EP2866791A1 - Novel compositions and uses of anti-hypertension agents for cancer therapy - Google Patents
Novel compositions and uses of anti-hypertension agents for cancer therapyInfo
- Publication number
- EP2866791A1 EP2866791A1 EP20130788513 EP13788513A EP2866791A1 EP 2866791 A1 EP2866791 A1 EP 2866791A1 EP 20130788513 EP20130788513 EP 20130788513 EP 13788513 A EP13788513 A EP 13788513A EP 2866791 A1 EP2866791 A1 EP 2866791A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- ahcm
- cancer
- agent
- therapy
- tumor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- pegylated liposomal doxorubicin DOXIL ®
- DOXIL ® pegylated liposomal doxorubicin
- oncolytic viruses currently in multiple clinical trials, represent two nanotherapeutics whose size ( ⁇ 100 nm) hinders their intratumoral distribution and therapeutic effectiveness (Nemunaitis J, et al. (2001) J Clin Oncol 19:289-298).
- Fibrotic tumors typically have a dense collagen network, which causes small interfibrillar spacing in the interstitium to retard 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) These barriers limit the amount of drug that reaches the target cancer cells and leads to poor drug effectiveness.
- nanotherapeutics e.g., lipid- or polymeric nanoparticles and viruses
- protein and nucleic acid drugs small molecule chemotherapeutic agents and immune cells.
- the invention is based, in part, on the discovery that losartan, an angiotensin II receptor antagonist drug approved for the treatment of high blood pressure (hypertension), improves the delivery and efficacy of cancer therapeutics.
- losartan normalizes the collagen, interstitial matrix of solid tumors and facilitates the distribution and/or penetration of
- chemotherapeutics including large molecular weight chemotherapeutics, e.g.,
- losartan reduced collagen I levels in (e.g., reduced collagen production by) carcinoma associated fibroblasts (CAFs) isolated from breast cancer biopsies, and caused a dose-dependent reduction in stromal collagen in desmoplastic models of human breast, pancreatic and skin tumors in mice.
- Losartan also improved the distribution, therapeutic efficacy and/or penetration of nanopartices (e.g., oncolytic herpes simplex viruses (HSV) and pegylated liposomal doxorubicin (DOXIL ® )).
- HSV oncolytic herpes simplex viruses
- DOXIL ® pegylated liposomal doxorubicin
- losartan facilitates decompression of blood vessels and vascular normalization, and improves tumor perfusion and delivery of low molecular weight chemotherapeutics and oxygen, thus enhancing the therapeutic effect of cancer therapies, including but not limited to radiation, photodynamic therapy, chemotherapeutics and immunotherapies.
- Examples disclosed herein further demonstrate a reduction in collagen levels and tumor solid stress using angiotensin inhibitors other than losartan, including, for example, angiotensin receptor blockers (ARBs), such as candesartan and valsartan, as well as angiotensin converting enzyme inhibitors (ACE-I), such as lisinopril.
- ARBs angiotensin receptor blockers
- ACE-I angiotensin converting enzyme inhibitors
- compositions for improving the delivery and/or efficacy of therapeutics are disclosed.
- Methods and compositions for treating or preventing a cancer e.g., a solid tumor such as a desmoplastic tumor
- an anti-hypertensive and/or collagen modifying agent as a single agent or in combination with a microenvironment modulator, and/or a therapeutic agent
- a cancer therapeutic agent for example, a cancer therapeutic agent ranging in size from an immune cell or a large nanotherapeutic to a low molecular weight chemotherapeutics and/or oxygen radicals
- the invention features a method of treating or preventing a disorder, e.g., a hyperproliferative disorder (e.g., a cancer) in a subject, or of improving the delivery and/or efficacy of a therapy (e.g., a cancer therapy) to a subject.
- a disorder e.g., a hyperproliferative disorder (e.g., a cancer) in a subject
- a therapy e.g., a cancer therapy
- AHCM anti-hypertensive and/or a collagen modifying agent
- administering e.g., the cancer therapy
- the therapy e.g., the cancer therapy
- AHCM and anti-cancer agent under conditions, e.g., of dosage of AHCM and anti-cancer agent, sufficient to treat or prevent the disorder (e.g., the cancer or tumor), in the subject, or to improve the delivery and/or efficacy of the therapy (e.g., the cancer therapy) provided to the subject.
