanisms: viscoelastic and steric hindrance. Fibrous tissue is highly viscoelastic, ie very dense and stiff, thus slowing the movement of these drugs to a fraction of typical speed. This tissue is basically a very dense network with small pores approximately the same size as the nanotherapeutics, thus not giving much space for these drugs and blood vessels (in the case of intravenous injection) Or they are often stopped by restricting them near the injection site (in the case of intratumoral injection). This barrier is found in all solid tumors, except perhaps brain tumors, but is most prominent in pancreatic cancer, breast cancer, lung cancer, and colorectal cancer. Nanotherapeutics are particularly disturbed by fibrous tissue because of their large size compared to the pores that form the tumor microenvironment.

In certain embodiments, Applicants have shown that losartan prevents the production of matrix molecules such as collagen that are components of dense networks of fibrous tissue. Losartan is thought to act on fibroblasts and tumor cells by inhibiting the TGFβ and CTGF pathways and thus limiting their profibrotic activity. It occurs by inhibiting the activity of angiotensin II type 1 receptor (AT ₁ ) that is highly expressed in both fibroblasts and tumor cells in a variety of cancers. Thus, losartan inhibits downstream activities of AT ₁ in various signaling pathways, including activation of TGFβ and CTGF. Since these two pathways promote the production of collagen and other components of fibrous tissue, their inhibition will allow fibrosis to subside. As a result, the tissue becomes much more like normal surrounding organs and is therefore easier to penetrate.

  It is shown herein that treatment with losartan significantly reduces collagen levels, a marker of fibrosis, in several types of tumors, including pancreatic, breast, skin, and soft tissue tumors. In addition, the reduction in fibrosis results in improved mobility of nanotherapeutic drugs in the tumor, allowing them to penetrate the tumor more easily and allow these drugs to be more widely dispersed throughout the tumor And become more effective in resisting tumor growth. Therefore, losartan makes nanotherapeutics more effective against cancer.

  More specifically, Applicants have noted that losartan normalizes collagen, which is the stromal matrix of several solid tumors, thereby penetrating chemotherapeutic drugs such as high molecular weight (eg, nano) chemotherapeutic drugs Found to facilitate. For example, losartan reduces collagen I levels in cancer-associated fibroblasts (CAF) isolated from breast cancer biopsies and is a substrate in human breast, pancreas, and skin tumor fibrosis models in mice Caused a dose-dependent decrease in collagen. Losartan also improved the dispersion, therapeutic efficacy, and / or penetration of nanoparticles (eg, oncolytic herpes simplex virus (HSV) and PEGylated liposomal doxorubicin (DOXIL®)).

  Low molecular weight therapeutics that are much smaller than nanotherapeutics are not limited by the interstitial matrix barrier, but are similarly affected by other barriers such as abnormal and collapsed blood vessels.

  In other embodiments, Applicants have determined that losartan facilitates vascular decompression and thus improves tumor perfusion and delivery of low molecular weight chemotherapeutic agents through vascular normalization, and thus delivery of radiation and chemotherapeutic agents. I found it easy.

  Thus, these agents improve delivery of small molecules as small as oxygen, a radiochemical sensitizer, through vascular normalization (FIGS. 18A-18B) and enhance penetration of larger agents through stromal matrix normalization. (FIGS. 18C and 18D). Through this repair of the entire tumor microenvironment, these agents enhance the effectiveness of low molecular weight chemotherapeutic and nanotherapeutic drugs in breast and pancreatic cancer models, resulting in reduced tumor growth and longer animal survival (Figure 19A-19E). Thus, angiotensin inhibitors (eg, angiotensin receptor blockers) and ACE inhibitors can enhance the delivery of therapeutic agents, and thus low molecular weight chemotherapeutic agents, small molecule chemotherapeutic agents, biologics, nucleic acid agents And has broad applicability for combination therapy with all classes of anticancer agents, including nanoparticle therapy.

  Angiotensin blockers show a number of advantages over other approaches. Anti-angiogenic treatment normalizes only the vasculature and is approved only for a limited number of indications. On the other hand, ARB and ACE-I are approved by the FDA as antihypertensive agents with manageable adverse effects. Matrix degrading enzymes that can normalize the collagen matrix are not selective for tumors and may increase invasion and metastasis. ARB and ACE-I do not have significant complications associated with matrix remodeling in normal tissues and are safe as antihypertensive drugs. The small molecule drugs ARB and ACE-I are further mediated through nanovectors containing chemotherapeutic drugs (eg, liposomes, nanoparticles) to further limit toxicity and enhance tumor localization May be delivered. Anti-angiogenic drugs, the only FDA-approved adjuvant that enhances drug delivery to tumors, can reduce the size of “pores” in the vessel wall, thus improving delivery for larger particles It is not possible. Angiotensin blockers can improve delivery for all classes of anti-tumor diagnostics and anti-tumor treatments.

  Accordingly, methods and compositions that improve the delivery and / or efficacy of cancer therapeutics are disclosed. By administering an antihypertensive agent to a subject as a single agent or in combination with a cancer therapeutic agent (eg, a therapeutic agent ranging in size from a large nano-therapeutic agent to a low molecular weight chemotherapeutic agent and / or oxygen) Disclosed are methods and compositions for treating or preventing cancer (eg, solid tumors such as fibrogenic tumors).

  Certain terms are defined first.

  “About” and “approximately” shall generally mean an acceptable degree of error for the measured quantity, taking into account the nature or accuracy of the measurement. Exemplary degrees of error are within 20 percent (%) of a given value or range of values, typically within 10%, more typically within 5%, within 4%, within 3%, Within 2% or within 1%.

  “Delivery”, as used herein with respect to the delivery of an agent to a tumor, has the desired effect and therefore has a desired effect on the tumor vasculature, tumor stroma matrix, or tumor cells or tumor-related cells (eg, fibers The agent is placed in close proximity to one or more (or all) of the blast cells. The agent can be, for example, a cancer treatment (eg, a cancer treatment described herein), a diagnostic agent, or an imaging agent. Unless otherwise indicated, the term “agent” can include one, two, or more agents, as used generically herein.

  In one embodiment, the cancer therapeutic agent includes, for example, one or more of small molecule drugs, protein drugs, nucleic acid drugs, oncolytic viruses, vaccines, antibodies or fragments thereof, or combinations thereof. The cancer therapeutic agent may be “free” or packaged or formulated into a delivery vehicle, eg, a particle, eg, a nanoparticle (eg, lipid nanoparticle, polymer nanoparticle, or virus particle). It may be. The delivery of the therapeutic agent is characterized by the therapeutic agent being placed in close proximity to the cell in order to modify the activity of the cell, for example to kill the cell and / or reduce its ability to divide.

  In other embodiments, the agent is a diagnostic or imaging agent (eg, one or more of a radiation agent, NMRA agent, contrast agent, etc.). The diagnostic or imaging agent may be “free” or packaged or formulated in a delivery vehicle. Delivery of a diagnostic or imaging agent is characterized by the agent being placed in close proximity to the target cell or tissue to allow detection of the target cell or tissue.

In embodiments, an increase (improvement) in delivery (compared to delivery that is identical or similar except that it is performed in the absence of AHCM) includes:
Delivery of the agent to the tumor vasculature, or an increase in the amount or concentration in the tumor vasculature;
Delivery of an agent to a tumor, eg, tumor vasculature stromal matrix, or an increase in amount or concentration in a tumor, eg, tumor vasculature stromal matrix;
Delivery of the agent to tumor cells or tumor-related cells (eg, fibroblasts), or an increase in amount or concentration in tumor cells or tumor-related cells (eg, fibroblasts);
Increased flow rate in the tumor vasculature, eg, agent flow rate;
Improvement (or normalization) of vascular morphology (eg, becoming less tumor-like);
Decompression of tumor vasculature;
An increase in the pore size or diffusion rate of the agent in the tumor, eg, the stromal matrix;
Increased drug perfusion in tumors, eg, stromal matrix;
Wider and / or more homogeneous distribution of the agent throughout the tumor;
A broader and / or more homogeneous distribution of the agent throughout the tumor stromal matrix;
An increase in the proportion of the agent in the tumor, eg tumor stromal matrix, relative to non-tumor tissue, eg peripheral blood;
Inhibition of the TGFβ pathway in tumors, eg, tumor vasculature stromal matrix;
Inhibition of the CTGF pathway in tumors, eg, tumor vasculature stromal matrix;
Inhibition of angiotensin II type 1 receptor activity;
One or more of a reduction in fibrosis in a tumor, eg, tumor vasculature stromal matrix; or a reduction in collagen or collagen deposition in a tumor, eg, a matrix of tumor vasculature stroma.

  In some embodiments, increased delivery (ie, improvement) (as compared to delivery that is identical or similar except that performed in the absence of AHCM) is dispersed in at least a portion of the tumor Increasing the amount of agent to be included can also be included. In some embodiments, the increase in the amount of agent delivered to the tumor in the presence of AHCM may be dispersed homogeneously or heterogeneously throughout the tumor.

  “Efficacy” as used herein with respect to treatment, eg, cancer treatment, may be detectable or undetectable, alleviating symptoms, reducing the degree of disease, Characterized as the degree to which a treatment has a desired effect, including, but not limited to, a stable state of disease, delay or slowing of disease progression, amelioration or alleviation of disease state, and remission (partial or complete) .

  With respect to the efficacy of cancer therapy, the improvement in efficacy is an increase in the anti-tumor effect of the cancer therapy compared to a treatment that is the same or similar except that it is performed in the absence of treatment with AHCM, and / Or it may be characterized by one or more of the reduction of unwanted side effects (eg, toxicity) of cancer treatment. In one embodiment, an increase in the anti-tumor effect of cancer treatment includes inhibition of the growth of primary or metastatic tumors; a decrease in the amount or volume of primary or metastatic tumors; the size or number of metastatic lesions Reduction; inhibition of the development of new metastatic lesions; reduction of one or more of the volume or metabolism of a non-invasive tumor; provision of prolonged survival; provision of prolonged progression-free survival; prolonged progression One or more of providing downtime; and / or enhancing quality of life is included.

  In some embodiments, with respect to cancer therapy combined with AHCM, the term “improved efficacy” as used herein is the primary in cancer therapy alone (ie, in the absence of AHCM). At least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least, compared to a decrease in the growth of a sex or metastatic tumor About 70%, at least about 80%, at least about 90%, at least about 95%, up to 100%, an increase in the decrease in primary or metastatic tumor growth can be referred to. In some embodiments, administration of AHCM in combination with a cancer treatment is at least compared to a decrease in the growth of primary or metastatic tumors in the cancer treatment alone (ie, in the absence of AHCM). About 1 fold, at least about 2 fold, at least about 3 fold, at least about 5 fold, at least about 6 fold, at least about 7 fold, or more, can increase the decrease in the growth of primary or metastatic tumors . Methods for monitoring tumor growth in vivo are well known in the art, including but not limited to x-ray, CT scan, MRI, and other art-recognized medical imaging methods. .

  In some embodiments, with respect to cancer therapy in combination with AHCM, the term “improved efficacy” as used herein refers to perfusion of an anticancer agent alone (ie, in the absence of AHCM). For example, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, At least about 90%, at least about 95%, and up to 100% of an increase in perfusion to the tumor of an anti-cancer agent (eg, a low molecular weight therapeutic agent or a nanotherapeutic agent such as DOXIL®). In some embodiments, administration of AHCM in combination with cancer treatment is at least about 1 fold, at least about 2 fold, at least compared to the perfusion efficiency of the anticancer agent alone (ie, in the absence of AHCM). About 3 times, at least about 5 times, at least about 6 times, at least about 7 times, or more, perfusion of tumors with anticancer agents (eg, low molecular weight therapeutic agents or nanotherapeutic agents such as DOXIL®) Can be increased. Methods for monitoring tumor perfusion in vivo, including but not limited to positron emission tomography (PET) and ultrasound or contrast ultrasound, are well established in the art.

  In some embodiments, with respect to cancer treatment in combination with AHCM, the term “improved efficacy” as used herein is that the cancer treatment is administered alone (ie, in the absence of AHCM). For example, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, compared to a decrease in the expression level of at least one cancer biomarker At least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, up to 100% (eg, as a blood sample, serum sample, plasma sample, or tissue biopsy Increase in the expression level of at least one cancer biomarker (in a biological sample). In some embodiments, administration of AHCM in combination with a cancer treatment reduces the expression level of at least one cancer biomarker when the cancer treatment is administered alone (ie, in the absence of AHCM). In comparison, at least about 1 fold, at least about 2 fold, at least about 3 fold, at least about 5 fold, at least about 6 fold, at least about 7 fold, or more (e.g., blood sample, serum sample, plasma sample, Alternatively, the reduction in the expression level of at least one cancer biomarker (in a biological sample such as a tissue biopsy) can be increased. Examples of serum / plasma cancer biomarkers can include, but are not limited to, TGFβ1, TGFβ2, CTGF, TSP-1, Collagen I, Collagen II, Collagen III, Collagen IV. The expression level of serum / plasma cancer biomarkers can be determined at the transcript and / or protein level using art-recognized analytical methods such as PCR, Western blot, ELISA, and / or immunostaining. Can be measured.

  “Blood pressure” is generally classified based on systolic blood pressure and diastolic blood pressure. “Systolic blood pressure” or Psys refers to the blood pressure in the blood vessels during heartbeat. “Diastolic blood pressure” or Pdias refers to the pressure during the heartbeat. Measurements of systolic blood pressure or diastolic blood pressure that are higher than the normal values allowed for the individual's age are classified as prehypertension or hypertension. Measurements of systolic blood pressure or diastolic blood pressure that are lower than the normal value allowed for the age of the individual are classified as hypotension. “Normal” systolic pressure for adults is typically in the range of 90-120 mmHg; “normal” diastolic pressure is generally in the range of 60-80 mmHg. In the population, mean blood pressure (Psys / Pdias ratio) can range from 110/65 to 140/90 mmHg for adults; 95/65 mmHg for 1 year olds and 100/65 mmHg for 6-9 years old.

  Hypertension is pre-hypertension (120/80 to 139 / 89mmHg); Grade I hypertension (140/90 to 159 / 99mmHg), Grade II hypertension (over 160 / 100mmHg), Systolic hypertension (140 / 90mmHg) Have several subclasses including Systolic hypertension refers to those in which systolic pressure is elevated but diastolic pressure is normal, and is common among elderly people. These classifications are made after averaging the patient's resting blood pressure readings obtained from more than one visit.

  Hypertension is generally diagnosed based on persistently high blood pressure. In general, this requires three separate sphygmomanometer measurements at least one week apart. Often this requires three separate visits to a medical institution. The initial assessment of hypertensive patients should include a complete medical history and physical examination.

  As used herein, “hypertension” or “hypertension” refers to prehypertension or hypertension having a systolic pressure of 120 or higher (typically 140 or higher) and a diastolic pressure of 80 or higher. Refers to the stage of the disease (where all blood pressures are expressed as mmHg).

  As used herein, the term “mean arterial pressure” (MAP) is recognized in the art and refers to the average in the cardiac cycle, cardiac output (CO), systemic vascular resistance. (SVR), and central venous pressure (CVP) (MAP = (CO × SVR) + CVP). When a normal resting heart rate is present, MAP can be approximately determined from measurements of systolic pressure (Psys) and diastolic pressure (Pdias), where MAP is approximately Pdias + 1/3 (Psys-Pdias).

A “hypertensive agent”, as used herein, is typically a patient (eg, hypertension) when administered at a selected dose (referred to herein as a “hypertensive dose”). Patient) refers to an agent (eg, compound, protein) that lowers blood pressure. Antihypertensive agents are routinely used clinically to treat patients with hypertension at doses known in the art. Exemplary antihypertensive agents include, but are not limited to, renin angiotensin aldosterone antagonists (“RAAS antagonists”), angiotensin converting enzyme (ACE) inhibitors, and angiotensin II receptor blockers (AT ₁ blockers). Not. Some exemplary antihypertensive doses of these agents are also disclosed herein.

A “non-hypertensive dose” as used herein refers to a dose of an antihypertensive agent that is typically less than the lowest dose that would be used to treat a patient due to hypertension. In one embodiment, the non-hypertensive dose has one or more of the following characteristics:
Does not substantially reduce the blood pressure of a subject, eg, a hypertensive subject, eg, mean arterial blood pressure;
The subject's mean arterial blood pressure is <1%, <5%, <10%, <15%, <20%, <25%, <30%, <35%, <40%, <45%, <50% Reduce;
Less than 1%, less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 35%, less than 40% of the reduction caused by the standard of medical antihypertensive dose for that AHCM Reduce blood pressure to less than 45%, less than 50%, less than 60%, less than 70%, less than 80%, less than 90%, or less;
The subject's blood pressure will be in the normal range, for example, its AHCM dose that will be in 120 systolic and 80 diastolic, or the subject's blood pressure will be in the range of 120 +/- 5 systolic and 80 +/- 5 diastolic More than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% of the dose will be Less; or less than the standard of medical hypotensive dose.

  In certain embodiments, the ability of a dose to meet one or more of these criteria is determined after a preselected number of times, e.g., after 1, 2, 5, or 10 doses. Or it can be measured after dosing sufficient to achieve steady state levels, eg plasma levels.

“AHCM”, as used herein, can be an agent having one or more of the following properties:
Is an antagonist of the renin angiotensin aldosterone system (“RAAS antagonist”),
An angiotensin converting enzyme (ACE) inhibitor,
Angiotensin II receptor blocker (AT ₁ blocker),
A thrombospondin 1 (TSP-1) inhibitor,
It is a transforming growth factor β1 (TGFβ1) inhibitor or a connective tissue growth factor (CTGF) inhibitor.

“Treatment” of a tumor, as used herein, typically
Inhibition of the growth of primary or metastatic tumors;
Reduction in the volume or volume of primary or metastatic tumors;
Reduction in the size or number of metastatic lesions;
Inhibition of the development of new metastatic lesions;
Reduction of one or more of the volume or metabolism of a non-invasive tumor;
Providing extended survival;
Providing extended progression-free survival;
One or more of providing extended progression stoppage time; and / or enhancing quality of life.

  Various aspects of the invention are described in further detail below. Additional definitions are given throughout this specification.

Antihypertensive and / or collagen modifier (AHCM agent)
In certain embodiments, an AHCM agent used in the methods and compositions of the invention comprises an antagonist of the renin angiotensin aldosterone system (“RAAS antagonist”), an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker ( Selected from one or more of AT ₁ blocker, thrombospondin 1 (TSP-1) inhibitor, transforming growth factor β1 (TGFβ1) inhibitor, and connective tissue growth factor (CTGF) inhibitor obtain. The method may comprise one, two, three, or more AHCM agents alone or in combination with one or more cancer therapeutics.

  Exemplary antagonists of the renin angiotensin aldosterone system (RAAS) include aliskiren (TEKTURNA®, RASILEZ®), remixylene (Ro 42-5892), enalkylene (A-64662), SPP635, and their Derivatives are included, but are not limited to these.

  Exemplary angiotensin converting enzyme (ACE) inhibitors include benazepril (LOTENSIN®), captopril (CAPOTEN®), enalapril (VASOTEC®), fosinopril (MONOPRIL®), Lisinopril (PRINIVIL (registered trademark), ZESTRIL (registered trademark)), Moexipril (UNIVASC (registered trademark)), Perindopril (ACEON (registered trademark)), Quinapril (ACCUPRIL (registered trademark)), Ramipril (ALTACE (registered trademark)) , Trandolapril (MAVIK®), and derivatives thereof.

Exemplary angiotensin II receptor blockers (AT ₁ blockers) include losartan (COZAAR®), candesartan (ATACAND®), eprosartan mesylate (TEVETEN®), EXP 3174 , Irbesartan (AVAPRO®), L158,809, olmesartan (BENICAR®), salaracin, telmisartan (MICARDIS®), valsartan (DIOVAN®), and derivatives thereof However, it is not limited to these.

In one embodiment, the AT ₁ blocker is losartan or a derivative thereof. Losartan is an antihypertensive with minimal safety risk (Johnston CI (1995) Lancet 346: 1403-1407). In addition to antihypertensive properties, losartan is also an antifibrotic agent that has been shown to reduce the incidence of heart and kidney fibrosis (Habashi JP, et al. (2006) Science 312: 117- 121; and Cohn RD, et al. (2007) Nat Med 13: 204-210). The antifibrotic effect of losartan is an active transforming growth factor β1 (down-regulation mediated by angiotensin II type I receptor (AGTR1) of TGFβ1 activators such as thrombospondin 1 (TSP-1) ( Inhibition of TGFβ1) levels is caused in part (Habashi JP, et al. (2006) Science 312: 117-121; Cohn RD, et al. (2007) Nat Med 13: 204-210; Lavoie P, et al (2005) J Hypertens 23: 1895-1903; Chamberlain JS (2007) Nat Med 13: 125-126; and Dietz HC (2010) J Clin Invest 120: 403-407).