- the disorder e.g., the cancer or tumor
- the therapy e.g., the cancer therapy
- the method includes one or more of the following:
- AHCM AHCM
- microenvironment modulator e.g., the cancer therapy
- therapy e.g., the cancer therapy
- any combination thereof as an entity having a hydrodynamic diameter of greater than about 1, 5, 10, 15, 20, 25, 30, 35, 45, 50, 75, 100, 150, 200 nm, but less than 300 nm, e.g., as a nanoparticle;
- the subject has a history of treatment (or lack of treatment) for hypertension, as described herein, e.g., the subject has not been administered a dose of an AHCM, e.g., an AHCM named herein, or any AHCM (e.g., either of a dose sufficient to substantially lower the subject's blood pressure or a sub-anti-hypertensive dose) , within 5, 10, 30, 60 or 100 days of the diagnosis of cancer or the initiation of the AHCM dosing.
- the subject is not hypertensive, or has been hypertensive, prior to administration of the AHCM;
- a dosing regimen described herein e.g., administration of the AHCM and/or the microenvironment modulator is initiated prior to the initiation of administration of the cancer therapy, e.g., it is initiated at least one, two, three, or five days, or one, two, three, four, five or more weeks prior to cancer therapy (e.g., the AHCM and/or the microenvironment modulator is administered at a minimum of two weeks prior to cancer therapy);
- a dosing regimen described herein e.g., providing a first course of treatment with an AHCM at a sub-anti-hypertensive dose followed by a second, higher dose, course of treatment with an AHCM, e.g., at a dose that is at or above a standard antihypertensive dose (e.g., wherein the second course is administered in a time course that will counteract a
- AHCM and/or the microenvironment modulator substantially continuously over a period of at least 1, 5, 10, or 24 hours; at least 2, 5, 10, or 14 days; at least 2, 3, 4, 5 or 6 weeks; at least 2, 3, 4, 5 or 6 months; or at least 1, 2, 3, 4 or 5 years, or longer;
- AHCM AHCM
- the microenvironment modulator sequentially and/or concurrently with the therapy, e.g., the cancer therapy.
- the microenvironment modulator and the therapy can be administered (at the same or different dosages) in any order and/or overlap with the therapy.
- the AHCM and/or the microenvironment modulator is administered before the therapy (e.g., as described in step d)).
- the AHCM and/or the microenvironment modulator is administered sequentially and/or concurrently with the therapy (e.g., the AHCM and/or the microenvironment modulator is administered prior to the therapy (e.g., as described in step d) and concurrently with the therapy).
- the therapy is administered first, and the AHCM and/or the microenvironment modulator is administered after initiation of the therapy, or is administered after cessation of the therapy.
- the administration of the AHCM and/or the microenvironment modulator starts after cessation of the therapy.
- microenvironment modulator continues after cessation of the therapy.
- administration of the AHCM and/or the microenvironment modulatorand the therapy is concurrent
- the administration of the AHCM, the microenvironment modulator and the therapy can be continued as clinically appropriate, for example, (i) as a combination therapy, (ii) with a period of therapy with either the AHCM or the therapy, or (iii) as a combination of (i) and (ii) in any order.
- the AHCM and/or the microenvironment modulator alters
- the AHCM does not inhibit or prevent (e.g., is administered in an amount insufficient to inhibit or prevent) tumor growth by itself, but sufficient to alter (e.g., enhance) the distribution or efficacy of the therapy, e.g., the cancer therapy.