  Exemplary thrombospondin 1 (TSP-1) inhibitors include, but are not limited to, ABT-510, CVX-045, LSKL, and derivatives thereof.

  Exemplary transforming growth factor β1 (TGFβ1) inhibitors include CAT-192, Fresolimumab (GC1008), LY 2157299, Peptide 144 (P144), SB-431542, SD-208, US Pat. No. 7,846,908 and US Pat. Examples include, but are not limited to, the compounds described in Application Publication No. 2011/0008364, as well as their derivatives.

  Exemplary connective tissue growth factor (CTGF) inhibitors include the compounds described in DN-9693, FG-3019, and European Patent Application Publication No. 1839655, US Pat. No. 7,622,454, and derivatives thereof. However, it is not limited to these.

  Exemplary beta blockers include atenolol (TENORMIN®), betaxolol (KERLONE®), bisoprolol (ZEBETA®), metoprolol (LOPRESSOR®), metoprolol long-term release ( TOPROL XI (registered trademark), nadolol (CORGARD (registered trademark)), propranolol (INDERAL (registered trademark)), long-acting propranolol (INDERAL LA (registered trademark)), timolol (BLOCADREN (registered trademark)), acebutolol (SECTRAL (R)), penbutolol (LEVATOL (R)), pindolol, carvedilol (COREG (R)), labetalol (NORMODYNE (R), TRANDATE (R)), and derivatives thereof However, it is not limited to these.

  In one embodiment, the AHCM agent is a TGFβ1 inhibitor, such as an anti-TGFβ1 antibody, a TGFβ1 peptide inhibitor. In certain embodiments, the TGFβ1 inhibitor is CAT-192, Fresolimumab (GC1008), LY 2157299, Peptide 144 (P144), SB-431542, SD-208, US Patent No. 7,846,908 and US Patent Application Publication No. 2011 / [0008] It is selected from one or more of the compounds described in No. 364, or their derivatives.

  Appropriate doses for administration of AHCM agents can be assessed based on medical antihypertensive dose standards for AHCM agents available in the art.

Exemplary standards for medical doses and medical dosage formulations for antihypertensive and anti-heart failure for AT ₁ inhibitors in humans are as follows: about 25-100 mg / day of losartan (about 12.5 mg, 25 mg, 50 mg Or available in dosage forms for oral administration containing 100 mg losartan; containing 4 to 32 mg / day candesartan (ATACAND®) (eg, containing 4 mg, 8 mg, 16 mg, or 32 mg candesartan) Oral dosage forms containing 400-800 mg / day eprosartan mesylate (TEVETEN®) (eg 400 mg or 600 mg eprosartan) 150-300 mg / day irbesartan (AVAPRO®) (eg available in dosage forms for oral administration containing 150 mg or 300 mg irbesartan); 20-40 mg / day Olmesartan (BENICAR (Register )) (Available in oral dosage forms containing 5 mg, 20 mg, or 40 mg olmesartan); 20-80 mg / day telmisartan (MICARDIS®) (eg, 20 mg, 40 mg, or Available in dosage forms for oral administration containing 80 mg telmisartan); and 80-320 mg / day valsartan (DIOVAN®) (eg, containing 40 mg, 80 mg, 160 mg, or 320 mg valsartan) Available in dosage forms for oral administration).

  Exemplary standards for medical doses and medical dosage formulations for antihypertensive and anti-heart failure for ACE inhibitors in humans are as follows: 10-40 mg / day benazepril (LOTENSIN®) (Lotensin ( Benazepril) is supplied as tablets for oral administration containing 5 mg, 10 mg, 20 mg or 40 mg benazepril hydrochloride); 25-100 mg / day captopril (CAPOTEN®) (approximately 12.5 mg Available in oral dosage forms containing 25 mg, 50 mg, or 100 mg captopril); 5-40 mg / day enalapril (VASOTEC®) (2.5 mg, 5 mg, 10 mg, or 20 mg Available in oral dosage form containing enalapril); 10-40 mg / day fosinopril (MONOPRIL®) (10 mg, 20 mg, or 40 mg fosinopril containing oral fosinopril) Available in the form); 10-40 mg / Of lisinopril (PRINIVIL®, ZESTRIL®) (available in dosage forms for oral administration containing 2.5 mg, 5 mg, 10 mg, 20 mg, 30 mg, or 40 mg of lisinopril); 7.5-30 mg / Day moexipril (UNIVASC®) (available in dosage forms for oral administration containing 7.5 mg or 15 mg moexipril); 4-8 mg / day perindopril (ACEON®) (eg Available in oral dosage forms containing 2 mg, 4 mg, or 8 mg perindopril), 10-80 mg / day quinapril (ACCUPRIL®) (5 mg, 10 mg, 20 mg, or 40 mg quinapril Available in dosage forms for oral administration containing); 2.5-20 mg / day ramipril (ALTACE®) (oral administration containing 1.25 mg, 2.5 mg, 5 mg, or 10 mg ramipril) Available in dosage form); 1-4 mg / day trandolapril (MAVIK®) (available in dosage forms for oral administration containing 1 mg, 2 mg, or 4 mg of trandolapril).

  In one embodiment, the AHCM agent is administered in standard medical doses and standard medical dosage formulations for antihypertensive and anti-heart failure, such as the doses or dosage formulations described herein.

  In certain embodiments, a non-hypertensive dose or non-hypertensive dosage formulation of an AHCM agent, such as a standard medical dose or a dose of AHCM agent less than a standard medical dosage formulation, is desirable. In one embodiment, the non-hypertensive dose or non-hypertensive dosage formulation has a minimal effect on blood pressure in hypertensive subjects (eg, less than 20%, less than 10%, mean arterial blood pressure in hypertensive subjects, 5 %, Or less). In certain embodiments, the AHCM agent is dosed at less than 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 of a standard for a medical antihypertensive dose (eg, a lower standard for medical dose) Or administered in a dosage formulation. In one embodiment, the dose or dosage formulation is, for example, 0.01-0.9 times, 0.02-0.8 times, 0.05-0.7 times, 0.1-0.1 times the standard medical dose or standard medical dosage formulation used for antihypertensive or anti-heart failure. The range is 0.5 times and 0.1 to 0.2 times. Standard medical doses or standard medical dosage formulations of AHCM are available in the art, some of which are exemplified herein.

  In still other embodiments, the AHCM agent is a standard medical dose used for antihypertensive or anti-heart failure or a dosage or formulation greater than a standard medical dosage formulation (eg, standard medical dose used for anti-hypertensive or anti-heart failure) Doses or dosage forms that are 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times or more. In one embodiment, the dose or dosage formulation is, for example, 1.1-10 times, 1.5-5 times, 1.7-4 times, or 2 times the standard medical dose or standard medical dosage formulation used for antihypertensive or anti-heart failure. ~ 3 times the range. Standard medical doses or standard medical dosage formulations of AHCM are available in the art, some of which are exemplified herein.

  Standard medical doses and standard medical dosage forms for a number of AHCMs, eg, losartan, are provided herein. In one aspect, the dose and / or dosage form is lower (or higher) than the standard medical dose and / or standard medical dosage form. In exemplary embodiments, it is 0.01 times, 0.02 times, 0.05 times, 0.1 times, 0.2 times, 0.5 times, 0.7 times, 0.8 times, 0.9 times lower than the standard medical dose or standard medical dosage form. In an embodiment, the dose or dosage form contains an amount of AHCM that is within the standard medical dose and / or the reduced amount of the standard medical dosage form. For example, an AHCM dosage form that is 0.01-0.9 times, 0.02-0.8 times, 0.05-0.7 times, 0.1-0.5 times, 0.1-0.2 times the standard medical dosage or standard medical dosage form. In certain embodiments, as long as the value of the dose or dosage form is lower than the standard medical dose or standard medical dosage form, the range of the dose or dosage form is 0.5 to 0.5 of the reduced dose or dosage form described herein. 2.0 times. For example, the standard medical dosage form for losartan is 12.5 mg. Thus, in embodiments, the dosage form comprises 0.125 mg (.01 × 12.5 mg); 0.625 mg (0.05 × 12.5 mg); 1.25 mg (0.1 × 12.5 mg); 2.5 mg (0.2 × 12.5 mg); or 6.25 mg ( 0.5 × 12.5 mg). In one embodiment, the AHCM dosage form is 0.5-2.0 (.125 mg) = 0.0625-0.25 mg; 0.5-2.0 (.625 mg) = as long as the dose or dosage form value is lower than the standard medical dose or standard medical dosage form 0.312 to 1.25 mg; This calculation may be applied to standard medical doses and / or standard medical dosage forms for any AHCM described herein. In certain embodiments, the value is lower than the medical value standard. In other embodiments, the value is higher than a medical value standard.

  In one embodiment, the dose of AHCM agent is calculated based on the severity of fibrosis in the tumor sample.

  In some embodiments, the dose of AHCM agent has no anti-tumor effect when administered alone, for example, a non-hypertensive dose that does not have a significant effect of inhibiting or preventing tumor growth or progression It can be. In some embodiments, the dose of AHCM agent may be comparable to or greater than the standard medical dose or standard medical dosage formulation used for antihypertensive or anti-heart failure, and when administered alone Does not have an anti-tumor effect, eg, does not have a significant effect of inhibiting or preventing tumor growth or progression.

Therapeutic Methods In one aspect, the present invention provides a hyperproliferative disorder (eg, cancer) by administering an AHCM agent to a subject alone or in combination with a therapeutic agent, eg, an anticancer agent described herein. ).

  As used herein, unless otherwise specified, the terms “treat”, “treating”, and “treatment” refer to undesirable physiological changes or disorders such as the development or spread of cancer. Refers to both therapeutic and prophylactic or preventative measures aimed at preventing or slowing (lowering). Beneficial or desired clinical outcome may include detectable or undetectable symptoms, reduced symptoms, reduced degree of disease, stable (ie, does not worsen) disease, Includes delay or slowing of disease progression, remission or alleviation of disease state, and remission (partial or complete). “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those who already have a condition or disorder, including those who tend to have a condition or disorder, or persons whose condition or disorder should be prevented.

  For example, in the case of treatment of cancer, in some embodiments, therapeutic treatment refers to a tumor following administration by a method described herein or administration of a pharmaceutical composition described herein. Increase or inhibition or reduction of progression. For example, tumor growth or progression is inhibited or reduced by at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% after treatment To do. In another embodiment, tumor growth or progression is inhibited or reduced by greater than 50%, eg, at least about 60% or at least about 70% after treatment. In one embodiment, the tumor growth or progression is at least about 80%, at least about 90%, or more compared to a control (eg, in the absence of a pharmaceutical composition described herein). As described above, it is inhibited or decreased.

  In another embodiment, therapeutic treatment refers to the alleviation of at least one symptom associated with cancer. A measurable decrease includes a statistically significant decrease in a measurable marker or symptom, such as a cancer biomarker, such as a serum / plasma cancer biomarker in a blood sample, after treatment. In one embodiment, the at least one cancer biomarker or symptom is alleviated by at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, or at least about 50%. . In another embodiment, the at least one cancer biomarker or symptom is reduced by more than 50%, such as at least about 60% or at least about 70%. In one embodiment, the at least one cancer biomarker or symptom is at least about 80%, at least about 90, as compared to a control (eg, in the absence of the pharmaceutical composition described herein). % Or more.

  As used herein, unless otherwise specified, the terms “prevent”, “preventing”, and “prevention” are performed before a patient begins to suffer from a re-enhancement of cancer, And / or attempts to inhibit or reduce the severity of the cancer.

  As used herein, unless otherwise specified, a “therapeutically effective amount” of a compound provides a therapeutic benefit in the treatment of a disorder (eg, cancer) or a disorder (eg, An amount sufficient to delay or minimize one or more symptoms associated with cancer. A therapeutically effective amount of a compound means an amount of the therapeutic agent that alone or in combination with other therapeutic agents provides a therapeutic benefit in the treatment or management of a disorder. The term “therapeutically effective amount” includes improving the overall treatment of a disorder (eg, cancer), reducing or avoiding its symptoms or causes, or the therapeutic efficacy of another therapeutic agent. Enhancing amounts can be included.

  As used herein, unless otherwise specified, a “prophylactically effective amount” of a compound refers to a disorder (eg, cancer regrowth or one or more symptoms associated with cancer). Sufficient to prevent or prevent recurrence. By prophylactically effective amount of a compound is meant the amount of the compound that alone, or in combination with other therapeutic agents, provides a prophylactic benefit in preventing a disorder. The term “prophylactically effective amount” can include an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent.

  As used herein, the term “patient” or “subject” refers to an animal, typically a human, that will or will be the subject of treatment, observation, and / or experimentation. (Ie men and women of any age group, eg pediatric patients (eg infants, children, adolescents) or adult patients (eg young adults, middle-aged adults or elderly adults) or primates (eg cynomolgus monkeys, rhesus monkeys) ); Commercially important mammals such as cows, pigs, horses, sheep, goats, cats, and / or dogs; and / or commercially important mammals such as chickens, ducks, ducks, and / or turkeys Other mammals such as birds, including birds, when the term is used in conjunction with administration of a compound or drug, the patient Treatment, observation, and / or methods described. Herein were subject to administration and / or pharmaceutical compositions can also be used to treat livestock or pets such as cats and dogs.

  As used herein, “cancer” and “tumor” are synonyms.

  As used herein, “cancer therapy” and “cancer treatment” are synonymous.

  As used herein, “chemotherapy”, “chemotherapeutic agent”, “chemotherapeutic agent”, and “anticancer agent” are synonymous.

  In some embodiments, the AHCM agent, alone or in combination, is the primary treatment for cancer, i.e. has not previously been administered another drug intended to treat the cancer Used in subjects.

  In other embodiments, the AHCM agent, alone or in combination, is a secondary treatment for cancer, i.e. has been previously administered another drug intended to treat cancer Used in subjects.

  In other embodiments, the AHCM agent, alone or in combination, is a tertiary or quaternary treatment for cancer, i.e., previously two or three other drugs intended to treat cancer Used in subjects who have been administered

  In some embodiments, the AHCM agent is administered to the subject before, during, and / or after radiation or surgical treatment of cancer.

  In some embodiments, the AHCM agent alone is in a subject who has not previously responded to at least one cancer treatment or anticancer agent, including at least 2, at least 3, or at least 4 cancer treatment or anticancer agents. Or in combination with a cancer treatment or anti-cancer agent. In such embodiments, the AHCM agent may be administered to a subject in combination with a cancer therapy or anticancer agent that the subject has not previously responded to, or a different cancer therapy or anticancer agent than the subject has been treated with. A combination may be administered to a subject.

  In other embodiments, the AHCM agent is administered as an adjunct therapy, i.e., a treatment in addition to the main therapy. In embodiments, the adjuvant effect of AHCM administered in combination with the primary treatment can be additive.

  In certain embodiments, the cancer is an epithelial, mesenchymal, or hematological malignancy. In certain embodiments, the cancer to be treated is a solid tumor (eg, carcinoid, carcinoma, or sarcoma), soft tissue tumor (eg, heme malignancy), and metastatic lesions, eg, disclosed herein. Any metastatic lesion of cancer. In one embodiment, the cancer being treated is a fibrotic or fibrogenic solid tumor, such as a tumor having one or more of limited tumor perfusion, compressed blood vessels, or fibrotic tumor stroma. It is. In one embodiment, the solid tumor is pancreatic cancer (eg, pancreatic adenocarcinoma), breast cancer, colorectal cancer, lung cancer (eg, small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC)), skin cancer, ovarian cancer. , Liver cancer, esophageal cancer, endometrial cancer, stomach cancer, head and neck cancer, kidney cancer, or prostate cancer.

  “Hyperproliferative cancer disease or hyperproliferative cancer disorder” means whether it is malignant or benign, including all transformed cells and tissues and all cancerous cells and tissues, It means the expansion and proliferation of all neoplastic cells. Hyperproliferative diseases or hyperproliferative disorders include, but are not limited to, precancerous lesions, abnormal cell growth, benign tumors, malignant tumors, and “cancer”.

  As used herein, the term “cancer”, “tumor”, or “tumor tissue” refers to an abnormal amount of tissue due to excessive cell division, in some cases, hyperproliferative. Refers to tissue containing cells that are expressing, overexpressing, or abnormally expressing cellular proteins. A cancer, tumor, or tumor tissue contains “tumor cells”, which are neoplastic cells that have abnormally increasing properties and have no useful bodily functions. Cancers, tumors, tumor tissues, and tumor cells can be benign or malignant. A cancer, tumor, or tumor tissue can also include “tumor-associated non-tumor cells”, eg, vascular cells that form blood vessels that supply the tumor or tumor tissue. Non-tumor cells can be induced by tumor cells to replicate and develop (eg, induction of angiogenesis in a tumor or tumor tissue).

  Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia, or lymphoid malignancy. More specific examples of such cancers are shown below and include: squamous cell carcinoma (eg, epithelial squamous cell carcinoma), small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and Lung cancer including lung squamous cell carcinoma, peritoneal cancer, hepatocellular carcinoma, gastric cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectum Cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, and head and neck cancer. The term “cancer” includes primary malignant cells or malignant tumors (eg, cells that have not migrated to a body part of the subject other than the site of the first malignant disease or tumor), and secondary malignant cells. Or malignant tumors (eg, those arising from malignant cells or metastasis of tumor cells, ie, migration to a secondary site different from the site of the original tumor).

  Other examples of cancer or malignancies include acute childhood lymphoblastic leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, adrenocortical carcinoma, adult (primary) hepatocellular carcinoma , Adult (primary) liver cancer, adult acute lymphocytic leukemia, adult acute myeloid leukemia, adult Hodgkin disease, adult Hodgkin lymphoma, adult lymphocytic leukemia, adult non-Hodgkin lymphoma, adult primary liver cancer, adult soft part Tissue sarcoma, AIDS-related lymphoma, AIDS-related malignancy, anal cancer, astrocytoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, renal pelvic and ureteral cancer, central nervous system (primary) Lymphoma, central nervous system lymphoma, cerebellar astrocytoma, cerebral astrocytoma, cervical cancer, childhood (primary) hepatocellular carcinoma, childhood (primary) liver cancer, childhood acute lymphoblastic leukemia, childhood acute Myelocytic leukemia Childhood brain stem glioma, childhood cerebellar astrocytoma, childhood cerebral astrocytoma, childhood extracranial germ cell tumor, childhood Hodgkin disease, childhood Hodgkin lymphoma, childhood hypothalamic visual glioma, childhood lymphoblastic leukemia Pediatric medulloblastoma, pediatric non-Hodgkin lymphoma, pediatric pineal supratentorial primitive neuroectodermal tumor, pediatric primary liver cancer, pediatric rhabdomyosarcoma, pediatric soft tissue sarcoma, pediatric visual hypothalamic glioma, Chronic lymphocytic leukemia, chronic myeloid leukemia, colon cancer, cutaneous T-cell lymphoma, islet cell carcinoma, endometrial cancer, ependymoma, epithelial cancer, esophageal cancer, Ewing sarcoma and related tumors, exocrine pancreatic cancer, extracranial Germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer, female breast cancer, Gaucher disease, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal tumor, germ cell tumor, gestational trophoblast tumor, hairy cell leukemia Head and neck cancer, hepatocellular carcinoma, Hodgkin disease, Hodgkin lymphoma, hypergammaglobulinemia, hypopharyngeal cancer, intestinal cancer, intraocular melanoma, islet cell carcinoma, islet cell carcinoma, Kaposi sarcoma, kidney cancer, laryngeal cancer, lip Oral cancer, liver cancer, lung cancer, lymphoproliferative disorder, macroglobulinemia, male breast cancer, malignant mesothelioma, malignant thymoma, medulloblastoma, melanoma, mesothelioma, metastatic squamous neck of unknown origin Cancer, metastatic primary squamous cervical cancer, metastatic squamous cervical cancer, multiple myeloma, multiple myeloma / plasma cell neoplasm, myelodysplastic syndrome, myelogenous leukemia, myelocytic leukemia, bone marrow Proliferative disorder, nasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma during pregnancy, non-melanoma skin cancer, non-small cell lung cancer, metastatic squamous cervical cancer of unknown primary, oropharyngeal cancer, bone / Malignant fibrous sarcoma, osteosarcoma / malignant fibrous histiocytoma, osteosarcoma / Malignant fibrous histiocytoma, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low grade tumor, pancreatic cancer, paraproteinemia, purpura, parathyroid cancer, penile cancer, pheochromocytoma, pituitary tumor, trait Cell neoplasm / multiple myeloma, primary central nervous system lymphoma, primary liver cancer, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvic and ureteral cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoidosis Sarcoma, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cervical cancer, gastric cancer, tent primitive neuroectodermal pineal tumor, T cell lymphoma, testicular cancer, thymoma, thyroid Cancer, transitional cell carcinoma of the renal pelvis and ureter, transitional epithelial renal pelvic and ureteral cancer, choriocarcinoma, ureteral renal pelvic cell carcinoma, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, hypothalamic glioma, visual tract Cancer, Waldenstrom macroglobulinemia, Will Scan tumors, as well as include any other hyperproliferative diseases in addition to neoplasms located in an organ system listed above, but not limited to.