- the AHCM results in (e.g., is administered at a dose that causes), one or more of: decreases the level or production of an extracellular matrix component, such as a fiber (e.g., collagen, procollagen), and/or a polysaccharide (e.g., a glycosaminoglycan such as hyaluronan or hyaluronic acid); decreases the level or production of collagen or procollagen; decreases the level or production of hyaluronic acid; decreases tumor fibrosis; increases interstitial tumor transport; improves tumor perfusion; increases tumor oxygenation; decreases tumor hypoxia; decreases tumor acidosis; enables immune cell infiltration; decreases immunosuppression; increases antitumor immunity; decreases the production of cancer stem cells (also referred to herein as tumor- initiating cells); or enhances the efficacy (e.g., penetration or diffusion), of the therapy, e.g., the cancer therapy (e.g., the cancer therapy (e.g
- the AHCM and/or microenvironment modulator is administered in a dosage sufficient to improve the delivery or effectiveness of the therapy.
- the method results in, or comprises (e.g., the AHCM and/or microenvironment modulator is administered in a dosage sufficient to result in) improvement of a disorder-related parameter in said subject, as compared to a subject treated with said therapy but without administration of the AHCM and/or microenvironment modulator.
- Disorder-related parameter refers to a parameter that varies with the alleviation of the disorder or a symptom of the disorder.
- an AHCM (and, in embodiments not a microenvironment modulator) is administered and the improvement is as compared to a subject treated with said therapy but without administration of the AHCM.
- a microenvironment modulator (and, in embodiments not an AHCM) is administered and the improvement is as compared to a subject treated with said therapy but without administration of the microenvironment modulator.
- an AHCM and a microenvironment modulator are administered and the improvement is as compared to a subject treated with said therapy but without administration of the AHCM and the microenvironment modulator.
- the parameter comprises relief of a symptom of said disorder.
- the parameter comprises outcome of a patient scored evaluation of symptoms or quality of life, e.g., a quality of life questionaire, e.g., outcome on an evaluation of number of meals consumed on the day prior to the evaluation, pain, weight loss or gain.
- a quality of life questionaire e.g., outcome on an evaluation of number of meals consumed on the day prior to the evaluation, pain, weight loss or gain.
- the parameter comprises one or more or all of:
- the parameter comprises one or more or all of: a) drug concentration, e.g., at a disorder or disease site, e.g., in a solid tumor; b) tumor response; c) blood perfusion, e.g., at a disorder or disease site, e.g., in a solid tumor; d) oxygenation, e.g., at a disorder or disease site, e.g., in a solid tumor; e) interstitial fluid pressure at a disorder or disease site, e.g., in a solid tumor; or f) extracellular matrix content or composition, e.g., level of collagen , hyaluronic acid.
- drug concentration e.g., at a disorder or disease site, e.g., in a solid tumor
- tumor response e.g., at a disorder or disease site, e.g., in a solid tumor
- c) blood perfusion e.g., at a disorder or disease site, e.
- the parameter is evaluated by a non-invasive method, e.g., a magnetic resonance method, or MRS, PET, or SPECT.
- a non-invasive method e.g., a magnetic resonance method, or MRS, PET, or SPECT.
- the disorder is, e.g., cancer
- said parameter is drug concentration, e.g., at a disorder or disease site, e.g., in a solid tumor.
- the parameter cam be evaluated by a method described herein, e.g., with any of PET-CT, e.g., generally as described in Saleem et al. (2000) The Lancet 355: 2125-2131, MRS, e.g., generally as described in Meisamy et al. (2004) Radiology 233: 424-431, or SPECT, e.g., generally as described in Perik et al. (2006) Journal of Clinical Oncology 24: 2276-2282.
- the disorder is, e.g., cancer
- said parameter is blood perfusion, e.g., at a disorder or disease site, e.g., in a solid tumor.
- the parameter can be evaluated by a method described herein, e.g. MRI, e.g., generally as described in Sorensen et al. (2012) Cancer Research 72: 402-407, or perfusion CT e.g., generally as described in Park et al. (2009) Radiology 250: 1 10-1 17, or Doppler ultrasound generally as described in Singh et al. (2010) European J. of Radiology 75: el58- 162.
- the disorder is, e.g., cancer
- said parameter is oxygenation, e.g., at a disorder or disease site, e.g., in a solid tumor.
- the parameter can be evaluated by a method described herein, e.g., PET, PET-CT, e.g. generally as described in Rajendran et al. (2006) Clinical Cancer Research 12: 5435-5441, or Eppendorf electrode, e.g. generally described in Le et al. (2007) International J. of Radiation Oncology Biology Physics 69: 167-175, or immunohistochemistry, e.g. generally described in Rademakers et al. (201 1) BMC Cancer 11 : 167.
- the disorder is, e.g., cancer
- said parameter is metabolic activity, e.g., at a disorder or disease site, e.g., in a solid tumor
- the parameter can be evaluated by a method described herein, e.g., functional MRI, or PET, PET- MRI, PET-CT, e.g. generally as described in Shankar et al. (2006) The Journal of Nuclear Medicine 47: 1059- 1066.
- the disorder is, e.g., cancer
- said parameter is interstitial fluid pressure, e.g., at a disorder or disease site, e.g., in a solid tumor.
- the parameter can be evaluated by a method described herein, e.g., the wick- in- needle technique, e.g., generally as described in Boucher et al. (1991) Cancer Research 51 : 6691-6694.
- the disorder is a hyperproliferative fibrotic disease and said parameter is amount of connective tissue matrix or blood perfusion.
- the disorder is an inflammatory disorder
- said parameter is amount of connective tissue matrix.
- the parameter can be evaluated immunohistochemically.
- the disorder is an autoimmune disorder
- said parameter is amount of connective tissue matrix.
- the parameter can be evaluated immunohistochemically.
- the parameter is evaluated in a sample from said subject, e.g., a tumor sample, e.g., a biopsy, or a blood or serum sample.
- a tumor sample e.g., a biopsy
- a blood or serum sample e.g., a blood or serum sample.
- the parameter comprises one or more or all of: a) drug concentration, e.g., as evaluated by HPLC, or or NMR, e.g., evaluated generally as described in Olive et al. (2009) Science 324: 1475, HPLC with tandem MS, generally as described in Hu et al. (2011) JNCI 103 : 893-905, or by histological measures, e.g., fluorescence imaging of fluorescent drugs, generally as described in Primeau et al. (2005) Clinical Cancer Research 11 : 8782-8788;
- collagen content e.g., as evaluated by total collagen content measured by hydroxyproline content, e.g., generally as described in Netti et al. (2000) Cancer Research 60: 2497-2503, or immunohistochemistry by antibody staining, e.g., generally as described in Pluen et al. (2001) PNAS 98:4628-4633;
- hyaluronan content e.g., as evaluated by hyaluronan-binding protein labeling of tissue sections, as generally described in Pluen et al. (2001) PNAS 98:4628-4633, or glycosaminoglycan analysis in tissue extracts, e.g., generally as described in Netti et al. (2000) Cancer Research 60: 2497-2503;
- pathological response e.g., the prevalence of tumor cells in a sample, e.g., evaluated generally as described in Minckwitz et al. (2012) Journal of Clinical Oncology published as 10.1200/JCO.2011.38.8595;
- vessel morphology e.g., size
- vessel morphology e.g., size
- hypoxia e.g., generally as described in Rademakers et al. (2011) BMC Cancer 1 1 : 167 or Le et al. (2007) International J. of Radiation Oncology Biology Physics 69: 167- 175.
- Hypoxia can be evaluated in a number of ways, e.g.: by a pimonizadole method, see, e.g., Kaanders, J. H. et al. (2002) Cancer Res. 62, 7066-7074; an EF5 method, see, e.g., Evans, S. M. et al. (2007) Int. J. Radiat. Oncol.Biol. Phys.
- a CA9 method see, e.g., Koukourakis, M. I. et al, . (2006) J. Clin. Oncol. 24, 727-735
- a LOX method see, e.g., Erler, J. T. et al, . (2006) Nature 440, 1222-1226
- a HIF method see, e.g., Bos, R. et al. (2003) Cancer 97, 1573-1581, Yan,et al. (2009) Br. J. Cancer 101, 1 168- 1 174, or Koukourakis, M. I. et al, (2006) J. Clin. Oncol.