  In other embodiments, the AHCM agents described herein above are used to treat hyperproliferative disorders, such as hyperproliferative connective tissue disorders (eg, hyperproliferative fibrotic diseases). In one embodiment, the hyperproliferative fibrotic disease is systemic or organ specific. Exemplary hyperproliferative fibrotic diseases include systemic disorders (eg, systemic sclerosis, multifocal fibrosis, scleroderma graft-versus-host disease in bone marrow transplant recipients, renal systemic fibrosis) , Scleroderma), and organ-specific disorders such as, but not limited to, lung, liver, heart, kidney, pancreas, skin, and other organ fibrosis.

  In other embodiments, the subject to be treated has a hyperproliferative hereditary disorder, eg, a hyperproliferative hereditary disorder selected from Marfan syndrome or Royce Dietz syndrome. Losartan has been shown to treat human Marfan syndrome, a connective tissue disorder caused by mutations in the gene encoding fibrillin 1, an extracellular matrix protein (Dietz, HC et al. (2010) New Engl J Med 363 (9): 852-863). Fibrin 1 constitutes a component of elastic tissue microfibers and many other connective tissues. Patients suffering from Marfan syndrome have vascular abnormalities such as an aortic aneurysm. Vascular disease can result in vascular rupture and death after childhood. Dietz et al. First found in mouse models of Marfan syndrome that excessive activation of latent TGFβ plays an important role in pathophysiology. They used losartan in affected mice and had a marked effect on improving vascular architecture and prevented the development of aortic aneurysms. They also used losartan to treat children with Marfan syndrome, demonstrating that the drug can significantly prevent the progression of aortic and muscular lesions. Aortic diseases other than Marfan syndrome may also benefit from the use of losartan. Thus, inhibition of local latent activation of TGFβ and reduction of circulating active TGFβ levels alters the delivery and efficacy of nanotherapeutic drugs in connective tissues other than collagen in the extracellular matrix of cancer tissue Can have an effect on ingredients.

  In other embodiments, the hyperproliferative disorder (eg, hyperproliferative fibrotic disorder) is chronic obstructive pulmonary disease, asthma, aortic aneurysm, radiation-induced fibrosis, skeletal muscle myopathy, diabetic nephropathy, and / or arthritis Is selected from one or more.

  Additional exemplary hyperproliferative disorders that can be treated by the methods and compositions of the present invention are disclosed in Sounni, N.E. et al. (2010) Diseases Models & Mechanisms 3: 317-332.

Combination therapy The AHCM agents described herein above are combined with one or more additional therapies, such as, for example, radiation therapy, surgery, to treat the cancers described herein, and It will be appreciated that / or can be administered in combination with one or more therapeutic agents.

  “In combination with” does not imply that therapeutics or therapeutic agents must be administered at the same time and / or formulated together for delivery, although these methods of delivery are within the scope of the present invention. Is within. The pharmaceutical composition can be administered simultaneously with, before, or after one or more other additional treatments or therapeutic agents. In general, each agent will be administered at a dose and / or time schedule determined for that agent. It will further be appreciated that additional therapeutic agents utilized in this combination may be administered together in a single composition or may be administered separately in different compositions. The particular combination utilized in the plan will take into account the compatibility of the pharmaceutical composition of the invention with additional agents having therapeutic activity and / or the desired therapeutic effect to be achieved. .

  In general, additional therapeutic agents utilized in combination are expected to be utilized at a level that does not exceed the level when they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.

  In one embodiment, AHCM and / or treatment (eg, cancer or hyperproliferative treatment) is an inhibitor of a profibrotic pathway (eg, a pathway that is dependent or independent of activation of TGFβ and / or CTGF). ("Profibrotic pathway inhibitor") in combination. In one embodiment, AHCM and / or cancer treatment is administered in combination with one or more of inhibitors of endothelin 1, PDGF, Wnt / β catenin, IGF-1, TNFα, and / or IL-4. Is done. In another embodiment, AHCM and / or cancer treatment is administered in combination with an inhibitor of endothelin 1 and / or PDGF. In other embodiments, AHCM and / or cancer treatment is chemokine receptor type 4 (CXCR4) (eg AMD3100, MSX-122); stroma-derived factor 1 (SDF-1) (eg tannic acid); hedgehog ( For example, GDC-0449, cyclopamine, or GANT58) is administered in combination with one or more inhibitors.

  In other embodiments, AHCM is administered in combination with a low molecular weight chemotherapeutic agent. Exemplary low molecular weight chemotherapeutic agents include 13-cis-retinoic acid (isotretinoin, ACCUTANE®), 2-CdA (2-chlorodeoxyadenosine, cladribine, LEUSTATIN ™), 5-azacytidine ( Azacitidine, VIDAZA (registered trademark), 5-fluorouracil (5-FU, fluorouracil, ADRUCIL (registered trademark)), 6-mercaptopurine (6-MP, mercaptopurine, PURINETHOL (registered trademark)), 6-TG (6 -Thioguanine, thioguanine, THIOGUANINE TABLOID (registered trademark)), abraxane (protein-bound paclitaxel), actinomycin D (dactinomycin, COSMEGEN (registered trademark)), alitretinoin (PANRETIN (registered trademark)), all-trans retinoic acid ( ATRA, tretinoin, VESANOID (registered trademark), altretamine (hexamethylmelamine, HMM, HEXALEN (registered trademark)) , Ametopterin (methotrexate, methotrexate sodium, MTX, TREXALL (trademark), RHEUMATREX (trademark)), amifostine (ETHYOL (trademark)), arabinosylcytosine (Ara-C, cytarabine, CYTOSAR-U (trademark)) , Arsenic trioxide (TRISENOX (registered trademark)), asparaginase (Erwinia L-asparaginase, L-asparaginase, ELSPAR (registered trademark), KIDROLASE (registered trademark)), BCNU (carmustine, BiCNU (registered trademark)), bendamustine (TREANDA (Registered trademark)), bexarotene (TARGRETIN (registered trademark)), bleomycin (BLENOXANE (registered trademark)), busulfan (BUSULFEX (registered trademark), MYLERAN (registered trademark)), leucovorin calcium (citrobolum factor, folinic acid, leucovorin) , Camptothecin 11 (CPT-11, Irinotecan, CAMPTOSAR (registration Mark)), capecitabine (XELODA (registered trademark)), carboplatin (PARAPLATIN (registered trademark)), carmustine wafer (carmustine-containing polyfeprosan 20 implant, GLIADEL (registered trademark) wafer), CCI-779 (temsirolimus, TORISEL ( (Registered trademark)), CCNU (lomustine, CeeNU), CDDP (cisplatin, PLATINOL (registered trademark), PLATINOL-AQ (registered trademark)), chlorambucil (leukeran), cyclophosphamide (CYTOXAN (registered trademark), NEOSAR (registered trademark) Trademark)), dacarbazine (DIC, DTIC, imidazole carboxamide, DTIC-DOME (registered trademark)), daunomycin (daunorubicin, daunorubicin hydrochloride, rubidomycin hydrochloride, CERUBIDINE (registered trademark)), decitabine (DACOGEN (registered trademark)), Dexrazoxane (ZINECARD (registered trademark)), DHAD (mitoxantrone, NOVANTRONE Registered trademark), docetaxel (TAXOTERE (registered trademark)), doxorubicin (ADRIAMYCIN (registered trademark), RUBEX (registered trademark)), epirubicin (ELLENCE (registered trademark)), estramustine (EMCYT (registered trademark)), etoposide ( VP-16, etoposide phosphate, TOPOSAR (registered trademark), VEPESID (registered trademark), ETOPOPHOS (registered trademark), floxuridine (FUDR (registered trademark)), fludarabine (FLUDARA (registered trademark)), fluorouracil (cream) ) (CARAC ™, EFUDEX ™, FLUOROPLEX ™), gemcitabine (GEMZAR ™), hydroxyurea (HYDREA ™, DROXIA ™, MYLOCEL ™), idarubicin (IDAMYCIN (registered trademark)), ifosfamide (IFEX (registered trademark)), ixabepilone (IXEMPRA (registered trademark)), LCR (leulocristin, vincristine, VCR ONCOVIN (registered trademark), VINCASAR PFS (registered trademark), L-PAM (L-sarcorycin, melphalan, phenylalanine mustard, ALKERAN (registered trademark)), mechloretamine (mechloretamine hydrochloride, mustine, nitrogen mustard, MUSTARGEN (registered) Trademark)), mesna (MESNEX (trademark)), mitomycin (mitomycin C, MTC, MUTAMYCIN (trademark)), nelarabine (ARRANON (trademark)), oxaliplatin (ELOXATIN (trademark)), paclitaxel (TAXOL (trademark) ), ONXAL (trademark)), pegasparagase (PEG-L-asparaginase, ONCOSPAR (trademark)), PEMETREXED (ALIMTA (trademark)), pentostatin (NIPENT (trademark)), procarbazine (MATULANE (trademark) )), Streptozocin (ZANOSAR (registered trademark)), temozolomide (TEMODAR (registered trademark)), tenipo SID (VM-26, VUMON (registered trademark)), TESPA (thiophosphoamide, thiotepa, TSPA, THIOPLEX (registered trademark)), topotecan (HYCAMTIN (registered trademark)), vinblastine (vinblastine sulfate, vincaloicoblastine, VLB, ALKABAN-AQ®, VELBAN®, vinorelbine (vinorelbine tartrate, NAVELBINE®), and vorinostat (ZOLINZA®) are included, but are not limited to these.

  In another embodiment, the AHCM agent is administered with the biologic. Biologics useful in the treatment of cancer are known in the art, and binding molecules of the invention can be administered, for example, with such known biologics.

  For example, the FDA has approved the following biologics for the treatment of breast cancer: HERCEPTIN® (Trastuzumab (Genentech Inc., South San Francisco, Calif.); Human with antitumor activity in HER2-positive breast cancer Monoclonal antibody); FASLODEX® (fulvestrant (AstraZeneca Pharmaceuticals, LP, Wilmington, Del.); Estrogen receptor antagonist used to treat breast cancer); ARIMIDEX® (anastrozole) (AstraZeneca Pharmaceuticals, LP); a non-steroidal aromatase inhibitor that inhibits the enzyme aromatase required to make estrogen; Aromasin® (exemestane (Pfizer Inc., New York, NY); Irreversible steroidal aromatase inactivator used in treatment; FEMARA® (Letrazol (Novartis Pharmaceuti cals, East Hanover, NJ); non-steroidal aromatase inhibitor approved by the FDA to treat breast cancer; and NOLVADEX® (AstraZeneca Pharmaceuticals, LP); approved by the FDA to treat breast cancer Non-steroidal anti-estrogen). Other biologics with which the binding molecules of the invention can be combined include: AVASTIN® (Bevacizumab (Genentech Inc.); the first FDA-approved treatment designed to inhibit angiogenesis); ZEVALIN® (Biogen Idec, Cambridge, Mass .; a radiolabeled monoclonal antibody currently approved for the treatment of B-cell lymphoma).

  In addition, the FDA has approved the following biologics for the treatment of colorectal cancer: AVASTIN®; ERBITUX® (cetuximab (ImClone Systems Inc., New York, NY and Bristol-Myers Squibb, New York, NY); monoclonal antibody to epidermal growth factor receptor (EGFR); GLEEVEC® (imatinib mesylate; protein kinase inhibitor); and ERGAMISOL® (levamisole hydrochloride (Janssen Pharmaceutica Products, LP, Titusville, NJ); an immunomodulator approved by the FDA in 1990 as an adjunct combined with 5-fluorouracil after surgical resection in patients with Dukes' stage C colon cancer.

  Exemplary biologics for the treatment of lung cancer include TARCEVA® (erlotinib HCL (OSI Pharmaceuticals Inc., Melville, NY); to target the human epidermal growth factor receptor 1 (HER1) pathway Designed small molecules).

  Exemplary biologics for the treatment of multiple myeloma include VELCADE® Velcade (Bortezomib (Millennium Pharmaceuticals, Cambridge Mass.); A proteasome inhibitor). Additional biologics are thought to have multiple actions, including THALIDOMID® (thalidomide (Clegene Corporation, Warren, NJ); ability to inhibit myeloma cell growth and survival and anti-angiogenesis Immunomodulators).

  Additional exemplary cancer therapeutic antibodies include 3F8, abagovomab, adecatumumab, afutuzumab, alacizumab pegol, alemtuzumab (CAMPATH®, MABCAMPATH®) , Altumomab pentetate (HYBRI-CEAKER®), anatumomab mafenatox, anrukinzumab (IMA-638), apolizumab, alcisumomab (CEA-SCAN (registered) Trademark)), bavituximab, bectumomab (LYMPHOSCAN (registered trademark)), belimumab (BENLYSTA (registered trademark), LYMPHOSTAT-B (registered trademark)), besilesomab (SCINTIMUN (registered trademark)), bevacizumab (AVASTIN) (Registered trademark), bivatuzumab mertansine, blinatumom ab), brentuximab vedotin, cantuzumab mertansine, capromab pendetide (PROSTASCINT (registered trademark)), kazumaxomab (REMOVAB (registered trademark)), CC49, cetuximab (C225, ERBITUX (registered trademark)) , Citatuzumab bogatox, cixutumumab, clivatuzumab tetraxetan, conatumumab, dacetuzumab, denosumab (PROLIA), detumumab Ecromeximab (ecromeximab), edrecolomab (PANOREX (registered trademark)), erotuzumab (elotuzumab), epitumomab cituxetan (eptumuzumab, ertumaxomab (ertumaxomab) (REXOMUN (rac)) Figitumumab, Fresolim Bu, gariximab, gemtuzumab ozogamicin (MYLOTARG®), girentuximab, grammbatumumab vedotin, ibritumomab (ibritumomab tiuxetane, ZEVALIN®) ), Igovomab (INDIMACIS-125 (registered trademark)), inetumumab (intetumumab), inotuzumab ozogamicin, ipilimumab, iratumumab (iratumumab), ravetuzumab (CEA-CIDE (registered trademark)), lexatumumab, lintuzumab, , Lucatumumab (lucatumumab), lumiliximab, mapatumumab, matuzumab, miratuzumab, minretumomab (minretumomab), mitumomab (mitumomab), nacolomab fenatox (nacolomab tafenatobum) ERACIM (registered trademark), THERALOC (registered trademark), nofetumomab merpentan (VERLUMA (registered trademark)), ofatumumab (ARZERRA (registered trademark)), olaratumab (olaratumab), opoltuzumab monato Oportuzumab monatox, Oregobomab (OVAREX®), panitumumab (VECTIBIX®), pemtumomab (THERAGYN®), pertuzumab (OMNITARG®), pintumomab, pintumomab Pritumumab, ramcilmab, ranibizumab (LUCENTIS®), rilotumumab, rituximab (MABTHERA®, RITUXAN®), robatumumab (robatumumab), satumomab dezumado ), Siltuximab, sontuzumab (son tuzumab), tacatuzumab tetraxetane (AFP-CIDE (registered trademark)), taplitumomab paptox, tenatumomab, TGN1412, ticilimumab (tremelimumab), TN (BEXXAR®), trastuzumab (HERCEPTIN®), tremelimumab, tucotuzumab celmoleukin, veltuzumab, borociximab, votumumab (HUMASPECT®), salutumumab (HUMAX-EGFR) (Registered trademark)), and zanolimumab (HUMAX-CD4®).

  In other embodiments, AHCM is administered in combination with a viral cancer therapeutic agent. Exemplary viral cancer therapeutic agents include vaccinia virus (vvDD-CDSR), carcinoembryonic antigen-expressing measles virus, recombinant vaccinia virus (TK deletion + GM-CSF), Seneca Valley virus 001, Newcastle disease virus Coxsackievirus A21, GL-ONC1, EBNA1 C-terminal / LMP2 chimeric protein expressing recombinant modified vaccinia ankara vaccine, carcinoembryonic antigen expressing measles virus, G207 tumor regression virus, p53 expression modified vaccinia ankara virus vaccine, OncoVEX GM-CSF Modified type 1 herpes simplex virus, fowlpox virus vaccine vector, recombinant vaccinia prostate specific antigen vaccine, human papillomavirus 16/18 L1 virus-like particle / AS04 vaccine, MVA-EBNA1 / LMP2 Inj. Vaccine, tetravalent HPV vaccine, four -Valent human papillomavirus (6, 11, 16, 18) recombinant vaccine (GARDA SIL (registered trademark)), recombinant fowlpox-CEA (6D) / TRICOM vaccine; recombinant vaccinia-CEA (6D) -TRICOM vaccine, recombinant modified vaccinia Ankara-5T4 vaccine, recombinant fowlpox-TRICOM vaccine, Oncolytic herpesvirus NV1020, HPV L1 VLP vaccine V504, human papillomavirus bivalent (type 16 and 18) vaccine (CERVARIX®), herpes simplex virus HF10, Ad5CMV-p53 gene, recombinant vaccinia DF3 / MUC1 vaccine, Recombinant vaccinia-MUC-1 vaccine, recombinant vaccinia-TRICOM vaccine, ALVAC MART-1 vaccine, human preproenkephalin (NP2) expression replication deficiency type I herpes simplex virus (HSV-1) vector, wild type reovirus, reovirus Recombinant modified vaccinia encoding type 3 Dearing strain (REOLYSIN (registered trademark)), tumor regression virus HSV1716, Epstein-Barr virus target antigen Ankara (MVA) based vaccine, recombinant fowlpox-prostate specific antigen vaccine, recombinant vaccinia prostate specific antigen vaccine, recombinant vaccinia-B7.1 vaccine, rAd-p53 gene, Ad5-delta 24RGD, HPV vaccine 580299, JX -594 (Thymidine kinase-deficient vaccinia virus + GM-CSF), HPV-16 / 18 L1 / AS04, fowlpox virus vaccine vector, vaccinia tyrosinase vaccine, MEDI-517 HPV-16 / 18 VLP AS04 vaccine, herpes simplex virus TK99UN Adenoviral vector containing thymidine kinase, HspE7, FP253 / fludarabine, ALVAC (2) Melanoma multiple antigen therapy vaccine, ALVAC-hB7.1, Canarypox-hIL-12 melanoma vaccine, Ad-REIC / Dkk -3, rAd-IFN SCH 721015, TIL-Ad-INFg, Ad-ISF35, and Coxsackievirus A21 (CVA21, CAVATAK®), but are not limited thereto.

  In other embodiments, AHCM is administered in combination with a nanopharmaceutical. Exemplary cancer nanopharmaceuticals include ABRAXANE® (paclitaxel-conjugated albumin nanoparticles), CRLX101 (CPT conjugated to a linear cyclodextrin-based polymer), CRLX288 (docetaxel and biodegradable polymer poly ( Conjugates with lactate glycolic acid), liposomal cytarabine (liposome Ara-C, DEPOCYT ™), liposomal daunorubicin (DAUNOXOME ™), liposomal doxorubicin (DOXIL ™, CAELYX ™), Daunorubicin citrate encapsulated liposomes (DAUNOXOME®), and PEG anti-VEGF aptamer (MACUGEN®) are included, but are not limited to these.

  In some embodiments, the AHCM agent is administered in combination with paclitaxel or a paclitaxel formulation, eg, TAXOL®, protein-bound paclitaxel (eg, ABRAXANE®). Exemplary paclitaxel formulations include nanoparticulate albumin-conjugated paclitaxel (ABRAXANE® sold by Abraxis Bioscience), docosahexaenoic acid-conjugated paclitaxel (DHA-paclitaxel, Taxoprexin sold by Protarga), polyglutamic acid-conjugated paclitaxel (PG-paclitaxel, paclitaxel polygourex, CT-2103, XYOTAX sold by Cell Therapeutic), tumor-activated prodrug (TAP), ANG105 (three sold by ImmunoGen) Angiopep-2 bound to paclitaxel molecule), paclitaxel-EC-1 (see paclitaxel bound to erbB2 recognition peptide EC-1; see Li et al, Biopolymers (2007) 87: 225-230), and glucose conjugated Gated paclitaxel (eg, 2'-paclitaxel methyl 2-glucopyranos . Succinate, Liu et al, Bioorganic & Medicinal Chemistry Letters (2007) 17: 617-620 see) include, but are not limited to these.

  Exemplary RNAi and antisense RNA agents for treating cancer include, but are not limited to, CALAA-01, siG12D LODER (Local Drug EluteR), and ALN-VSP02.

  Other cancer therapeutic agents include cytokines (eg, aldesleukin (IL-2, interleukin 2, PROLEUKIN (registered trademark)), α interferon (IFNα, interferon α, INTRON (registered trademark) A (interferon α-2b) , ROFERON-A (registered trademark) (interferon α-2a)), epoetin alfa (PROCRIT (registered trademark)), filgrastim (G-CSF, granulocyte colony stimulating factor, NEUPOGEN (registered trademark)), GM-CSF (Granulocyte macrophage colony-stimulating factor, salgramostim, LEUKINE (trademark)), IL-11 (interleukin 11, oprel bekin, NEUMEGA (trademark)), interferon α-2b (PEG conjugate) (PEG interferon, PEG-INTRON ( Trademark)), and pegfilgrastim (NEULASTA ™)), hormone therapeutic agents (eg, aminoglutethimide (CYTADREN ™) )), Anastrozole (ARIMIDEX (trademark)), bicalutamide (CASODEX (trademark)), exemestane (AROMASIN (trademark)), fluoxymesterone (HALOTESTIN (trademark)), flutamide (EULEXIN (trademark)) ), Fulvestrant (FASLODEX (registered trademark)), goserelin (ZOLADEX (registered trademark)), letrozole (FEMARA (registered trademark)), leuprolide (ELIGARD (registered trademark), LUPRON (registered trademark), LUPRON DEPOT (registered trademark) ), VIADUR (trademark), megestrol (megestrol acetate, MEGACE (trademark)), nilutamide (ANANDRON (trademark), NILANDRON (trademark)), octreotide (octreotide acetate, SANDOSTATIN (trademark) ), SANDOSTATIN LAR (registered trademark)), Raloxifene (EVISTA (registered trademark)), Lomiprostim (NPLATE (registered trademark)), Tamoxif (NOVALDEX®), and toremifene (FARESTON®)), phospholipase A2 inhibitors (eg, anagrelide (AGRYLIN®)), biological response modifiers (eg, BCG (THERACYS ( Registered trademark), TICE®) and darbepoetin alfa (ARANESP®)), targeted therapeutic agents (eg, bortezomib (VELCADE®), dasatinib (SPRYCEL ™), Denileukin diftitox (ONTAK (registered trademark)), erlotinib (TARCEVA (registered trademark)), everolimus (AFINITOR (registered trademark)), gefitinib (IRESSA (registered trademark)), imatinib mesylate (STI-571, GLEEVEC (registered trademark)), Lapatinib (TYKERB®), Sorafenib (NEXAVAR®), and SU11248 (Sunitinib, SUTENT®)), an immunomodulating anti-vascular shape Agents (eg CC-5013 (lenalidomide, REVLIMID®) and thalidomide (THALOMID®)), glucocorticoids (eg cortisone (hydrocortisone, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, ALA -CORT (registered trademark), HYDROCORT ACETATE (registered trademark), hydrocortisone phosphate LANACORT (registered trademark), SOLU-CORTEF (registered trademark), decadron (dexamethasone, dexamethasone acetate, dexamethasone phosphate sodium, DEXASONE (registered) Trademark), DIODEX (registered trademark), HEXADROL (registered trademark), MAXIDEX (registered trademark)), methylprednisolone (6-methylprednisolone, methylprednisolone acetate, methylprednisolone succinate sodium, DURALONE (registered trademark) Trademark), MEDRALONE (registered trademark), MEDROL (registered trademark), M-PREDNISOL (registered trademark), SOLU-MEDROL (registered trademark)), prednisolone (DELTA-CORTEF (registered trademark), ORAPRED (registered trademark), PEDIAPRED ( Registered trademark), PRELONE®), and prednisone (DELTASONE®, LIQUID PRED®, METICORTEN®, ORASONE®)), and bisphosphonates (eg, pamidronic acid (AREDIA)) (Registered trademark)) and zoledronic acid (ZOMETA (registered trademark))).

  In some embodiments, AHCM agents are used in combination with tyrosine kinase inhibitors (eg, receptor tyrosine kinase (RTK) inhibitors). Exemplary tyrosine kinase inhibitors include epidermal growth factor (EGF) pathway inhibitors (eg, epidermal growth factor receptor (EGFR) inhibitors), vascular endothelial growth factor (VEGF) pathway inhibitors (eg, vascular endothelial growth). Factor receptor (VEGFR) inhibitor (eg, VEGFR-1 inhibitor, VEGFR-2 inhibitor, VEGFR-3 inhibitor)), platelet derived growth factor (PDGF) pathway inhibitor (eg, platelet derived growth factor receptor) (PDGFR) inhibitors (eg, PDGFRβ inhibitors)), RAF-1 inhibitors, KIT inhibitors, and RET inhibitors, but are not limited to these. In some embodiments, the anti-cancer agent used in combination with the AHCM agent is axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTIN ™, AZD2171), dasatinib (SPRYCEL®, BMS-354825 ), Erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®)), Restaurtinib (CEP-701), Neratinib (HKI-272), Nilotinib (TASIGNA (registered trademark)), Semaxanib (cemaxinib, SU5416), Sunitinib (SUTENT (registered trademark), SU11248), Toceranib (PALLADIA (registered trademark) )), Vandetanib (ZACTIMA®, ZD6474), batalanib (PTK787, PTK / ZK), trastuzumab (HERCEPTIN®), bevacizumab ( AVASTIN (R)), Rituximab (RITUXAN (R)), Cetuximab (ERBITUX (R)), Panitumumab (VECTIBIX (R)), Ranibizumab (Lucentis (R)), Nilotinib (TASIGNA (R)) ), Sorafenib (NEXAVAR®), alemtuzumab (CAMPATH®), gemtuzumab ozogamicin (MYLOTARG®), ENMD-2076, PCI-32765, AC220, dovitinibactate (dovitinib lactate) (TKI258, CHIR-258), BIBW 2992 (TOVOK (trademark)), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF (registered trade mark) )), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154, CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, XL228 , AEE788, AG-490, AST-6, BMS-599626, CUDC-101, PD153035, peritinib (EKB- 569), vandetanib (zactima), WZ3146, WZ4002, WZ8040, ABT-869 (linifanib), AEE788, AP24534 (ponatinib), AV-951 (tibozanib), axitinib, BAY 73-4506 (regorafenib) , Brivanib alaninate (BMS-582664), brivanib (BMS-540215), cediranib (AZD2171), CHIR-258 (dobitinib), CP 673451, CYC116, E7080, Ki8751, masitinib (AB1010), MGCD-265 , Motesanib diphosphate (AMG-706), MP-470, OSI-930, pazopanib hydrochloride, PD173074, sorafenib tosylate (Bay 43-9006), SU 5402, TSU-68 (SU6668), Selected from the group consisting of Batalanib, XL880 (GSK1363089, EXEL-2880). The selected tyrosine kinase inhibitor is selected from sunitinib, erlotinib, gefitinib, or sorafenib. In one embodiment, the tyrosine kinase inhibitor is sunitinib.

  In one embodiment, AHCM is administered in combination with one or more of an anti-angiogenic agent or a vascular targeting agent or a vascular disrupting agent. Exemplary anti-angiogenic agents include, among others, VEGF inhibitors (eg, anti-VEGF antibodies (eg, bevacizumab); VEGF receptor inhibitors (eg, itraconazole); inhibitors of endothelial cell proliferation and / or migration) (Eg, carboxamidotriazole, TNP-470); including but not limited to inhibitors of angiogenesis stimulators (eg, suramin) Vascular targeting agents (VTA) or vascular disrupting agents (VDA) Designed to damage the tumor vasculature (blood vessels) and cause central necrosis (eg, outlined in Thorpe, PE (2004) Clin. Cancer Res. Vol. 10: 415-427). Exemplary small molecule VTAs include drugs that destabilize microtubules (eg, combretastatin A-4 disodium phosphate (CA4P), ZD6126, AVE8062, Oxi 4503); (Vadimezan) (ASA404) included However, it is not limited to these.

It is recognized that isotope-labeled anti-tumor antibodies have been used successfully in animal models, in some cases in humans, to destroy cells in solid tumors and lymphoma / leukemia Will. Exemplary radioisotopes include ⁹⁰ Y, ¹²⁵ I, ¹³¹ I, ¹²³ I, ¹¹¹ In, ¹⁰⁵ Rh, ¹⁵³ Sm, ⁶⁷ Cu, ⁶⁷ Ga, ¹⁶⁶ Ho, ¹⁷⁷ Lu, ¹⁸⁶ Re, and ¹⁸⁸ Re It is. Radionuclides act by causing ionizing radiation that causes multiple strand breaks in the nuclear DNA and leads to cell death. The isotopes used to make therapeutic conjugates typically yield high energy alpha or beta particles with a short range. Such radionuclides kill neighboring cells, eg, neoplastic cells with attached or invaded conjugates. They have little or no effect on non-localized cells. Radionuclides are essentially non-immunogenic.

It will also be appreciated that, in accordance with the teachings herein, binding molecules can be conjugated to different radiolabels for diagnostic and therapeutic purposes. For this purpose, the above-mentioned US Pat. Nos. 6,682,134, 6,399,061, and 5,843,439 are radiolabeled therapeutic conjugates for diagnostic “imaging” of tumors prior to administration of therapeutic antibodies. The gate is disclosed. The “In2B8” conjugate is a bifunctional chelator, namely MX-DTPA comprising a 1: 1 mixture of 1-isothiocyanatobenzyl-3-methyl-DTPA and 1-methyl-3-isothiocyanatobenzyl-DTPA. Contains mouse monoclonal antibody 2B8 specific for human CD20 antigen attached to ¹¹¹ In via (diethylenetriaminepentaacetic acid). ¹¹¹ In can be safely administered from about 1 to about 10 mCi without detectable toxicity; imaging data is particularly preferred as a diagnostic radionuclide because it generally predicts subsequent dispersion of ⁹⁰ Y-labeled antibodies . Most imaging studies utilize antibodies labeled with 5mCi ¹¹¹ In because this dose is safe and has increased imaging efficiency compared to lower doses. Optimal imaging occurs 3-6 days after antibody administration. See, for example, Murray, J. Nuc. Med. 26: 3328 (1985) and Carraguillo et al., J. Nuc. Med. 26:67 (1985).

  In certain embodiments, the AHCM agent and the additional anticancer agent are administered simultaneously (eg, administration of the two agents at the same time or on the same day, or within the same treatment regime), and / or sequentially. (E.g. administration of one agent in a period of time followed by administration of other agents in a second period or within a different treatment regime).

  In one embodiment, AHCM is administered before the anticancer agent. In other embodiments, AHCM is administered prior to the anticancer agent, followed by co-administration of AHCM and the anticancer agent.

  In certain embodiments, the AHCM agent and the additional anticancer agent are administered simultaneously. For example, in certain embodiments, the AHCM agent and the additional anticancer agent are administered simultaneously, on the same day, or within the same treatment regime. In certain embodiments, the AHCM agent is administered prior to the additional anticancer agent on the same day or within the same treatment regime.

  In certain embodiments, the AHCM agent is administered at the same time as the additional anticancer agent for a period of time after which treatment with the additional anticancer agent is discontinued and treatment with the AHCM agent is continued.

  In other embodiments, the AHCM agent is administered at the same time as the additional anticancer agent for a period of time after which treatment with the AHCM agent is discontinued and treatment with the additional anticancer agent is continued.

  In certain embodiments, the AHCM agent and the additional anticancer agent are administered sequentially. For example, in certain embodiments, the AHCM agent is administered after the additional anticancer drug treatment regime is completed. In certain embodiments, the additional anticancer agent is administered after the AHCM agent treatment regime is completed.

  In some embodiments, AHCM agents and anticancer agents can be administered in pulsed administration. In other embodiments, they may be administered as pulse chase administration, e.g., an AHCM agent is administered over a short period (pulse) followed by an anti-cancer agent over a longer period (e.g. chase), Or vice versa.

Diagnostic methods and assays
AHCM agents can be used to improve the diagnosis, treatment, prevention, and / or prognosis of cancer in a mammal, preferably a human. These diagnostic assays may be performed in vivo or in vitro, for example, on blood samples, biopsy tissue, or autopsy tissue.

  Accordingly, the present invention provides a process for measuring the expression level of a target protein or target transcript in tissue or other cells or body fluids derived from an individual, and the measured expression level as a standard target expression level in normal tissues or body fluids. Provides a diagnostic method useful in the diagnosis of cancer, comprising a step wherein an increase in expression level relative to a standard is indicative of a disorder.

  One embodiment comprises assaying for expression of a target in an individual tissue sample or body fluid sample and comparing the presence or level of target expression in the sample to the presence or level of target expression in a panel of standard tissue samples or standard fluid samples. An abnormal hyperproliferative cell in a liquid sample or tissue sample, for example pre-cancerous, wherein the detection of increased target expression relative to the target expression or standard is indicative of abnormal hyperproliferative cell increase A method of detecting the presence of a cell or cancer cell is provided.

  One aspect of the invention is a method for in vivo detection or diagnosis of cancer in a subject, preferably a mammal, most preferably a human. In one embodiment, the diagnosis comprises (a) subject a subject who has been or has been treated with AHCM with an effective amount (eg, parenteral, subcutaneous) of a labeled antibody or fragment thereof against a cancer antigen. Or (intraperitoneally) administering; (b) the labeled antibody is preferentially concentrated at the site of interest where the target is expressed (and unbound labeled molecules are brought to background levels) (C) determining the background level; and (d) detecting the labeled molecule in the subject and labeling above the background level. The detection of the detected molecule should indicate that the subject has a specific disease or disorder associated with abnormal expression of the target. The background level can be determined by a variety of methods, including comparing the amount of labeled molecule detected to a previously determined standard value for a particular system.

It will be understood in the art that the size of the object and the imaging system used will determine the amount of imaging moiety needed to produce a diagnostic image. In the case of a radioisotope moiety for human subjects, the amount of radioactivity injected will typically be in the range of about 5-20 millicuries, for example ⁹⁹ Tc. The labeled binding molecule, eg, antibody or antibody fragment, will then preferentially accumulate at the location of the cell containing the particular protein. In vivo tumor imaging is described in SWBurchiel et al., “Immunopharmacokinetics of Radiolabeled Antibodies and Their Fragments.” (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, SWBurchiel and BARhodes, eds., Masson Publishing Inc. (1982). Has been.

  Depending on a number of variables, including the type of label used and the mode of administration, the labeled molecules are preferentially concentrated to sites in the subject and unbound labeled molecules are removed to background levels. The interval after administration to make it possible is 6 to 48 hours or 6 to 24 hours or 6 to 12 hours. In another embodiment, the post-administration interval is 5-20 days or 7-10 days.

  The presence of the labeled molecule can be detected in the patient using methods known in the art for in vivo scanning. These methods depend on the type of label used. One skilled in the art will be able to determine an appropriate method for detecting a particular label. Methods and devices that can be used in the diagnostic methods of the present invention include computed tomography (CT), positron emission tomography (PET), magnetic resonance imaging (MRI), and whole body scans such as ultrasonography, X-rays This includes but is not limited to examination, nuclear magnetic resonance imaging (NMR), CAT scan, or electron spin resonance imaging (ESR).

Pharmaceutical Compositions The compositions described herein can be incorporated into a variety of formulations for administration. More specifically, the composition can be formulated into a pharmaceutical composition by combining with a suitable pharmaceutically acceptable carrier or diluent, such as a capsule, powder, granule, gel, slurry, ointment, It can be formulated into preparations in semisolid, liquid, or gaseous form, such as solutions, suppositories, injections, inhalants, and aerosols. Thus, administration of the composition can be accomplished in a variety of ways, including oral administration, buccal administration, rectal administration, parenteral administration, intraperitoneal administration, intradermal administration, transdermal administration, intratracheal administration. Further, the compositions may be administered locally rather than systemically in a depot or sustained release formulation.

  In addition, the compositions may be formulated with common excipients, diluents, or carriers, compressed into tablets, or formulated as elixirs or solutions for convenient oral administration. Or may be administered by intramuscular or intravenous routes. The composition may be administered transdermally and may be formulated as a sustained release dosage form or the like. The compositions may be administered alone, in combination with each other, or may be used in combination with other known compounds (described herein).

  Suitable formulations for use in the present invention are found in Remington's Pharmaceutical Sciences (1985). See also Langer (1990) Science 249: 1527-1533 for a review of methods for drug delivery. The pharmaceutical compositions described herein can be manufactured in a manner known to those skilled in the art, for example, by mixing, dissolving, granulating, dragee making, grinding, emulsifying, encapsulating, capturing, or lyophilizing processes. . The following methods and excipients are merely exemplary and are in no way limiting.

  Compositions for oral administration can be formulated by combining with pharmaceutically acceptable carriers known in the art. Such carriers can be formulated as pills, capsules, emulsions, lipophilic and hydrophilic suspensions, liquids, gels, syrups, slurries, suspensions, etc. for ingestion by the patient being treated. Can be formulated. Pharmaceutical preparations for oral use include mixing the composition with excipients and, if desired, adding appropriate auxiliaries, then processing the granule mixture to obtain tablets or dragee cores. Can be obtained. Suitable excipients are in particular bulking agents such as sugar containing lactose, sucrose, mannitol or sorbitol; for example corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose Cellulose preparations such as sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP).

  For administration by inhalation, a composition for use in accordance with the present invention may be added by use of a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. Conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer, or from a dry powder inhaler without a propellant. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges containing a powder mixture of the compound and a suitable powder base such as lactose or starch can be formulated for use in an inhaler or insufflator.

  The composition may be formulated for parenteral administration by injection, eg, by bolus injection or continuous infusion. Formulations for injection can be presented in unit dosage form, for example, ampoules or multi-dose containers with an added preservative. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous medium, with formulation agents such as suspending, stabilizing, and / or dispersing agents. May be contained.

  The composition contains, for example, a conventional suppository base such as cocoa butter, carbowax, polyethylene glycol, or other glycerides that melts at body temperature but becomes solid at room temperature. Or it may be formulated into a rectal composition such as a retention enema.

  In addition, the composition may be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compound may be formulated using an appropriate polymeric or hydrophobic material (eg, as an emulsion with an acceptable oil) or formulated using an ion exchange resin. Alternatively, it may be formulated as a poorly soluble derivative, for example, as a poorly soluble salt.

  Lipid particles (eg, liposomes) and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Liposomes that stay in circulation for a long time, such as stealth liposomes, can be used. Such liposomes are generally described in US Pat. No. 5,013,556. The compounds of the present invention may be administered by controlled release means and / or delivery devices such as those described in US Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719.

  Suitable pharmaceutical compositions for use in the present invention include those containing an active ingredient in a therapeutically effective amount. The amount of composition administered will, of course, be dependent on the subject being treated, on the subject's weight, the severity of the affliction, the manner of administration and the judgment of the prescribing physician. Determination of an effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. In general, a suitable daily dose of an AHCM agent and / or cancer therapeutic agent can be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose may generally depend on the factors described above.

  Subjects that receive this treatment are any animal in need thereof, including primates, particularly humans, horses, cows, pigs, sheep, poultry, dogs, cats, mice, and rats.

  The compounds can be administered daily, every other day, three times a week, twice a week, every week, or every other week. The dosing schedule may include a “drug holiday”, ie the drug is administered 2 weeks on, 1 week off, or 3 weeks on, 1 week off, 4 weeks on, 1 week off, etc. Or may be administered continuously without a drug holiday. The compounds can be administered orally, intravenously, intraperitoneally, externally, transdermally, intramuscularly, subcutaneously, intranasally, sublingually, or by any other route.

  Because AHCM agents are administered in combination with other treatments (such as additional chemotherapeutic drugs, radiation, or surgery), the dose of each agent or each treatment is greater than the corresponding dose for single agent therapy Can be low. The determination of the mode of administration and the exact dosage is well within the knowledge of a skilled physician.

  In one embodiment, AHCM is administered via an implantable infusion device, such as a pump. Implantable infusion devices typically include a housing containing a liquid reservoir that can be percutaneously filled with a hypodermic needle that penetrates a fill port septum. Drug treatment reservoirs are generally coupled via an internal flow path to a device outlet port to deliver fluid through a catheter to a patient's body part. A typical infusion device also includes a controller and a liquid transfer mechanism, such as a pump or valve, to move liquid from the reservoir through the internal flow path to the outlet port of the device.

Nanoparticles The AHCM agents described herein, anticancer agents (eg, low molecular weight anticancer agents, medium molecular weight anticancer agents described herein), or both, may be packaged into nanoparticles.

  Typically, the nanoparticles have a diameter of 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, or 200 nm, or 200-1,000 nm, e.g., 10 nm, 15 nm, 20 nm, 25 nm 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, or 200 nm, or 20 nm or 30 nm or 50-400 nm. Smaller particles tend to be removed more quickly from the system. The drug may be entrapped within the nanoparticles, or may be coupled to the nanoparticles, eg, covalently coupled, or otherwise attached.