- the parameter comprises one or more or all of:
- ICTP serum degraded collagen
- PIP collagen synthesis
- fibrotic factors connective tissue growth factor (CTGF), transforming growth factor-beta (TGF-beta), interleukin-1, -4, -6, -8, - 10 and -13, platelet - derived growth factor (PDGF), stromal cell-derived factor 1 (SDF1), e.g., evaluated generally as described in Harti et al. (2006) American J. of Respiratory Medicine 173 : 1371- 1376.
- CGF connective tissue growth factor
- TGF-beta transforming growth factor-beta
- PDGF platelet - derived growth factor
- SDF1 stromal cell-derived factor 1
- the parameter is drug concentration and said parameter is evaluated by a chromatographic method, e.g., HPLC.
- the disorder is a hyperproliferative fibrotic disease and the parameter is fibrosis.
- the disorder is an inflammatory disorder and the parameter is fibrosis.
- the disorder is an autoimmune disorder and the parameter is fibrosis.
- the parameter is a morphological parameter, e.g., evaluated at a disorder or disease site, e.g., in a solid tumor and comprises one or more or all of:
- vessel diameter or size evaluated e.g., evaluated generally as described in Provenzano et al. (2012) Cancer Cell 21 :418-429.
- the AHCM is chosen from one or more of:
- an angiotensin II receptor blocker (ATi blocker)
- RAAS antagonist an antagonist of renin angiotensin aldosterone system
- ACE angiotensin converting enzyme
- TSP-1 thrombospondin 1
- TGF- ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- SDF- la stromal cell-derived growth factor 1 alpha
- AHCM AHCM
- the method can include one, two, three or more AHCMs, alone or in combination with one or more cancer therapies.
- the AHCM is a RAAS antagonist. In an embodiment, the AHCM is a RAAS antagonist.
- RAAS antagonist is chosen from one or more of: aliskiren (TEKTURNA®, RASILEZ®), remikiren (Ro 42-5892), enalkiren (A-64662), SPP635, or a derivative thereof.
- the AHCM is an ATi inhibitor.
- the ATi blocker is chosen from one or more of: losartan (COZAAR®), candesartan
- the AHCM is an ACE inhibitor.
- the ACE inhibitor is chosen from one or more of: benazepril (LOTENSIN®), captopril (CAPOTEN®), enalapril (VASOTEC®), fosinopril (MONOPRIL®), lisinopril (PRINIVIL®, ZESTRIL®), moexipril (UNIVASC®), perindopril (ACEON®), quinapril (ACCUPRIL®), ramipril (ALTACE®), trandolapril (MAVIK®), or a derivative thereof.
- the AHCM is a TSP- 1 inhibitor.
- the TSP- 1 inhibitor is chosen from one or more of: ABT-510, CVX-045, LSKL, or a derivative thereof.
- the AHCM is a TGF- ⁇ inhibitor, e.g., an anti- TGF- ⁇ antibody, a TGF- ⁇ peptide inhibitor.
- the TGF- ⁇ inhibitor is chosen from one or more of: CAT-192, fresolimumab (GC1008), LY 2157299, Peptide 144 (P144), SB-431542, SD-208, compounds described in U.S. Patent Serial No. 7,846,908 and U.S. Patent Application Publication No. 2011/0008364, or a derivative thereof.
- the AHCM is a CTGF inhibitor.
- the CTGF inhibitor is chosen from one or more of: DN-9693, FG-3019, and compounds described in European Patent Application Publication No. 1839655, U.S. Patent Serial No. 7,622,454, or a derivative thereof.
- the AHCM is an inhibitor of stromal cell-derived growth factor 1 alpha (SDF-la/CXCL12a).
- the SDF- 1 a inhibitor is an anti- SDF la antibody or fragment thereof.
- the SDF- 1 a inhibitor is an inhibitor of an SDF- 1 a receptor (e.g., a CXCR4 inhibitor), for example Plerixafor (AMD- 3100).
- exemplary AHCMs are described herein are not limiting, e.g, derivatives of AHCMs described herein can be used in the methods described herein.
- Methods of the invention use an AHCM to potentiate a therapy ⁇ e.g., a cancer therapy).