  Lipid or oil-based nanoparticles, such as liposomes and solid lipid nanoparticles, can be used to deliver the agents described herein. DOXIL® is an example of a liposome nanoparticle. Solid lipid nanoparticles for the delivery of anticancer drugs are described in Serpe et al. (2004) Eur. J. Pharm. Bioparm. 58: 673-680 and Lu et al. (2006) Eur. J. Pharm. Sci. 28: 86-95. Polymer-based nanoparticles, such as PLGA-based nanoparticles, can be used to deliver the agents described herein. They tend to rely on biodegradable scaffolds (with or without covalent bonding to the polymer) in which the therapeutic agent is inserted into the polymer matrix. PLGA is widely used in polymer nanoparticles (Hu et al. (2009) J. Control. Release 134: 55-61; Cheng et al. (2007) Biomaterials 28: 869-876, and Chan et al. (See 2009) Biomaterials 30: 1627-1634). Nanoparticles based on PEGylated PLGA can also be used to deliver anti-cancer agents (eg, Danhhier et al., (2009) J. Control. Release 133: 11-17, Gryparis et al (2007) Eur. J. Pharm. Biopharm. 67: 1-8). Metal-based, eg, gold-based nanoparticles, can also be used to deliver anticancer agents. Protein-based, eg, albumin-based nanoparticles, can be used to deliver the agents described herein, eg, the agent can be bound to human albumin nanoparticles. An exemplary anticancer agent / protein nanoparticle is Abraxane® in which paclitaxel is bound to albumin nanoparticles.

  Nanoparticles can utilize active targeting, passive targeting, or both. Active targeting can rely on the inclusion of a ligand that binds to a preselected site, eg, a target present at or near a solid tumor. Passive targeting nanoparticles can diffuse and accumulate at sites of interest, for example, sites characterized by excessively leaking microvasculature, such as found in tumors, and sites of inflammation .

  A wide range of nanoparticles are known in the art. Exemplary approaches include WO2010 / 005726, WO2010 / 005723, WO2010 / 005721, WO2010 / 121949, WO2010 / 0075072, WO2010 / 068866, WO2010 / 005740, WO2006 / 014626, 7,820,788, 7,780,984 (the contents of which are in their entirety) And those described in (incorporated herein by reference).

  The invention may be defined in any of the following numbered sections:

1. Based on the need for improved delivery or efficacy of cancer treatment, identifying the subject as requiring antihypertensive and / or receipt of a collagen modifying agent (“AHCM”); and (a) the subject A method of improving the delivery or efficacy of a cancer treatment in a subject, comprising either or both of the step of administering AHCM to: or (b) administering a cancer treatment comprising:
The method wherein the AHCM is administered at a dosage sufficient to improve the delivery or efficacy of the cancer treatment.

2. Identifying the subject as requiring antihypertensive and / or receipt of a collagen modifying agent (“AHCM”) based on the need for improved delivery or efficacy of cancer treatment; and (a) the subject A method of treating or preventing cancer in a subject, comprising either or both of: administering AHCM to: or (b) administering a cancer therapy comprising:
A method wherein AHCM is administered at a dosage sufficient to treat or prevent cancer.

3. (a) 1H, 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 45nm, 50nm, 75nm, 100nm, 150nm, larger than 200nm, but with a hydrodynamic diameter smaller than 300nm as AHCM, Cancer treatment, or both being administered;
(B) the subject has not been administered a dose of AHCM within 5 days, within 10 days, within 30 days, within 60 days, or within 100 days of the start of cancer diagnosis or AHCM medication;
(C) the subject is not hypertensive or hypertensive prior to administration of AHCM;
(D) AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier That;
(E) AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier And (f) AHCM is at least 1 hour, 5 hours, 10 hours, or 24 hours; at least 2 days, 5 days, 10 days, or 14 days; at least 2 weeks , 3 weeks, 4 weeks, 5 weeks, or 6 weeks; at least 2 months, 3 months, 4 months, 5 months, or 6 months; or a period of at least 1 year, 2 years, 3 years, 4 years, or 5 years The method of paragraph 1 or 2, comprising one or more of being administered continuously over time.

4.AHCM
(I) angiotensin II receptor blocker (AT ₁ blocker),
(Ii) renin angiotensin aldosterone antagonists (“RAAS antagonists”),
(Iii) angiotensin converting enzyme (ACE) inhibitor,
(Iv) thrombospondin 1 (TSP-1) inhibitor,
The method according to any one of Items 1 to 3, selected from one or more of (v) a transforming growth factor β1 (TGF-β1) inhibitor or (vi) a connective tissue growth factor (CTGF) inhibitor .

5. AHCM is losartan (COZAAR (registered trademark)), candesartan (ATACAND (registered trademark)), eprosartan mesylate (TEVETEN (registered trademark)), EXP 3174, irbesartan (AVAPRO (registered trademark)), L158,809 An AT ₁ inhibitor selected from one or more of: olmesartan (BENICAR®), salalasin, telmisartan (MICARDIS®), valsartan (DIOVAN®), or derivatives thereof Item 5. The method according to any one of Items 1 to 4.

  6. The method according to any one of Items 1 to 5, wherein AHCM is losartan.

  7. AHCM is from one or more of aliskiren (TEKTURNA®, RASILEZ®), remixylene (Ro 42-5892), enalkylene (A-64662), SPP635, or derivatives thereof Item 7. The method according to any one of Items 1 to 6, which is a selected RAAS antagonist.

  8. AHCM is Benazepril (LOTENSIN (registered trademark)), Captopril (CAPOTEN (registered trademark)), Enalapril (VASOTEC (registered trademark)), Hoshinopril (MONOPRIL (registered trademark)), Lisinopril (PRINIVIL (registered trademark)), ZESTRIL (Registered trademark)), moexipril (UNIVASC (registered trademark)), perindopril (ACEON (registered trademark)), quinapril (ACCUPRIL (registered trademark)), ramipril (ALTACE (registered trademark)), trandolapril (MAVIK (registered trademark)) )), Or the method according to any one of Items 1 to 7, which is an ACE inhibitor selected from one or more of these derivatives.

  9. The method according to any one of Items 1 to 8, wherein AHCM is a TSP-1 inhibitor selected from one or more of ABT-510, CVX-045, LSKL, or derivatives thereof.

  10. The method of paragraph 4, wherein the TGF-β1 inhibitor is selected from one or more of an anti-TGF-β1 antibody or a TGF-β1 peptide inhibitor.

  11. The method of paragraph 4, wherein the CTGF inhibitor is selected from one or more of DN-9693, FG-3019, or derivatives thereof.

  12. The method of any of paragraphs 1-11, wherein AHCM is administered in an amount sufficient to enhance the dispersion or efficacy of the cancer treatment.

  13. AHCM may reduce the level or production of collagen, decrease tumor fibrosis, increase stromal tumor transport, improve tumor perfusion, or enhance penetration or spread of cancer treatment in the tumor or tumor vasculature in the subject, The method of any of paragraphs 1-12, wherein the method is administered at a dose that causes one or more of:

  14. The method of any of paragraphs 1-13, wherein AHCM is losartan and is administered at 25-100 mg / day.

  15. The method of any of paragraphs 1-14, wherein the AHCM is losartan and is provided in a 12.5 mg, 25 mg, 50 mg, or 100 mg dosage form.

  16.AHCM is losartan, 0.25-17.5 mg / day, 0.5-15 mg / day, 1.3-12 mg / day, 1.5-12 mg / day, 2-12 mg / day, 2-10 mg / day, 2-5 mg / day 15. The method of any of paragraphs 1-15, administered at a non-hypertensive dose in the range of 2-3 mg / day, or 2 mg / day.

  17. AHCM is losartan, a dose that is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, more than 10 times the standard medical dose used for antihypertensive or anti-heart failure, The method of any of paragraphs 1-16, wherein the method is administered at or above a higher dose.

  18. The method of any of paragraphs 1-17, wherein AHCM is administered as a first course of treatment with a non-hypertensive dose of AHCM and then administered as a second course of treatment with AHCM over a standard antihypertensive dose.

  19.AHCM is administered as an entity with a hydrodynamic diameter greater than 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm , Any one of Items 1-18.

  20. The method of paragraph 19, wherein AHCM is administered as polymer nanoparticles or lipid nanoparticles.

  21. Cancer treatment is administered as an entity having a hydrodynamic diameter greater than 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm Item 21. The method according to any one of Items 1 to 20, which is a therapeutic agent for cancer.

  22. The method of paragraph 21, wherein the cancer therapeutic agent is administered as polymer nanoparticles or lipid nanoparticles.

  23. AHCM or cancer therapeutic agent each independently has a hydrodynamic diameter of 1 nm or less or 0.1 to 1.0 nm; a hydrodynamic diameter of 5 to 20 nm or 5 to 15 nm; or 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, Any of paragraphs 1-22, provided as an entity having a size range (nm) of hydraulic diameter greater than 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm Method.

24.
(I) not having hypertension,
(Ii) not having been treated for hypertension at the start of AHCM treatment; or (iii) having one or more of having normal or low blood pressure. Either way.

  25. Any of paragraphs 1-24, wherein the subject has not been administered AHCM within 5 days, within 10 days, within 30 days, within 60 days, or within 100 days of the start of cancer diagnosis or AHCM medication Method.

  26. The method of any of paragraphs 1-25, wherein the subject is in need of cancer treatment or is being considered for cancer treatment.

  27. The method of any of paragraphs 1-26, comprising determining whether the subject has cancer and administering AHCM and a cancer treatment in response to the determination.

  28. The method of any of paragraphs 1-27, wherein the subject is at risk of having a solid fibrous tumor or has a solid fibrous tumor.

  29. The method of paragraph 28, wherein the subject has a preneoplastic condition or is predisposed to cancer.

  30. Cancer is pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, prostate cancer, cervical cancer, gastrointestinal cancer, stomach cancer, head and neck cancer, kidney cancer, or liver cancer, or metastasis thereof The method according to any one of Items 1 to 29, which is selected from one or more of the lesions.

  31. The method of any of paragraphs 1-30, wherein AHCM is administered prior to and / or in combination with cancer treatment.

  32. The method of any of paragraphs 1-31, wherein the cancer treatment is selected from one or more of anti-cancer agents, surgery, and / or radiation.

  33. AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier , Any one of Items 1-32.

  34. The method of any of paragraphs 1-33, wherein the AHCM is maintained for a preselected portion of the time that the subject receives cancer treatment.

  35. The method of any of paragraphs 1-34, wherein the AHCM is maintained for the entire period during which the cancer treatment is administered.

  36. AHCM is at least 1 hour, 5 hours, 10 hours, or 24 hours; at least 2 days, 5 days, 10 days, or 14 days; at least 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks; Any of paragraphs 1-35, administered continuously over a period of at least 2 months, 3 months, 4 months, 5 months, or 6 months; or at least 1 year, 2 years, 3 years, 4 years, or 5 years That way.

  37. The method of any of paragraphs 1-36, wherein AHCM is administered as a sustained release formulation.

  38. The method of any of paragraphs 1-37, wherein the AHCM is formulated for oral delivery, subcutaneous delivery, or intravenous continuous delivery.

  39. The method of any of paragraphs 1-38, wherein AHCM is administered via a subcutaneous pump, implant, or depot.

40. Cancer treatment
(I) a viral cancer therapeutic agent, an anticancer therapeutic agent lipid nanoparticle, an anticancer therapeutic agent polymer nanoparticle, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a cancer therapeutic agent selected from a chemotherapeutic agent,
(Ii) radiation,
The method according to any one of Items 1 to 39, which is selected from one or more of (iii) surgery, or (iv) any combination of (i) to (iii).

  41. The method of paragraph 40, wherein the lipid nanoparticles are selected from PEGylated liposomal doxorubicin (DOXIL®) or liposomal paclitaxel (eg, Abraxane®).

  42. The method of paragraph 40, wherein the chemotherapeutic agent is selected from gemcitabine, cisplatin, epirubicin, 5-fluorouracil, paclitaxel, oxaliplatin, or leucovorin.

  43. The method of paragraph 40, wherein the antibody to the cancer target is selected from antibodies to HER-2 / neu, HER3, VEGF, or EGFR.

  44. A tyrosine kinase inhibitor selected from sunitinib, erlotinib, gefitinib, sorafenib, icotinib, lapatinib, neratinib, vandetanib, BIBW 2992, or XL-647, or cetuximab, panitumumab, sartumuzumab, 40. The method according to any one of Items 1 to 39, which is an anti-EGFR antibody selected from.

  45. The method of paragraph 40, wherein the chemotherapeutic agent is a cytotoxic agent or a cell division inhibitor.

  46. The method of paragraph 40 or 45, wherein the chemotherapeutic agent is selected from a microtubule inhibitor, a topoisomerase inhibitor, a taxane, an antimetabolite, a mitosis inhibitor, an alkylating agent, or an intercalating agent.

  47. The method according to any of paragraphs 1 to 46, wherein the cancer treatment is selected from one or more of an anti-angiogenic agent, a vascular targeting agent, or a vascular disrupting agent.

  48. AHCM or cancer treatment is administered to a subject by systemic administration selected from oral, parenteral, subcutaneous, intravenous, rectal, intramuscular, intraperitoneal, intranasal, transdermal, or by inhalation or intracavitary infusion. 48. The method according to any one of Items 1 to 47.

49. Tumor size;
The level or signaling of one or more of transforming growth factor β1 (TGFβ1), connective tissue growth factor (CTGF), or thrombospondin-1 (TSP-1);
Tumor collagen I level;
Fiber content;
Interstitial pressure;
A plasma or serum biomarker selected from collagen I, collagen III, collagen IV, TGFβ1, CTGF, or TSP-1;
The level of one or more cancer markers;
Emergence of new lesions, metabolism, rate of hypoxia generation;
The appearance of new disease-related symptoms;
Size of tissue mass;
Amount of disease-related pain;
Further monitoring the subject for one or more changes of histological analysis, leaflet pattern, and / or presence or absence of mitotic cells; or tumor invasiveness, primary tumor angiogenesis, or metastatic spread 49. The method according to any one of Items 1 to 48.

  49. A pharmaceutical composition comprising nanoparticles comprising AHCM.

  50. A pharmaceutical composition comprising nanoparticles comprising AHCM and a cancer therapeutic agent.

  51. The pharmaceutical agent of paragraph 50, wherein the cancer therapeutic agent is selected from a viral cancer therapeutic agent, an anticancer agent lipid nanoparticle, an anticancer agent polymer nanoparticle, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a chemotherapeutic agent. Composition.

  52. The pharmaceutical composition according to any one of Items 49 to 51, wherein the nanoparticles are polymer nanoparticles or lipid nanoparticles.

  53. The pharmaceutical composition of any of paragraphs 49-51, wherein AHCM is formulated in a dosage form according to the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure.

  54.0.01 times, 0.02 times, 0.03 times, 0.04 times, 0.05 times, 0.06 times, 0.07 times, 0.08 times, 0.09 times, 0.1 times the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure 52. The pharmaceutical composition of any of paragraphs 49-51, formulated in a dosage form smaller than fold, 0.15 fold, 0.16 fold, 0.2 fold, 0.3 fold, 0.4 fold, 0.5 fold, 0.6 fold, 0.7 fold.

  55. AHCM is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times greater than or equal to the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure Item 52. The pharmaceutical composition according to any one of Items 49 to 51, which is formulated in the above dosage form.

  56.0.01 times, 0.02 times, 0.03 times, 0.04 times, 0.05 times, 0.06 times, 0.07 times, 0.08 times, 0.09 times, 0.1 times the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure AHCM dosage form formulated in dosage forms smaller than double, 0.15 times, 0.16 times, 0.2 times, 0.3 times, 0.4 times, 0.5 times, 0.6 times, 0.7 times.

  57. AHCM is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times greater than or equal to the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure AHCM dosage form formulated in the above dosage form.

58. A method of optimizing the access of an agent, eg, a diagnostic or imaging agent, to a cancer or optimizing delivery to a cancer,
Administering a hypotensive and / or collagen modifying agent (“AHCM”) to the subject; and optionally administering the agent to the subject;
(A) the diagnostic or imaging agent has a hydraulic diameter greater than 1 nm, 5 nm, or 20-150 nm;
(B) the agent is a radiation agent, NMRA agent, contrast agent; or (c) the subject is at least 2 days, 3 days, or 5 days, or 1 week, 2 weeks prior to administration of the agent. Or one or more of being treated with AHCM administration initiated prior to administration of the agent for 3 weeks, 4 weeks, 5 weeks, or longer.

59. contacting the cancer or cancer-related cells with the candidate agent;
A method or assay for identifying an antihypertensive and / or collagen modifying agent (AHCM) comprising detecting a change in a cancer cell in the presence or absence of said candidate agent, comprising:
A method or assay wherein the detected change comprises one or more of increased or decreased active TGFβ, TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen (eg, collagen 1) levels.

60. Candidate drugs are renin angiotensin aldosterone antagonist (“RAAS antagonist”), angiotensin converting enzyme (ACE) inhibitor, angiotensin II receptor blocker (AT ₁ blocker), thrombospondin 1 (TSP-1) 60. The method or assay of paragraph 59, selected from one or more of an inhibitor, a transforming growth factor β1 (TGF-β1) inhibitor, or a connective tissue growth factor (CTGF) inhibitor.

  61. The method or assay of paragraph 59 or 60, wherein the candidate agent decreases one or more of active TGFβ, TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen levels.

  62. The method or assay of any of paragraphs 59-61, further comprising: comparing the treated method or assay to a reference value; and comparing the difference between the treated value and the reference value. .

  63. The method or assay of any of paragraphs 59-62, performed in vitro, in vivo, or a combination of both.

  64. Evaluate candidate drugs in vitro by adding candidate drugs to culture medium and analyzing conditioned media for increased or decreased active TGFβ, TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen levels 64. The method or assay of any of paragraphs 59-63, comprising a step.

  65. administering a candidate agent to an animal tumor model; and analyzing the subject for an increase or decrease in active TGFβ, TGFβ1 level, connective tissue growth factor (CTGF) level, or collagen level. Any of the methods or assays.

  66. Antihypertensive and / or collagen modifier (AHCM) compositions, or hypotensive and / or collagen modified alone or in combination with a cancer therapeutic, for use in the treatment of cancer alone or in combination with a cancer therapeutic Use of an agent (AHCM) to treat cancer.

  67. A therapeutic kit comprising an antihypertensive and / or collagen modifying agent (AHCM), alone or in combination with a cancer therapeutic agent, and instructions for the treatment of cancer.

  68. A diagnostic kit comprising a hypotensive and / or collagen modifying agent (AHCM), alone or in combination with an imaging agent, and instructions for the diagnosis of cancer.

69. selecting a subject as in need of receiving AHCM based on the need for improved delivery or efficacy of cancer treatment; and (a) administering AHCM to said subject, or (b) cancer A method of selecting a subject to receive an antihypertensive and / or collagen modifying agent (AHCM) comprising either or both of administering a treatment comprising:
The method wherein the AHCM is administered at a dosage sufficient to improve the delivery or efficacy of the cancer treatment.

  Having generally described the invention, the invention will be more readily understood by reference to the following examples. The examples are included solely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

  In Examples 1-6 below, we have found that AHCM agents, such as losartan, an angiotensin II receptor antagonist that is generally clinically approved for the treatment of hypertension, have anti-fibrotic effects, for example. It was assessed whether the penetration and efficacy of nanopharmaceuticals could be enhanced. Nanotherapeutics have shown new expectations for cancer treatment, but their clinical efficacy is modest (Jain RK, et al. (2010) Nat Rev Clin Oncol 7: 653-664; Davis ME, et al (2008) Nat Rev Drug Discov 7: 771-782; Peer D, et al. (2007) Nat Nanotechnol 2: 751-760; and Torchilin VP (2005) Nat Rev Drug Discov 4: 145-160). Without wishing to be bound by theory, a dense collagen network in a tumor can generally reduce the penetration and efficacy of nanotherapeutics. This is partly because small fiber spacings in the interstitium impede penetration, especially in fibrotic tumors that slow the movement of particles larger than 10 nanometers (Netti PA, et al. (2000 ) Cancer Res 60: 2497-2503; Pluen A, et al. (2001) Proc Natl Acad Sci USA 98: 4628-4633; Ramanujan S, et al. (2002) Biophys J 83: 1650-1660 and Brown E, et al. (2003) Nat Med 9: 796-800). The FDA-approved PEGylated liposomal doxorubicin (DOXIL®) and oncolytic virus currently undergoing multiple clinical trials, its size (approximately 100 nm) hinders intratumoral dispersion and therapeutic efficacy (Nemunaitis J, et al. (2001) J Clin Oncol 19: 289-298). Matrix modifiers such as bacterial collagenase, relaxin, and matrix metalloproteinases 1 and 8 have been used to modify collagen or proteoglycan networks in tumors, and the efficacy of intratumoral (it) injected tumor regression virus (Brown E, et al. (2003) Nat Med 9: 796-800; McKee TD, et al. (2006) Cancer Res 66: 2509-2513; Mok W, et al. (2007) Cancer Res 67 Ganesh S, et al. (2007) Cancer Res 67: 4399-4407; and Kim JH, et al. (2006) J Natl Cancer Inst 98: 1482-1493). Furthermore, relaxin can improve transport through the tumor matrix, but cannot facilitate the delivery of low molecular weight drugs (US Pat. No. 6,719,977). However, these agents may produce normal tissue toxicity (eg, bacterial collagenase) or increase the risk of tumor progression (eg, relaxin, matrix metalloproteinase).