- the AHCM is administered at a dose that corresponds to a standard of care dose.
- Standard of care doses of the AHCM are available in the art.
- the AHCM is the ATi inhibitor, losartan
- the standard of care dose for antihypertensive use in a human is about 25- 100 mg day "1 .
- losartan can be administered orally in a daily schedule (once or twice a day), alone or in combination with a cancer therapy described herein.
- Losartan can be provided in a dosage form ⁇ e.g., an oral tablet) of about 12.5 mg, 25 mg, 50 mg or 100 mg.
- Exemplary standard of care doses for other ATi inhibitors for antihypertensive or anti-heart failure use in humans are as follows: 4 to 32 mg day "1 of candesartan (ATACAND®) ⁇ e.g., available in a dosage form for oral administration containing 4 mg, 8 mg, 16 mg, or 32 mg of candesartan); 400 to 800 mg day "1 of eprosartan mesylate (TEVETEN®) ⁇ e.g., available in a dosage form for oral administration containing 400 or 600 mg of eprosartan); 150 to 300 mg day "1 of irbesartan (AVAPRO®) ⁇ e.g., available in a dosage form for oral administration containing 150 or 300 mg of irbesartan); 20 to 40 mg day "1 of olmesartan (BENICAR®) (available in a dosage form for oral administration containing 5 mg, 20 mg, or 40 mg of olmesartan); 20 to 80 mg day "1 of
- MICARDIS® ⁇ e.g., available in a dosage form for oral administration containing of 20 mg, 40 mg or 80 mg of telmisartin
- 80 to 320 mg day "1 of valsartan ⁇ e.g. , available in a dosage form for oral administration containing 40 mg, 80 mg, 160 mg or 320 mg of valsartan).
- the AHCM is administered at a sub-anti-hypertensive dose
- the AHCM is administered in an amount that does not substantially lower the mean arterial blood pressure of the subject, e.g., as measured after a pre-selected number of administrations at that dosage, e.g., at the steady state plasma level for a given dosage.
- the AHCM is administered, at least once, at a dose that reduces mean arterial blood pressure in the subject by less than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%.
- the AHCM is administered at a dose that reduces blood pressure by less than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or less of the reduction caused by a standard of care anti-hypertensive dose for that AHCM.
- the AHCM is administered at a dose that is less than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90% of the dose of that AHCM that would bring the subject's blood pressure into the normal range, e.g, about 120 systolic and about 80 diastolic, or a dose that would bring the subjects blood pressure into the range of to 120+/-5 systolic and 80+/-5 diastolic.
- the AHCM is administered at a dose that is less than the standard of care dose for anti-hypertensive or anti-heart failure use (e.g., a dose that 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, that of the standard of care dose for anti-hypertensive or anti-heart failure use).
- Standard of care doses of the AHCM are available in the art.
- the suboptimal antihypertensive drug can range from 0.25 to 17.5, 0.5 to 15, 1.3 to 12, 1.5 to 12, 2 to 12, 2 to 10, 2 to 5, 2 to 3 mg day "1 , typically, 2 mg day "1 .
- the AHCM is losartan and is administered at a dose less than 25, 20, 15, 10, 5, 4, 3, 2, 1 mg day "1 .
- Losartan can be administered orally in a daily schedule (once or twice a day) at a sub-anti-hypertensive dose of 2-3 mg day "1 , alone or in combination with a cancer therapeutic described herein.
- Exemplary standard of care doses for other ATi inhibitors are as follows: 4 to 32 mg day “1 of candesartan (ATACAND®), 400 to 800 mg day “1 of eprosartan mesylate (TEVETEN®), 150 to 300 mg day “1 of irbesartan (AVAPRO®), 20 to 40 mg day “1 of olmesartan (BENICAR®), 20 to 80 mg day “1 of telmisartin (MICARDIS®), and 80 to 320 mg day "1 of valsartan
- the AHCM is administered at a dose that is less than the standard of care dose of the anti-hypertensive or anti-heart failure dose (e.g., a dose that 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, that of the standard of care dose of the anti-hypertensive or anti-heart failure dose for other ATi inhibitors such as candesartan, eprosartan, irbesartan, olmesartan, telmisartin, and valsartan).