  Losartan (Johnston CI (1995) Lancet 346: 1403-1407) approved to manage hypertension in patients does not have many of these safety risks. In addition to its antihypertensive properties, losartan is also an antifibrotic agent that has been shown to reduce the prevalence of heart and kidney fibrosis (Habashi JP, et al. (2006) Science 312: 117). -121; and Cohn RD, et al. (2007) Nat Med 13: 204-210). Losartan's antifibrotic effect is partly due to active transforming proliferation through downregulation mediated by angiotensin II type I receptor (AGTR1) of TGFβ1 activators such as thrombospondin 1 (TSP-1) Caused by suppression of factor β1 (TGFβ1) levels (Habashi JP, et al. (2006) Science 312: 117-121; Cohn RD, et al. (2007) Nat Med 13: 204-210; Lavoie P, et al (2005) J Hypertens 23: 1895-1903; Chamberlain JS (2007) Nat Med 13: 125-126; and Dietz HC (2010) J Clin Invest 120: 403-407).

  As demonstrated below, AHCM agents, such as losartan, inhibited collagen I production by cancer-associated fibroblasts (CAF) isolated from breast cancer biopsies. Furthermore, AHCM agents, such as losartan, resulted in a dose-dependent decrease in matrix collagen in a fibrogenic model of human breast, pancreas, and skin tumors in mice. In addition, AHCM agents such as losartan improved the dispersion and therapeutic efficacy of intratumorally injected oncolytic herpes simplex virus (HSV). In addition, AHCM agents such as losartan also enhanced the efficacy of intravenously injected PEGylated liposomal doxorubicin (DOXIL®). Thus, administration of an AHCM agent, eg, losartan, in combination with a cancer therapeutic agent (eg, a cancer nanotherapeutic agent) can enhance the efficacy of the nanotherapeutic agent in patients with fibrogenic tumors.

  Using doses with minimal effect on mean arterial blood pressure (MABP), we have developed four tumor models: spontaneous mouse breast cancer (FVB MMTV PyVT), orthotopic pancreatic adenocarcinoma ( L3.6pl), and in fibrosarcomas implanted subcutaneously (HSTS26T) and melanoma (Mu89)), AHCM agents, such as losartan, have shown below to reduce collagen I levels. In addition, the inventors have shown below that AHCM agents, such as losartan, can also improve intratumoral penetration of nanoparticles injected intratumorally (i.t.) or intravenously (i.v.).

  In addition, the inventors have described it administration (methods of administration to patients widely used for gene therapy) (Hu JC, et al. (2006) Clin Cancer Res 12: 6737-6747; Senzer NN, et al. (2009) J Clin Oncol 27: 5763-5771; Breitbach CJ, et al. (2010) Cytokine Growth Factor Rev 21: 85-89) Dispersion and efficacy of tumor regression HSV and iv administered DOXIL ( It was assessed how AHCM agents, such as losartan, affect the efficacy of the registered trademark. As shown below, AHCM agents, such as losartan, improved the efficacy of both i.t. injected tumor regression HSV and i.v. administered DOXIL®. Results from intratumoral (i.t.) experiments indicate that AHCM agents, such as losartan, can enhance nanoparticle penetration in the interstitial space by improving interstitial transport. Furthermore, findings from the intravenous (iv) study show that AHCM agents (eg, losartan) show the efficacy of systemically administered nanotherapeutic drugs against fibrotic solid tumors, highly fibrotic solids such as pancreatic adenocarcinoma. Even tumors can be improved. Thus, AHCM agents such as losartan, an antihypertensive drug approved by the FDA, can be used to improve the efficacy of various nanotherapeutic agents in multiple tumor types.

Example 1: Losartan inhibits collagen I synthesis by cancer-associated fibroblasts (CAF) The expression and activation of TGFβ1 by breast CAF and the effect of losartan on collagen I production were investigated (FIG. 1). Losartan reduces TGFβ1 activation and collagen I production in cancer-associated fibroblasts in vitro. Cells were treated with 10 μmol / L losartan for 24 hours. Losartan reduced active TGFβ1 levels by 90%, but total TGFβ1 levels were not affected. There was a corresponding 27% decrease in collagen I levels. The decrease in active TGFβ1 and collagen I was statistically significant (Student t test p <0.05). Since collagen in tumors is mainly produced by CAF, the effect of losartan on collagen content in tumors was investigated.

Example 2: Losartan reduces collagen I in tumors in a dose-dependent manner To determine the dose response of losartan to intratumoral collagen levels, 10 mg / kg / day, 20 mg / kg / day, and 60 mg / kg / day Losartan intraperitoneal (ip) injection, second harmonic generation (SHG) imaging of fibrous collagen in HSTS26T tumor in dorsal skinfold chamber (Figures 2A-2B) and collagen I immunostaining of tumor sections (Figures 3A-3D) Carried out. SHG signal strength can include signals contributed by collagen I and other fibrogenic collagens (eg, collagen III or V), but collagen I is generally the predominant collagen type in most soft tissues. Yes (Gelse K, et al. (2003) Adv Drug Deliv Rev 55: 1531-1546) and therefore contribute as the main source of the SHG signal. Furthermore, in human pancreatic tumors, collagen I is the main fibrous collagen, and the level of collagen V is significantly lower (Mollenhauer J, et al. (1987) Pancreas 2: 14-24). Losartan doses at 20 mg / kg / day and 60 mg / kg / day significantly reduced intratumoral SHG signal intensity, while the lowest dose of 10 mg / kg / day had a significant effect on SHG signal intensity None (Figures 2A and 2B). Injection of 60 mg / kg / day and 20 mg / kg / day of losartan also significantly reduced collagen I immunostaining in HSTS26T tumors by 65% and 42%, respectively (FIG. 3). Treatment with the 60 mg / kg / day dose resulted in the highest degree of decrease in collagen I and reduced mean arterial blood pressure (MABP) by 35 mmHg (p <0.04; FIG. 4). In the following examples, a 20 mg / kg / day dose was used (but it is in no way limiting the use of other doses in the methods described herein). After 2 weeks of losartan treatment, a dose of 20 mg / kg / day reduced MABP by 10 mmHg (Figure 4) and therefore maintained MABP within the normal range (70-95 mmHg) for SCID mice (Kristjansen PE et al. (1993) Cancer Res 53: 4764-4766). It also had no detectable effect on mouse body weight (mean 26 ± 1 g treatment vs. 26 ± 1 g control). The 20 mg / kg / day dose was determined for 47% (p <0.05) and 50% (p, respectively) in four tumor types (FVB MMTV PyVT, L3.6pl, HSTS26T, and Mu89). <0.03), 44% (p <0.04), and 20% (p <0.02) (Figures 5A-5D).

Example 3: Losartan reduces TSP-1 expression in tumors
TSP-1 is an important regulator of TGFβ1 activation, and losartan has been reported to reduce TSP-1 expression and TGFβ1 activation in a mouse model of Marfan syndrome and muscular dystrophy (Dietz HC (2010 ) J Clin Invest 120: 403-407). As shown herein, measurement of protein levels in homogenized HSTS26T tumors showed that losartan did not affect total TGFβ1 levels, but significantly increased levels of TSP-1, active TGFβ1, and collagen I. It was shown to decrease (Figure 6). Losartan also reduced TSP-1 immunostaining in HSTS26T (73%, p <0.04) and Mu89 (24%, p <0.03) (FIGS. 7A-7B). In both Mu89 and HSTS26T tumors, the immunostaining patterns for TSP-1 (FIGS. 7A-7B) and collagen I (FIGS. 5C-5D) matched closely. Although we detected high levels of TSP-1 and collagen I in the tumor periphery, losartan induced a clear decrease in TSP-1 and collagen I levels in the tumor center (FIGS. 5C, 7A). ). These data indicate that the decrease in collagen I levels is due in part to the decrease in TGFβ1 activation due to the loss induced by losartan in TSP-1 expression.

Example 4: Losartan Improves Intratumoral Dispersion of Nanoparticles and Nanotherapeutic Agents Based on previous studies on tumor stromal matrices (Pluen A, et al. (2001) Proc Natl Acad Sci USA 98: 4628- 4633 and Brown E, et al. (2003) Nat Med 9: 796-800), we will determine if a decrease in collagen content by losartan can improve intratumoral dispersion of nanoparticles. I made an effort. Therefore, we measured the intratumoral dispersion of fluorescent polystyrene nanoparticles (100 nm diameter) after it injection or iv injection in three different tumor types (HSTS26T, Mu89, and L3.6pl). In mice injected with nanoparticles, losartan improved nanoparticle accumulation and penetration at the tumor center (FIG. 8A; HSTS26T p <0.001, Mu89 p <0.001). Conversely, there was little or no nanoparticle accumulation in the center of the control tumor. In control tumors, the majority of injected nanoparticles were found around the tumor periphery and around the needle insertion point (FIG. 8A). We also determined the effect of losartan on intratumoral dispersion of tumor regression HSV. In both HSTS26T and Mu89, losartan significantly increased the intratumoral spread of HSV injected intratumorally (FIG. 8B). These findings indicate that losartan increases the dispersion of large nanoparticles, and we found that in HSTS26T, losartan increased IgG interstitial diffusion (Figure 9) and It was also determined to increase the calculated average stromal matrix pore size from 9.91 ± 0.43 nm to 11.78 ± 0.41 nm (Nugent LJ, et al. (1984) Cancer Res 44: 238-244).

  The inventors then assessed the effect of losartan on vascular perfusion and intratumoral dispersion of i.v. injected nanoparticles in mice with orthotopic pancreatic tumors (L3.6pl). In tumors treated with losartan, the accumulation and penetration of beads away from blood vessels within the tumor was significantly higher (FIGS. 8C and 10). These results indicate that losartan improves the transport and dispersion of both i.t. injected nanoparticles and / or i.v. injected nanoparticles.

Example 5: Losartan Improves Potency of DOXIL® and Tumor Regression HSV Next, we have determined that losartan is a tumor regression HSV injected with it and a potency of DOXIL® injected with iv. It was decided whether or not it could be improved. The effect of losartan combined with it injection of HSV was determined in HSTS26T and Mu89 tumors. Administration of losartan alone did not affect the rate of tumor growth (FIGS. 11A and 11B). However, when animals were treated with losartan for 2 weeks prior to it injection of HSV, losartan significantly delayed the increase in both Mu89 and HSTS26T tumors (FIGS. 11A and 11B). In 50% of mice treated with losartan and HSV, HSTS26T tumor volume remained stable for up to 9 weeks. For Mu89 tumors, mice treated with losartan and HSV had a delay in tumor growth. However, the growth delay in Mu89 tumors was only transient, with all tumors 3 times larger than the starting treatment size 4 weeks after virus injection.

  To determine whether losartan could increase the efficacy of i.v. injected nanotherapeutics, mice with orthotopic pancreatic tumors (L3.6pl) were treated with DOXIL® and losartan. After 4 weeks of tumor implantation, 2 weeks after the start of losartan treatment (20 mg / kg / day), we have developed a non-anti-tumor dose (ie, a dose that is not effective for the treatment of cancer, eg, tumor growth Mice were treated with DOXIL® (4 mg / kg, iv) at a dose and / or a dose not effective to inhibit or prevent progression. After 7 days, losartan or DOXIL alone did not affect mean tumor weight (FIG. 11C). However, tumors were significantly smaller in mice treated with losartan and DOXIL® than in mice that received DOXIL® alone (p <0.001) (FIGS. 11C and 11D).

Example 6: Pattern of collagen dispersion modulates the efficacy of losartan To investigate the difference between HSTS26T and Mu89 in response to the losartan-HSV combination treatment, we have 21 days after it injection of HSV HSV infection and necrosis patterns were determined. Figures 12A and 12B show significant differences in collagen structure in Mu89 tumor (Figure 12A) and HSTS26T tumor (Figure 12B), respectively. While not wishing to be bound by theory, these differences in collagen structure have altered virus propagation in these tumor types. In Mu89 tumors, the collagen fiber network was well organized and formed a finger-like projection into the tumor (FIGS. 12A and 13A). These protrusions divide the tumor into separate compartments that cannot be traversed by HSV particles, thus limiting viral infection and consequent necrosis (FIG. 14A). Losartan treatment, to some extent, destroyed collagen processes but did not completely eliminate them (FIG. 12A). As a result, in Mu89 tumors treated with losartan, there was some degree of crossing of virus particles between compartments. Varying the concentration of losartan and / or HSV modulates the amount or extent of collagen processes destroyed (eg, partial or complete destruction), and thus, in the distribution of HSV particles within the tumor Can influence. In contrast, in HSTS26T tumors, the dense collagen network was more diffuse, less fibrous, and less compartmentalized (FIGS. 12B and 13B). In this tumor, the dense collagen network appeared to slow down virus propagation but did not completely interfere, resulting in increased virus propagation and a more diffuse pattern of necrosis (FIG. 14A).

Discussion The renin angiotensin aldosterone system (RAAS) has been reported to play a role in the regulation and production of extracellular matrix components (Cook KL, et al. (2010) Cancer Res 70: 8319-8328; Rodriguez-Vita J , et al. (2005) Circulation 111: 2509-2517; and Wolf G (2006) Kidney Int 70: 1914-1919). Angiotensin II has been reported to stimulate collagen production through both TGFβ1-dependent and independent pathways (Yang F, et al. (2009) Hypertension 54: 877-884). Losartan and other RAAS inhibitors reduce the levels of collagen I and III and basement membrane collagen IV in various experimental models of fibrosis (Toblli JE, et al. (2002) J Urol 168: 1550-1555 and Boffa JJ, et al. (2003) J Am Soc Nephrol 14: 1132-1144) has been reported to reverse kidney and heart fibrosis in hypertensive patients (Lim DS, et al. (2001) Circulation). 103: 789-791 and Khalil A, et al. (2000) J Urol 164: 186-191). Using four different tumor types, we have demonstrated for the first time that losartan also inhibits collagen I production in tumors.

  Other matrix modifiers such as bacterial collagenase, relaxin, and matrix metalloproteinases 1 and 8 have been reported to modify the network of collagen or proteoglycans in tumors, and have shown the efficacy of intratumorally injected tumor regression virus. Improved (Brown E, et al. (2003) Nat Med 9: 796-800; McKee TD, et al. (2006) Cancer Res 66: 2509-2513; Mok W, et al. (2007) Cancer Res 67: 10664-10668; Ganesh S, et al. (2007) Cancer Res 67: 4399-4407; and Kim JH, et al. (2006) J Natl Cancer Inst 98: 1482-1493). However, these agents may produce normal tissue toxicity (eg, bacterial collagenase) or increase the risk of tumor progression (eg, relaxin, matrix metalloproteinase). In contrast, losartan (Johnston CI (1995) Lancet 346: 1403-1407) has limited side effects. Losartan has been reported to reduce the prevalence of metastases in several tumor types (Arafat HA, et al. (2007) J Am Coll Surg 204: 996-1005).

  As shown in the above examples, AHCM agents, such as losartan, can reduce collagen content and then improve interstitial transport and intratumoral dispersion of nanoparticles and nanotherapeutics. The inventors have also discovered that the organization of the collagen fiber network can affect nanoparticle dispersion. This was striking because of the significant difference in the structural organization of fibrous collagen I between Mu89 and HSTS26T. In Mu89 tumors, a thick bundle of fibrous collagen I surrounds the periphery of the tumor, forming a finger-like projection that subdivides the tumor mass into an isolated compartment and isolates the viral infection at the injection site / isolation Restricted to closed compartments (Figures 12A and 13A). In contrast, HSTS26T tumors have a reticulated collagen structure that prevents viral spread but does not restrict viral particles to the injection site (FIGS. 12B and 13B). The slower growth rate of HSTS26T tumors than Mu89 tumors may also explain, in part, the enhanced efficacy of losartan combined with HSV in HSTS26T tumors. Thus, not only the collagen content, but also the organization of the collagen network plays an important role in limiting the penetration of large therapeutic agents in the tumor. Depending on the content and / or organization of the collagen network in certain tumors, the dose, mode of administration, and / or frequency of AHCM agents (eg, losartan) and / or cancer therapeutic agents (eg, HSV) may vary. Can be adjusted.

  Pancreatic cancer patients treated with cytotoxic agents have a very high frequency of recurrence and a 5-year survival rate of less than 5% (Li J, et al. (2010) AAPS J 12: 223-232). Insufficient vascular supply and increased fiber content of pancreatic tumors are most likely to play a significant role in limiting the delivery and efficacy of cytotoxic drugs (Olive KP, et al. (2009) Science 324: 1457 -1461). We show that losartan increases both intratumoral dispersion and extravascular penetration distance of i.v. injected nanoparticles in a mouse orthotopic model of human pancreatic cancer (L3.6pl). Increased dispersion of nanoparticles and extravasation indicate that losartan not only improves interstitial transport, but also transvascular transport, as demonstrated by i.t. injection of nanoparticles and virus. When used alone, losartan did not affect the growth of pancreatic tumors or the body weight of treated mice. However, losartan combined with DOXIL® reduced tumor size by 50% compared to DOXIL® treatment alone. These findings indicate that losartan increased tumor penetration and dispersion of i.v. injected DOXIL® and enhanced efficacy in orthotopic pancreatic cancer in mice.

  Losartan's effect is not limited to interstitial space. Modifications to the RAAS system can either inhibit angiogenesis (Fujita M, et al. (2005) Carcinogenesis 26: 271-279) or alter tumor blood flow (Jain R, et al. (1984). ) IEEE Trans Son Ultrason 31: 504-526 and Zlotecki RA, et al. (1993) Cancer Res 53: 2466-2468). Inhibition of AGTR1 by losartan can also reduce the production of VEGF by cancer cells and the expression of VEGFR1 in endothelial cells and inhibit tumor angiogenesis and growth (Otake AH, et al. (2010) Cancer Chemother Pharmacol 66 : 79-87 and Noguchi R, et al. (2009) Oncol Rep 22: 355-360). As shown herein, losartan did not affect tumor growth or vascular density in HSTS26T tumors. Losartan can also reduce the growth of tumor cells expressing AGTR1 (Rhodes DR, et al. (2009) Proc Natl Acad Sci USA 106: 10284-10289). We found no reduction in cancer cell proliferation (FIG. 15) or tumor size (FIG. 16) in human melanoma Mu89 expressing AGTR1. Differences between those studies and other past studies may be due to differences in dosage. For example, in previous studies the dose of losartan was up to 15 times higher than that used in our study (Otake AH, et al. (2010) Cancer Chemother Pharmacol 66: 79-87). We have found that low-dose losartan, which alone is ineffective for the treatment of cancer, can be used for cancer treatment (even at non-therapeutic levels) to improve the efficacy of cancer therapy or anticancer agents. Is shown herein. Furthermore, low dose losartan allows for a more clinically translatable protocol and avoids hypotensive complications.

  Patients receiving RAAS antagonists have a reduced prevalence of breast and lung cancer (Lever AF, et al. (1998) Lancet 352: 179-184). Different mechanisms have been reported to discuss the antitumor properties of RAAS antagonists when used at high concentrations (Ager EI, et al. (2008) Carcinogenesis 29: 1675-1684; Lindberg H, et al. 2004) Acta Oncol 43: 142-152; Miyajima A, et al. (2002) Cancer Res 62: 4176-4179, and Rosenthal T, et al. (2009) J Hum Hypertens 23: 623-635). AGTR1 signaling has been reported to increase the proliferation of stromal and tumor cells and the transcription of inflammatory cytokines and chemokines that promote cancer cell migration and dissemination (Deshayes F, Nahmias C (2005) Endocrinol Metab 16: 293-299). Reduction of active TGFβ1 levels by RAAS antagonists administered at high concentrations has been reported to reduce metastasis (Jakowlew SB (2006) Cancer Metastasis Rev 25: 435-457). Thus, in addition to improving the delivery of anti-tumor agents, losartan can also inhibit tumor progression and metastasis. Merely by way of example, losartan administered at a low dose (eg, a dose that is not effective to reduce or prevent metastasis when administered alone) (eg, for treatment and / or prevention of metastasis). Can be used to inhibit tumor progression and metastasis with an antimetastatic agent (less than the dose typically administered alone).

  In order to use losartan as an adjunct in the treatment of cancer patients, it is important to consider medication and treatment schedules along with possible side effects. Results from dose- and time-dependent studies presented herein indicate losartan administration for a minimum of 2 weeks prior to anti-tumor treatment. It may be advisable to start Losartan treatment two weeks before the anti-tumor treatment schedule and continue it throughout the anti-tumor treatment schedule to obtain maximum effect in the patient. Long-term losartan treatment in hypertensive patients has been shown to have limited manageable side effects, and many anti-tumor agents (eg, anti-VEGF drugs) have been shown to increase blood pressure (Ager EI, et al. (2008) Carcinogenesis 29: 1675-1684), long-term co-treatment with losartan may be beneficial for cancer patients. In some embodiments, the patient is commonly used for the treatment of patients with Marfan syndrome (Brooke BS, et al. (2008) N Engl J Med 358: 2787-2795), 2 mg / kg / day It can be treated with a dose of losartan.

  Losartan and ARB have limited side effects, but losartan treatment is not recommended for patients with known kidney disease. Losartan may induce renal failure or congestive heart failure in patients with microvascular or macrovascular renal disease (Sica DA, et al (2005) Clin Pharmacokinet 44: 797-814). Hyperkalemia may occur in patients with poor renal function or in patients who are simultaneously receiving potassium supplements or potassium-sparing diuretics. Finally, in patients treated with losartan, angioedema caused by high levels of circulating angiotensin II may occur (Sica DA, et al (2005) Clin Pharmacokinet 44: 797-814).

  Tumor drug resistance is generally thought to occur at many levels, including increased drug efflux, drug inactivation, avoidance of apoptosis, and alteration of the target pathway (Longley DB, et al. (2005) J Pathol 205 : 275-292). Because losartan is not an anti-tumor agent, tumor resistance to losartan therapy after long-term treatment may be due to other mechanisms. Since TGFβ1 activation is induced by different agents such as MMPs and integrins in addition to TSP-1, tumor resistance to losartan may be due to changes in TGFβ1 activation and signaling. However, long-term losartan treatment after myocardial infarction has been reported not to be associated with reduced antifibrotic properties (Schieffer B, et al. (1994) Circulation 89: 2273-2282).

  As shown in Examples 1-6, we found that losartan reduced the matrix collagen content in the tumor, and both of it and iv delivered nanoparticles (DOXIL®, HSV) Shows improved penetration and therapeutic efficacy. Losartan also exhibits vasoactive and anti-metastatic properties that may increase its clinical application. Furthermore, since losartan has already been approved for clinical use, it represents a safe and effective aid for improving the efficacy of nanotherapeutics in cancer patients.

Exemplary Experimental Protocols for Examples 1-6 Exemplary Materials and Methods A more detailed description of the technique is presented in the Additional Materials and Methods section below.

  Briefly, CAF isolated from human breast cancer biopsy material was treated with losartan for 24 hours before measuring collagen and cytokine levels. The protein assay was performed using a commercially available ELISA kit. All animal experiments were performed with the approval of the Institutional Animal Care and Use Committee. Losartan was administered i.p. at concentrations of 10 mg / kg / day, 20 mg / kg / day, or 60 mg / kg / day for up to 2 weeks. After 2 weeks of losartan treatment, mice were treated with HSV (i.t.) and DOXIL® (intravenous via the tail vein). Resected tumors were snap frozen for biochemical analysis or fixed with paraformaldehyde and embedded with paraffin or optimal cutting temperature compound (OCT) for immunohistochemistry.

Additional materials and methods:
Cell culture CAF was isolated from human breast cancer biopsies using art-recognized protocols such as those described in Orimo A, et al. (2005) Cell 121: 335-348. . CAF was seeded in 24-well plates at a concentration of 500K cells / well. Cells were allowed to attach to the plate for 24 hours before 10 μmol / l losartan was added for 24 hours (Schuttert JB, et al. (2003) Pflugers Arch 446: 387-393). Treatment was performed in low serum to reduce background collagen levels. At the end of the 24 hour treatment period, conditioned media was collected and analyzed for collagen levels.

Protein Assay Collagen I measurements were performed using a type I C-terminal collagen propeptide enzyme-linked immunosorbent assay (ELISA) kit (Quidel, San Diego, CA) and a Sircol soluble collagen assay (Biocolor Ltd., United Kingdom) . The TGFβ1 assay was performed using a human TGFβ1 ELISA kit (R & D Systems, Minneapolis, Minn.). The assay measures only free mature TGFβ1. To measure the total level of TGFβ1, the latent form of TGFβ1 was activated with 1N HCl. The TSP-1 assay was performed using a human TSP-1 ELISA kit (R & D Systems, Minneapolis, Minn.).

Mice and tumor models All experiments were performed with the approval of the Institutional Animal Care and Use Committee. Human soft tissue sarcoma (HSTS26T) and human melanoma (Mu89) tumors were augmented subcutaneously in the legs and dorsal skinfold chambers of severe combined immunodeficiency (SCID) mice (Leunig M, et al. (1992) Cancer) Res 52: 6553-6560). Human pancreatic adenocarcinoma cells (L3.6PL) were augmented in the pancreas of SCID mice. L3.6PL tumors were induced by subcapsular injection of 1 million cells into the pancreatic tail. Tumor size was monitored in spontaneous FVB / N-Tg (MMTV-PyVT) 634MU1 / J mice and tumors were selected for treatment when they reached a size of 4-6 mm in diameter (Guy CT et al. 1992) Mol Cell Biol 12: 954-961).

Preparation and treatment of losartan
Cozaar (Losartan potassium) tablets were crushed using a mortar and pestle. The powder was then dissolved in water to obtain a concentration of 2.5 mg / ml. The solution was then filtered and stored in a sterile container. Losartan at a concentration of 10 mg / kg / day, 20 mg / kg / day, or 60 mg / kg / day was administered by ip injection daily for up to 2 weeks (Melo LG, et al. (1999) Am J Physiol 277: R624-R630).

Tissue collection, embedding, and staining Tumors for immunostaining analysis and quantification were collected from mice, fixed with 4% paraformaldehyde, paraffin or optimal cutting temperature compound (OCT) (Sakura Finetek Torrance, CA) Embedded in. OCT-embedded tumors were immersed in sucrose solution for 24 hours before embedding and freezing.

Immunostaining of collagen I and TSP-1 in frozen sections Frozen sections were cut into 10 μm sections for immunohistochemistry and imaging. Collagen I was detected using the LF-67 antibody (1: 100 dilution) according to the protocol previously described (Znati CA, et al. (2003) Clin Cancer Res 9: 5508-5513). TSP-1 was detected with a goat anti-human antibody (sc-12312, Santa Cruz Biotechnology Inc., Santa Cruz, Calif.) (1:50 dilution) that cross-reacts with mice. Images of 20x magnification were randomly obtained from each slide for analysis of collagen and TSP-1. Collagen and TSP-1 content was determined by measuring the number of pixels above a set threshold based on the average intensity value of the pixels from all analyzed slides. The background signal intensity was low and uniform for both collagen I and thrombospondin 1 immunostaining. We confirmed that the average signal intensity threshold provided an accurate representation of collagen and thrombospondin 1 immunostaining and did not contain background signal.

Second Harmonic Imaging of Collagen Fibers Second Harmonic Imaging (SHG) imaging was performed on dorsal chamber tumors with a custom-built multiphoton laser scanning microscope (Brown E, et al. (2003) Nat Med 9: 796-800) . Polarized light from a Ti: sapphire laser (Mai-Tai Broadband: Spectra-Physics, Mountain View, CA) was converted to circularly polarized light using a zero order quarter wave plate (Newport Corporation, Irvine, CA). An excitation wavelength of 810 nm and a SHG signal detected at 405 nm were used. SCID mice bearing HSTS26T tumors in dorsal chambers were treated with losartan (10 mg / kg / day, 20 mg / kg / day, or 60 mg / kg / day) or saline during the dose response experiment (15 days) Treated with either. To define 4 regions of interest in each mouse, vascular markers were used and periodically returned to the same region of SHG imaging. SHG images were analyzed with a custom-made Matlab (The MathWorks, Inc., Natick, MA) code. The fraction of the region of interest (ROI) that was SHG signal positive was normalized to the amount of SHG signal obtained on day 1 of the dose response study (prior to initiation of treatment with losartan or saline). .

Analysis of HSV infection and nanoparticle dispersion Intratumoral injection: Nanoparticles and tumor regression HSV were injected with a syringe pump (Harvard Apparatus Standard Pump 22, Holliston, Massachusetts) at a flow rate of 4 μl / min. We injected 10 μl of green fluorescent protein (GFP) expressing HSV (2.5 × 10 ⁵ tu), or 10 μl of fluorescent nanoparticles (100 μm diameter; 1 × 10 ¹³ nanoparticles / ml concentration). The injected tumor was excised 30 minutes after nanosphere injection and 24 hours after HSV infusion. The resected tumor was bisected at an angle perpendicular to the needle track, fixed with paraformaldehyde, and frozen with OCT. All tumor sections were obtained perpendicular to the needle track angle. The entire tumor section was imaged with a double confocal microscope (Olympus BX61WI) and the image was reconstructed as a mosaic. Nanosphere dispersion and GFP positive areas (HSV infected cells) correspond to the fraction of pixels brighter than the background signal.

Intravenous injection: A total volume of 10 μl at a concentration of 3.6 × 10 ¹³ nanoparticles / ml was injected via the tail vein. After 24 hours, 50 μl of FITC-lectin was injected to identify functional blood vessels. Five minutes after lectin injection, tumors were excised, fixed with paraformaldehyde, and embedded with OCT. The tumor was then sectioned before confocal imaging and analysis. The degree of nanosphere dispersion was determined by measuring the fraction of pixels brighter than the background signal. Nanosphere penetration was determined by contouring around the perfused blood vessel and recording the fraction of nanosphere positive pixels within each contour. The contour was expanded outward by 30 μm for each perfused blood vessel. Using a previously described algorithm (Tong RT, et al. (2004) Cancer Res 64: 3731-3736), we fit a plot of nanosphere fractions and distances from blood vessels to an exponential function. And the relative penetration depth of the nanospheres from each vessel was obtained.

式中、Dは腫瘍におけるプローブ分子についての拡散係数であり、D₀は水中の拡散係数であり、λはプローブの水力学的半径と孔の半径との比率である（Nugent LJ,et al.(1984)Microvasc Res 28:270-274）。 Diffusion measurements by recovery after fluorescence fading Mice with HSTS26T tumors implanted in dorsal skin fold chambers were treated with ip injections of losartan (40 mg / kg / day) for 1 week. Based on a protocol previously described (Chauhan VP, et al. (2009) Biophys J 97: 330-336), fluorescence post-fading recovery (FRAP) measurements were performed with a custom-made multiphoton microscope. IgG labeled fluorescein isothiocyanate (0.5 ml; 2 mg / ml) was injected it and used as a tracer. About 10 minutes after injection, diffusion was measured by multiphoton FRAP (MP-FRAP) and spatial Fourier analysis FRAP (SFA-FRAP). Matrix pore size was calculated using SFA-FRAP data using the following equation:

Where D is the diffusion coefficient for the probe molecule in the tumor, D ₀ is the diffusion coefficient in water, and λ is the ratio of the probe hydrodynamic radius to the pore radius (Nugent LJ, et al. (1984) Microvasc Res 28: 270-274).

Analysis of HSV infection, necrosis, and collagen structure
To determine the relationship between viral infection, necrosis and collagen structure 21 days after HSV injection, serial paraffin sections were either polyclonal HSV-1 antibody (DAKO, Glostrup Denmark) or collagen I antibody (LF-67). Stained by The slides were treated with 3% hydrogen peroxide for collagen I staining in paraffin sections, and then antigen activation with Target Retrieval Solution, pH 9 (DAKO, Carpinteria, CA). The slide was then treated with 0.05% trypsin and then primary collagen I antibody was applied at a dilution of 1: 500. Sections stained with collagen I or HSV were imaged by light microscopy.

、逆方向プライマー

を使用した。 PCR analysis
RNA was extracted using the RNeasy mini kit (Qiagen, Valencia, Calif.) And converted to cDNA using the RT ² first strand kit (SuperArray Biosciences Corporation, Frederick, MD). The quality and concentration of cDNA was measured with an ND-200 spectrophotometer (Nanodrop Technologies, Wilmington, DE). For PCR reactions, cDNA from all samples was normalized to 1 μg / μl. The reaction was performed with HotStarTaq Plus DNA Polymerase (Qiagen, Valencia, CA). For AGTR1 primer, we have forward primer

, Reverse primer

It was used.

Ki67 staining, imaging and analysis
Ki67 staining was performed on paraffin sections 21 days after HSV injection. The slide was subjected to microwave treatment with Target Retrieval Solution (DAKO, Carpinteria, CA), and then primary antibody detection was performed. The entire tumor section was imaged at 2x magnification and reconstructed as a mosaic. Twenty regions were randomly selected for each tumor. The fraction of Ki67 positive cells in each region was determined by manual counting.

DOXIL® treatment and delayed tumor growth Two weeks after implantation of orthotopic pancreatic L3.6PL tumors, mice were randomly selected for treatment with losartan or saline. After 2 weeks of losartan treatment (20 mg / kg / day), a non-anti-tumor dose of DOXIL® (4 mg / kg) was injected iv via the tail vein. One week after DOXIL injection, tumors were excised and measured.

Virus treatment and delayed tumor growth Scid mice bearing HSTS26T and MU89 tumors subcutaneously were randomly divided into control and losartan treatment groups. Each arm (control and treatment) was then divided into HSV and non-HSV treatment groups. Tumors that reached 60 mm ³ after 2 weeks were selected for itHSV injection. Tumors were treated with 10 μl it injection of either PBS or 2.5 × 10 ⁵ transducing units (tu) of tumor regression HSV MGH2 (a gift from E. Antonio Chiocca, Ohio State University, Columbus, OH). Twice it injection of tumor regression HSV was administered at 24 hour intervals. Injections were made using a Harvard Apparatus Standard Pump 22 infusion / aspiration syringe pump system (Holliston, Massachusetts) at a flow rate of 4 μl / min. Tumors were measured every 2-3 days. Tumor volume was estimated as follows: V = AB ^2/2. Where V is the tumor volume; A and B are the maximum and minimum diameters of the tumor as measured by calipers.

Statistics All animal experiments were performed with at least 6 mice in each treatment arm. Tumor growth delay studies in HSTS26T and MU89 tumors were performed with at least 8 mice in each group. Our rationale for the number of mice used showed that we needed at least 8 mice in each group to reach statistical significance (p <0.05). Based on power calculations from previous studies (McKee TD, et al. (2006) Cancer Res 66: 2509-2513 and Mok W, et al. (2007) Cancer Res 67: 10664-10668). All statistical analyzes including the two groups were performed using the Student t test. A p value of less than 0.05 was considered significant. For groups, Tukey's post-hoc test after one-way Anova test was used to determine statistical significance between groups. Statistical significance in the figure is identified by an asterisk (“ ^* ”).

Example 7: Angiotensin inhibition improves drug delivery by normalizing the tumor microenvironment Advances in biomedical research have led to several new systemically administered molecular therapeutics and nanotechnology in both preclinical and clinical environments This led to the introduction of therapeutic agents (Jones, D. (2007) Nat Rev Drug Discov 6,174-175; Moghimi, SM et al. (2005) Faseb J 19,311-330). Although these new agents act on unique targets that give greater specificity or improved pharmacodynamic properties to tumor cells, their effectiveness limits delivery due to the characteristics of the tumor microenvironment. (Jain, RK (1998) Nat Med 4,655-657; Sanhai, WR et al. (2008) Nat Nanotechnol 3,242-244). The mechanical force induced by the increase compresses and collapses the blood vessels, limiting tumor perfusion, and the abnormal vasculature results in heterogeneous drug extravasation. The third determinant of delivery, interstitial transport through tissues, is disturbed specifically by nanomedicine in tumors due to abnormally dense and refracted tumor stroma (Jain, RK & Stylianopoulos (2010) Nat Rev Clin Oncol. 139; Chauhan, VP et al. (2009) Biophysical journal 97, 330-336). These barriers include pancreatic cancer (Olive, KP et al. (2009) Science 324,1457-1461), colorectal cancer (Halvorsen, TB & Seim, E. (1989) J Clin Pathol 42,162-166), lung cancer and breast cancer ( Ronnov-Jessen, L. et al. (1996) Physiol Rev 76, 69-125), a drug that specifically affects the treatment for patients with fibrogenic tumors, fibrotic tumors and reaches target cancer cells Limiting the amount of lead to low efficacy. Currently, approaches to overcome these delivery barriers for nanotherapeutics and low molecular weight drugs are limited. Previously, the inventors have found that relaxin can improve transport through the tumor matrix but cannot facilitate the delivery of low MW drugs (B. Seed and RK Jain. “Methods to Potentiate Cancer”). Therapies, US Pat. No. 6,719,977, April 13, 2004).

  Presented herein are classes of agents approved by the FDA that can normalize the tumor microenvironment and improve the delivery of both low and high molecular weight drugs. Specifically, the inventors have shown that angiotensin inhibition “normalizes” the stromal matrix in solid tumors, including breast and pancreatic tumors (FIG. 17A). The present inventors have modified the tumor microenvironment through an FDA-approved angiotensin receptor blocker (ARB) and angiotensin converting enzyme inhibitor (ACE-I) to enhance drug delivery through this mechanism. It was assessed whether it was possible. We found that ARB and ACE-I decompressed blood vessels to improve perfusion (Figures 17B-17D) and increased tumor hydraulic conductivity to repair vascular function (Figure 18B). ), And also decided to reduce the stromal matrix density to enhance nanotherapeutic agent penetration (FIG. 18D). These agents improve delivery of molecules as small as oxygen, a radiochemical sensitizer, through vascular normalization (FIGS. 18A-18B) and also enhance penetration of larger agents through stromal matrix normalization ( 18C-18D). Through this repair of the entire tumor microenvironment, these agents enhance the effectiveness of low molecular weight chemotherapeutic and nanotherapeutic drugs in models of breast and pancreatic cancer, resulting in reduced tumor growth and longer animal survival ( Figures 19A-19E). We have combined ARB and ACE-I with all classes of anticancer agents, including small molecule chemotherapeutic agents, biologics, and nanoparticle therapy, which can enhance the delivery of therapeutic agents It has been shown to have wide applicability for treatment.

  Angiotensin blockers show a number of advantages over other approaches. Anti-angiogenic treatment normalizes only the vasculature and is approved only for a limited number of indications. On the other hand, ARB and ACE-I are approved by the FDA as antihypertensives with manageable adverse effects. Matrix degrading enzymes that can normalize the collagen matrix are not selective for tumors and may increase invasion and metastasis. ARB and ACE-I generally do not have the complications associated with matrix remodeling in normal tissues and are therefore safe as antihypertensive drugs. The small molecule drugs ARB and ACE-I are further mediated through nanovectors (eg liposomes, nanoparticles) containing chemotherapeutic drugs to further limit their toxicity and enhance their localization to the tumor May be delivered. Anti-angiogenic drugs, the only FDA-approved aid that enhances drug delivery to tumors, may generally reduce the size of the “pores” in the vessel wall and therefore deliver on larger particles It cannot be improved. Conversely, the angiotensin blockers presented herein can improve delivery for all classes of anti-tumor diagnostic and anti-tumor therapeutic agents.

Example 8: In vitro screening to identify antihypertensive agents for lowering collagen in solid tumors This example is for ranking antihypertensive (AH) agents based on their ability to reduce collagen I levels in tumors An assay is provided.

  Since most collagen I is produced by cancer-associated fibroblasts (CAF), those skilled in the art will recognize that its molecular determinants [active TGFβ1, thrombospondin 1 (TSP1) in CAF supernatant after AH treatment ), And connective tissue growth factor (CTGF)], the level of collagen I can be measured.

  For example, the inventors have determined that losartan reduces TGFβ1 activation and collagen I production in breast CAF in vitro. Cells were treated with 10 μmol / L losartan for 24 hours. Losartan reduced active TGFβ1 levels by 90% (p <0.05), but total TGFβ1 levels were not affected. There was a 27% reduction in the corresponding collagen I level (p <0.05) (see FIG. 1).

Exemplary experimental design:
Antihypertensive: Any FDA-approved angiotensin receptor blocker (ARB) can be tested. Exemplary names and dosages for these agents can be found through, but are not limited to, http://www.globalrph.com/druglist.htm.

  Angiotensin converting enzyme inhibitors (ACEI) also reduce collagen, but they do not target receptors on cells, so we did not measure their effects on collagen I. Calcium channel blockers can also be evaluated for collagen lowering effects.

Cell culture Isolation of cancer-associated fibroblasts (CAF) from human cancer biopsies using a previously described protocol (Orimo A, et al. (2005) Cell 121 (3): 335-348) To do. CAF is seeded in a 24-well plate at a concentration of 500K cells / well and allowed to attach to the plate for 24 hours before adding an antihypertensive drug. For example, all losartan studies were performed at 10 μmol / l for 24 hours based on published protocols (Schuttert JB, et al. (2003) Pflugers Arch 446 (3): 387-393). Treatment can be performed in low serum to reduce background collagen levels. Conditioned media can be collected at the end of the 24 hour treatment period and analyzed for total TGFβ1, active TGFβ1, TSP-1, CTGF, and collagen levels.