- the standard of care dose of the anti-hypertensive or anti-heart failure dose e.g., a dose that 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, that of
- the AHCM is formulated in a dosage form that is less than the standard of care anti-hypertensive or anti-heart failure dosage form (e.g., a dosage form that is less than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.16, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, that of the standard of care dosage form).
- the dosage form can be of about 0.5 mg- 1 1 mg; 1 mg -10 mg; 1-5 mg, or 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg or 10 mg.
- losartan can be provided in a dosage form (e.g., an oral tablet) below 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 1 1 mg, or about 12 mg.
- a dosage form e.g., an oral tablet below 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 1 1 mg, or about 12 mg.
- the AHCM is formulated and/or dosed for oral administration.
- the AHCM is formulated as a tablet (e.g., an oral tablet). In other words, a tablet (e.g., an oral tablet).
- the AHCM is formulated and/or dosed for other routes of administration, e.g., subcutaneous, intravenous, or intraperitoneal administration.
- the AHCM can be formulated and/or dosed for extended, delayed, or controlled release, e.g., in an extended release formulation (e.g., an oral formulation) for substantially continuous release over a period of hours (e.g., at least 1, 2, 3, 4, 5, 10, or 24 hours); days (e.g., at least 1, 2, 5, 10, 14 days, or longer), weeks, months or years.
- the sub-anti-hypertensive dose of the AHCM or a dose of the AHCM that is less than the standard of care dose for anti-hypertensive or anti -heart failure use can be a dose that is insufficient to inhibit or prevent tumor growth or progression if it is administed to a subject by itself.
- the AHCM is administered at a dose that is greater than the standard of care dose for anti-hypertensive or anti-heart failure use (e.g., a dose that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dose for anti-hypertensive or anti-heart failure use).
- Standard of care doses of the AHCM are available in the art; some of which are exemplified herein.
- the AHCM is formulated in a dosage form that is greater than the standard of care anti -hypertensive or anti-heart failure dosage form (e.g., a dosage form that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dosage form).
- Standard of care dosage forms of the AHCM are available in the art; some of which are exemplified herein.
- a dose of the AHCM that is comparable to, or greater than the standard of care anti -hypertensive or anti-heart failure dose can be a dose that is insufficient to inhibit or prevent tumor growth or progression if it is administed to a subject by itself.
- the anti-cancer agent is administered at a greater dosage, or in a regimen that results in higher levels of the anti-cancer agent, as compared with a reference, e.g., the dosage on a package insert, the standard of care dosing, or the maximum tolerated dose (MTD).
- a reference e.g., the dosage on a package insert, the standard of care dosing, or the maximum tolerated dose (MTD).
- the anti-cancer agent is administered at a lesser dosage, or in a regimen that results in lower levels of the anti-cancer agent, as compared with a reference, e.g., the dosage on a package insert, the standard of care dosing, or the MTD.Jn some embodiments, the anti-cancer agent is administered in an amount such that it is not effective to inhibit or prevent tumor growth or progression when administered by itself, but in an amount sufficient to inhibit or prevent tumor growth or progression when administered in combination with the AHCM.
- the cancer therapy or cancer therapeutic when administered in combination with an AHCM, is administered to the subject at a dose that is less than the lowest dose that would be used in the absence of the AHCM, to treat or prevent cancer in a subject.
- the dose of the anti-hypertenisve and/or collagen modifying agent can be a dose that is less than the lowest dose that would be used to treat a hypertensive-associated disorder or heart failure, while the dose of the cancer therapy or cancer therapeutic can be a dose that is less than the lowest dose that would be used in the absence of the AHCM, to treat or prevent cancer in a subject.
- the dose of the 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
- the dose of the AHCM agent administered to the subject as an adjuvant can be less than the lowest dose that would be used alone to treat cancer.
- the dose of the AHCM agent administered to the subject as an adjuvant can be sub- anti-hypertensive dose or comparable to, or greater than the standard care dose for treatment of hypertension or heart failure.