Protein Assay In the Losartan study (Schuttert JB, et al. (2003) Pflugers Arch 446 (3): 387-393), collagen I measurements were performed using a type I C-terminal collagen propeptide enzyme-linked immunosorbent assay (ELISA) kit (Quidel , San Diego, CA) and Sircol soluble collagen assay (Biocolor Ltd., United Kingdom). The TGFβ1 assay was performed using a human TGFβ1 ELISA kit (R & D Systems, Minneapolis, Minn.). The assay measures only free mature TGFβ1. To measure the total level of TGFβ1, latent TGFβ1 was activated with 1N HCl. The TSP-1 assay was performed using a human TSP-1 ELISA kit (R & D Systems, Minneapolis, Minn.). The CTGF ELISA kit can be purchased from Leinco (www.leinco.com).

Protocol overview:
Isolate or purchase cancer-associated fibroblasts (CAF) from breast cancer, pancreatic cancer, and colon cancer;
Co-culturing CAF and cancer cells in a medium containing angiotensin I and ACE;
For example, treat CAF with 6 doses of AGTR1 or ACE inhibitors for 48 hours;
Supernatants are collected and TGFβ1, connective tissue growth factor (CTGF), thrombospondin 1 and / or collagen I are measured by ELISA. ELISA measurements should be repeated 3 times or more.

Additional exemplary tests:
(A) Confirm in vitro findings in vivo in a limited number of tumor models.
(B) To screen for new AH, identify the properties of AH to be a more effective modifier of collagen.

Example 9: Combination of Angiotensin Inhibition with Inhibition of Another Fibrotic Pathway to Improve Drug Delivery to Tumors We have shown that normalization of the stromal matrix through angiotensin signaling inhibition is at least partially In particular, it has been found to improve drug delivery through two mechanisms: relaxation of the inherent compressive force in the tumor to improve vascular perfusion, and viscoelastic damage to drug transport directly imparted by the matrix and Reduces steric hindrance. Inhibition of angiotensin signaling results in these changes and can safely inhibit downstream profibrotic TGFβ and CTGF pathway activation. In some embodiments, angiotensin blockers with inhibitors of the pro-fibrotic pathway independent of TGFβ and CTGF activation, including endothelin-1, PDGF, Wnt / βcatenin, IGF-1, TNFα, and IL-4 Can enhance these effects and further improve drug delivery and effectiveness. For example, an endothelin receptor blocker (ERB) and a PDGF inhibitor (PDGF-I) can be used in combination with an angiotensin blocker. ERB treats pulmonary arterial hypertension and can be used as a therapeutic class for cancer, for example, with angiotensin blockers (Nelson et al. (2003) Nature Reviews Vol. 3: 110-116). PDGF-I has been reported for potential antivascular effects in tumors (Baluk et al. (2005) Current Opinion in Genetics & Development 15: 102-111, Andrae et al. (2008) Genes & Development 22 : 1276-1312). Endothelin inhibition occurs through inhibition of TGFβ synthesis (Ogata et al. (2002) Clinical Science 103 (Suppl. 48): 284S-288S) and liver (Binder et al. (2009) Mol. Cancer Ther. 8: 2452-2460). ), Lung (Park et al. (1997) Am J. Respir Crit Care Med Vol. 156: 600-608), and heart have been reported to reduce fibrosis, and tumor progression and metastasis in tumor models (Nelson et al. (Supra); Binder et al. (Supra)). On the other hand, PDGF inhibition has been reported to prevent fibrosis in idiopathic pulmonary fibrosis and scleroderma (Grimminger et al. (2010) Nature Reviews Vol. 9: 956-970; Andrae et al. (2008) Genes & Development 22: 1276-1312), we have determined that it can reduce collagen levels in tumors (data not shown). ERB has been reported to be well tolerated and includes prostate cancer (James et al. (2009) European Urology 55: 1112-1123) and non-small cell lung cancer (Chiappori et al. (2008) Clin Cancer Res 14: 1464-1469) has the potential to improve overall survival. Thus, the combination of angiotensin inhibition with the inhibition of endothelin 1 and / or PDGF by careful dosing should additively improve drug delivery with minimal additional toxicity. In some embodiments, endothelin 1 and / or PDGF at non-medical doses in combination with angiotensin inhibition that can be used at non-hypertensive doses and / or non-anti-tumor doses for improved drug delivery and / or treatment of cancer. Inhibition of can be used.

Equivalents Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Sequence listing:

Claims (70)

Identifying a subject as needing antihypertensive and / or receipt of a collagen modifying agent ("AHCM") based on the need for improved delivery or efficacy of cancer treatment; and (a) AHCM in said subject A method of improving the delivery or efficacy of a cancer treatment in a subject, comprising either or both of: (b) administering a cancer treatment comprising:
The method wherein the AHCM is administered at a dosage sufficient to improve the delivery or efficacy of the cancer treatment. Identifying a subject as needing antihypertensive and / or receipt of a collagen modifying agent ("AHCM") based on the need for improved delivery or efficacy of cancer treatment; and (a) AHCM in said subject A method of treating or preventing cancer in a subject comprising either or both of: (b) administering a cancer therapy comprising:
A method wherein AHCM is administered at a dosage sufficient to treat or prevent cancer. (A) 1H, 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 45nm, 50nm, 75nm, 100nm, 150nm, 200nm as an entity with a hydrodynamic diameter smaller than 300nm as AHCM, cancer treatment Or both are administered;
(B) the subject has not been administered a dose of AHCM within 5 days, within 10 days, within 30 days, within 60 days, or within 100 days of the start of cancer diagnosis or AHCM medication;
(C) the subject is not hypertensive or hypertensive prior to administration of AHCM;
(D) AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier That;
(E) AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier And (f) AHCM is at least 1 hour, 5 hours, 10 hours, or 24 hours; at least 2 days, 5 days, 10 days, or 14 days; at least 2 weeks , 3 weeks, 4 weeks, 5 weeks, or 6 weeks; at least 2 months, 3 months, 4 months, 5 months, or 6 months; or a period of at least 1 year, 2 years, 3 years, 4 years, or 5 years 3. The method of claim 1 or 2, comprising one or more of being administered continuously over time. AHCM
(I) angiotensin II receptor blocker (AT ₁ blocker),
(Ii) renin angiotensin aldosterone antagonists (“RAAS antagonists”),
(Iii) angiotensin converting enzyme (ACE) inhibitor,
(Iv) thrombospondin 1 (TSP-1) inhibitor,
4. The method according to claim 3, which is selected from one or more of (v) a transforming growth factor β1 (TGF-β1) inhibitor or (vi) a connective tissue growth factor (CTGF) inhibitor. AHCM is losartan (COZAAR (registered trademark)), candesartan (ATACAND (registered trademark)), eprosartan mesylate (TEVETEN (registered trademark)), EXP 3174, irbesartan (AVAPRO (registered trademark)), L158,809, olmesartan (BENICAR (registered trademark)), salaracin, telmisartan (MICARDIS (registered trademark)), valsartan (DIOVAN (registered trademark)), or an AT ₁ inhibitor selected from one or more of these derivatives, The method of claim 3.   4. The method of claim 3, wherein AHCM is losartan.   AHCM is selected from one or more of aliskiren (TEKTURNA®, RASILEZ®), remixylene (Ro 42-5892), enalkylene (A-64662), SPP635, or derivatives thereof 4. The method of claim 3, wherein the method is a RAAS antagonist.   AHCM is Benazepril (LOTENSIN (registered trademark)), Captopril (CAPOTEN (registered trademark)), Enalapril (VASOTEC (registered trademark)), Hoshinopril (MONOPRIL (registered trademark)), Lisinopril (PRINIVIL (registered trademark), ZESTRIL (registered trademark) Trademark)), moexipril (UNIVASC (registered trademark)), perindopril (ACEON (registered trademark)), quinapril (ACCUPRIL (registered trademark)), ramipril (ALTACE (registered trademark)), trandolapril (MAVIK (registered trademark)) 4. The method according to claim 3, which is an ACE inhibitor selected from one or more of the derivatives thereof.   4. The method according to claim 3, wherein the AHCM is a TSP-1 inhibitor selected from one or more of ABT-510, CVX-045, LSKL, or derivatives thereof.   4. The method according to claim 3, wherein the TGF-β1 inhibitor is selected from one or more of an anti-TGF-β1 antibody or a TGF-β1 peptide inhibitor.   4. The method of claim 3, wherein the CTGF inhibitor is selected from one or more of DN-9693, FG-3019, or derivatives thereof.   5. The method of claim 4, wherein AHCM is administered in an amount sufficient to enhance the dispersion or efficacy of the cancer treatment.   AHCM may reduce the level or production of collagen, decrease tumor fibrosis, increase stromal tumor transport, improve tumor perfusion, or enhance penetration or spread of cancer treatment in the tumor or tumor vasculature in a subject 5. The method of claim 4, wherein the method is administered at a dose that causes one or more of:   4. The method of claim 3, wherein the AHCM is losartan and is administered at 25-100 mg / day.   4. The method of claim 3, wherein the AHCM is losartan and is provided in a 12.5 mg, 25 mg, 50 mg, or 100 mg dosage form.   AHCM is losartan, 0.25-17.5 mg / day, 0.5-15 mg / day, 1.3-12 mg / day, 1.5-12 mg / day, 2-12 mg / day, 2-10 mg / day, 2-5 mg / day, 4. The method of claim 3, wherein the method is administered at a sub-anti-hypertisive dose in the range of 2-3 mg / day or 2 mg / day.   AHCM is losartan, a dose that is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times higher than the standard medical dose used for antihypertensive or anti-heart failure, or 4. The method of claim 3, wherein the method is administered at the above dose.   5. The method of claim 4, wherein AHCM is administered as a first course of treatment with a non-hypertensive dose of AHCM and then administered as a second course of treatment with AHCM above the standard hypertensive dose.   AHCM is administered as an entity having a hydrodynamic diameter greater than 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm. Item 4. The method according to Item 4.   20. The method of claim 19, wherein AHCM is administered as a polymer nanoparticle or lipid nanoparticle.   Cancer administered as an entity with a hydrodynamic diameter greater than 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm but less than 300 nm 5. The method of claim 4, which is a therapeutic agent.   24. The method of claim 21, wherein the cancer therapeutic is administered as polymer nanoparticles or lipid nanoparticles.   AHCM or cancer therapeutic agent each independently has a hydrodynamic diameter of 1 nm or less or 0.1 to 1.0 nm; a hydrodynamic diameter of 5 to 20 nm or 5 to 15 nm; or 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 5. The method of claim 4, provided as an entity having a size range (nm) of hydrodynamic diameter of 30 nm, 35 nm, 45 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, but less than 300 nm. Subject
(I) not having hypertension,
5. One or more of (ii) not being treated for hypertension at the start of AHCM treatment; or (iii) having normal or low blood pressure Method.   5. The method of claim 4, wherein the subject has not been administered AHCM within 5 days, within 10 days, within 30 days, within 60 days, or within 100 days of diagnosing cancer or initiating AHCM dosing.   5. The method of claim 4, wherein the subject is in need of cancer treatment or is being considered for cancer treatment.   5. The method of claim 4, comprising determining whether the subject has cancer and administering AHCM and a cancer treatment in response to the determination.   5. The method of claim 4, wherein the subject is at risk of having a solid fibrous tumor or has a solid fibrous tumor.   30. The method of claim 28, wherein the subject has a preneoplastic condition or is predisposed to cancer.   Cancer is pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, prostate cancer, cervical cancer, gastrointestinal cancer, gastric cancer, head and neck cancer, kidney cancer, or liver cancer, or metastatic lesions thereof 5. The method according to claim 4, wherein the method is selected from one or more of them.   5. The method of claim 4, wherein the AHCM is administered prior to and / or in combination with a cancer treatment.   32. The method of claim 31, wherein the cancer treatment is selected from one or more of anti-cancer agents, surgery, and / or radiation.   AHCM is administered at least 1 day, 2 days, 3 days, or 5 days before cancer treatment; or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or earlier Item 33. The method according to Item 32.   35. The method of claim 32, wherein the AHCM is maintained for a preselected portion of the time that the subject receives cancer treatment.   35. The method of claim 34, wherein the AHCM is maintained for the entire period during which the cancer treatment is administered.   AHCM is at least 1 hour, 5 hours, 10 hours, or 24 hours; at least 2 days, 5 days, 10 days, or 14 days; at least 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks; at least 2 weeks 5. The method of claim 4, wherein the method is administered continuously over a period of months, 3 months, 4 months, 5 months, or 6 months; or at least 1 year, 2 years, 3 years, 4 years, or 5 years.   5. The method of claim 4, wherein AHCM is administered as a sustained release formulation.   5. The method of claim 4, wherein the AHCM is formulated for oral delivery, subcutaneous delivery, or intravenous continuous delivery.   5. The method of claim 4, wherein AHCM is administered via a subcutaneous pump, implant, or depot. Cancer treatment
(I) a viral cancer therapeutic agent, an anticancer therapeutic agent lipid nanoparticle, an anticancer therapeutic agent polymer nanoparticle, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a cancer therapeutic agent selected from a chemotherapeutic agent,
(Ii) radiation,
5. The method according to claim 4, wherein the method is selected from one or more of (iii) surgery, or (iv) any combination of (i) to (iii).   41. The method of claim 40, wherein the lipid nanoparticles are selected from PEGylated liposomal doxorubicin (DOXIL®) or liposomal paclitaxel (eg, Abraxane®).   41. The method of claim 40, wherein the chemotherapeutic agent is selected from gemcitabine, cisplatin, epirubicin, 5-fluorouracil, paclitaxel, oxaliplatin, or leucovorin.   41. The method of claim 40, wherein the antibody against the cancer target is selected from antibodies against HER-2 / neu, HER3, VEGF, or EGFR.   Cancer treatment is a tyrosine kinase inhibitor selected from sunitinib, erlotinib, gefitinib, sorafenib, icotinib, lapatinib, neratinib, vandetanib, BIBW 2992, or XL-647, or cetuximab, panitumumab, saltumumab, nimotuzumab, nimotuzumab, 5. The method according to claim 4, wherein the method is an anti-EGFR antibody.   41. The method of claim 40, wherein the chemotherapeutic agent is a cytotoxic agent or a cell division inhibitor.   41. The method of claim 40, wherein the chemotherapeutic agent is selected from a microtubule inhibitor, a topoisomerase inhibitor, a taxane, an antimetabolite, a mitotic inhibitor, an alkylating agent, or an intercalating agent.   5. The method of claim 4, wherein the cancer treatment is selected from one or more of anti-angiogenic agents, vascular targeting agents, or vascular disrupting agents.   AHCM or cancer treatment is administered to a subject by systemic administration selected from oral, parenteral, subcutaneous, intravenous, rectal, intramuscular, intraperitoneal, intranasal, transdermal, or inhalation, or intracavitary introduction, 5. A method according to claim 4. Tumor size;
The level or signaling of one or more of transforming growth factor β1 (TGFβ1), connective tissue growth factor (CTGF), or thrombospondin-1 (TSP-1);
Tumor collagen I level;
Fiber content;
Interstitial pressure;
A plasma or serum biomarker selected from collagen I, collagen III, collagen IV, TGFβ1, CTGF, or TSP-1;
The level of one or more cancer markers;
Emergence of new lesions, metabolism, rate of hypoxia generation;
The appearance of new disease-related symptoms;
Size of tissue mass;
Amount of disease-related pain;
Further monitoring the subject for changes in one or more of histological analysis, lobular pattern, and / or presence or absence of mitotic cells; or tumor invasiveness, primary tumor angiogenesis, or metastatic spread 5. The method of claim 4, comprising.   A pharmaceutical composition comprising nanoparticles comprising AHCM.   A pharmaceutical composition comprising nanoparticles comprising AHCM and a cancer therapeutic agent.   51. The pharmaceutical according to claim 50, wherein the cancer therapeutic agent is selected from a viral cancer therapeutic agent, an anticancer agent lipid nanoparticle, an anticancer agent polymer nanoparticle, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a chemotherapeutic agent. Composition.   52. The pharmaceutical composition according to any one of claims 49 to 51, wherein the nanoparticles are polymer nanoparticles or lipid nanoparticles.   52. The pharmaceutical composition according to any one of claims 49 to 51, wherein the AHCM is formulated in a dosage form according to the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure.   AHCM is 0.01 times, 0.02 times, 0.03 times, 0.04 times, 0.05 times, 0.06 times, 0.07 times, 0.08 times, 0.09 times, 0.1 times the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure, 52. The pharmaceutical composition according to any one of claims 49 to 51, formulated in a dosage form smaller than 0.15 times, 0.16 times, 0.2 times, 0.3 times, 0.4 times, 0.5 times, 0.6 times, 0.7 times.   AHCM is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times greater than or greater than the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure 52. The pharmaceutical composition according to any one of claims 49 to 51, which is formulated in a dosage form.   AHCM is 0.01 times, 0.02 times, 0.03 times, 0.04 times, 0.05 times, 0.06 times, 0.07 times, 0.08 times, 0.09 times, 0.1 times the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure, AHCM dosage form formulated in dosage forms smaller than 0.15 times, 0.16 times, 0.2 times, 0.3 times, 0.4 times, 0.5 times, 0.6 times, 0.7 times.   AHCM is 1.1 times, 1.5 times, 1.7 times, 2 times, 3 times, 4 times, 5 times, 10 times greater than or greater than the standard medical dosage form of AHCM used for antihypertensive or anti-heart failure AHCM dosage form formulated in dosage form. A method of optimizing access to an agent, e.g. a diagnostic or imaging agent, or delivery to a cancer, comprising
Administering a hypotensive and / or collagen modifying agent (“AHCM”) to the subject; and optionally administering the agent to the subject;
(A) the diagnostic or imaging agent has a hydraulic diameter greater than 1 nm, 5 nm, or 20-150 nm;
(B) the agent is a radiation agent, NMRA agent, contrast agent; or (c) the subject is at least 2 days, 3 days, or 5 days, or 1 week, 2 weeks prior to administration of the agent. Or one or more of being treated with AHCM administration initiated prior to administration of the agent for 3 weeks, 4 weeks, 5 weeks, or longer. Contacting cancer or cancer-related cells with a candidate agent;
A method or assay for identifying antihypertensive and / or collagen modification (AHCM) comprising detecting a change in a cancer cell in the presence or absence of the candidate agent,
A method or assay wherein the detected change comprises one or more of increased or decreased active TGFβ, TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen (eg, collagen 1) levels. Candidate drugs include renin angiotensin aldosterone antagonist (“RAAS antagonist”), angiotensin converting enzyme (ACE) inhibitor, angiotensin II receptor blocker (AT ₁ blocker), thrombospondin 1 (TSP-1) inhibitor 60. The method or assay of claim 59, wherein the method or assay is selected from one or more of a transforming growth factor β1 (TGF-β1) inhibitor, or a connective tissue growth factor (CTGF) inhibitor.   61. The method or assay of claim 59 or 60, wherein the candidate agent decreases one or more of activated TGFβ, TGFβ1 levels, connective tissue growth factor (CTGF) levels, or collagen levels.   61. The method or assay of claim 59 or 60, further comprising the steps of comparing the treated method or assay with a reference value and comparing the difference between the treated value and the reference value.   61. The method or assay of claim 59 or 60, wherein said method or assay is performed in vitro, in vivo, or a combination of both.   Evaluating the candidate drug in vitro by adding the candidate drug to the culture medium and analyzing the conditioned medium for an increase or decrease in active TGFβ, TGFβ1 level, connective tissue growth factor (CTGF) level, or collagen level. 61. The method or assay of claim 59 or 60 comprising.   61. The method comprising administering a candidate agent to an animal tumor model; and analyzing the subject for an increase or decrease in active TGFβ, TGFβ1 level, connective tissue growth factor (CTGF) level, or collagen level. Methods or assays.   Antihypertensive and / or collagen modified (AHCM) compositions for use in the treatment of cancer, alone or in combination with cancer therapeutics, or hypotensive and / or collagen modified (AHCM), alone or in combination with cancer therapeutics Of use for treating cancer.   A therapeutic kit comprising antihypertensive and / or collagen modification (AHCM), alone or in combination with a cancer therapeutic, and instructions for the treatment of cancer.   A diagnostic kit comprising an antihypertensive and / or collagen modification (AHCM) alone or in combination with an imaging agent and instructions for the diagnosis of cancer. Selecting a subject as in need of receiving AHCM based on the need for improved delivery or efficacy of the cancer treatment; and (a) administering AHCM to said subject; or (b) cancer treatment. A method of selecting a subject that receives an antihypertensive and / or collagen modifying agent (AHCM) comprising either or both of the steps of administering comprising:
The method wherein the AHCM is administered at a dosage sufficient to improve the delivery or efficacy of the cancer treatment.

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