- the dose of the cancer therapy or cancer therapeutic administered to the subject is less than the lowest dose that would be used in the absence of the AHCM, to treat a patient with cancer
- the dose of the AHCM agent administered to the subject as an adjuvant can be less than the lowest dose that would be used alone to treat cancer, but is sufficient to improve efficacy of a cancer therapy or delivery of a cancer therapeutic to a tumor.
- the dose of the AHCM agent administered to the subject as an adjuvant can be sub-anti -hypertensive dose or comparable to, or greater than the standard care dose for treatment of hypertension or heart failure.
- Methods to determine the lowest dose of any agent, e.g., an anti-cancer agent and/or an AHCM, for treatment are well known within one of skill in the art.
- a skilled artisan can determine the lowest dose of an AHCM and/or an anti-cancer agent effective for treatment in an animal model corresponding to a specific type of cancer, e.g., by administering the animal with different doses of the AHCM and/or anti-cancer agent and monitoring the tumor growth as compared to a control.
- a control can be an animal treated with an anti-cancer agent alone (i.e., in the absence of the AHCM).
- the AHCM and the therapy can be administered in combination, e.g., sequentially and/or concurrently, as described herein.
- the AHCM and the therapy can be administered (at the same or different dosages) in any order and/or overlap with the therapy.
- the AHCM is administered before the therapy.
- the AHCM is administered sequentially and/or concurrently with the therapy (e.g., the AHCM is administered prior to the therapy and concurrently with the therapy).
- the cancer therapy is administered first, and the AHCM is administered after initiation of the cancer therapy, or is administered after cessation of the therapy.
- the administration of the AHCM starts after cessation of the therapy (e.g., with or without a gap between the cessation of the therapy and the beginning of the AHCM). In other embodiments, the administration of the AHCM continues after cessation of the therapy. In embodiments where administration of the AHCM and therapy is concurrent, the administration of the AHCM and the cancer therapy can be continued as clinically appropriate (i) as a combination therapy, (ii) with a period of therapy with either the AHCM or the cancer therapy, or (iii) a combination of (i) and (ii) in any order.
- the administration of the AHCM can be substantially continuous.
- administration of the AHCM can be substantially continuously over a period of at least 1, 5, 10, 24 hours; 2, 5, 10, 14 days, or longer.
- substantially continuous administration of an AHCM e.g., via a subcutaneous pump
- administration e.g., single or multiple subcutaneous administrations of the AHCM.
- the AHCM administration continues after the therapy has ceased, e.g., over a period of hours, days, months or years.
- the administration of the AHCM can be intermittent, e.g., can have gaps at pre-determined intervals, during the course of therapy.
- two or more doses of the AHCM are administered, alone or in combination with the therapy (e.g., the cancer therapy).
- the AHCM is administered at a suboptimal anti-hypertensive dose and an anti-hypertensive dose during the course of therapy.
- a suboptimal anti-hypertensive dose of the AHCM can be
- therapy e.g., cancer therapy
- an anti-cancer agent that increases mean arterial blood pressure e.g, treatment with an anti- angiogenic drug (e.g., Avastin, sunitinib or sorafenib)
- an anti- angiogenic drug e.g., Avastin, sunitinib or sorafenib
- an AHCM is administered as an entity having a hydrodynamic diameter of greater thanabout 1, 5, 10, 100, 500, or 1,000 nm.
- the AHCM can be a protein, e.g., an antibody.
- the AHCM can also be administered as a nanoparticle, e.g., a polymeric nanoparticle or a liposome, that includes the AHCM as a small molecule therapeutic or a protein, e.g., an antibody.
- the therapy is a cancer therapeutic (also referred to herein as "an anti-cancer agent") or second therapeutic agent is administered as an entity having a hydrodynamic diameter of greater than about 1, 5, 10, 20, 50, 75, 100, 150, 200, 500, or 1,000 nm.
- the second therapeutic agent e.g., the anti-cancer agent
- the second therapeutic agent (e.g., the anti-cancer agent) can also be administered as a nanoparticle, e.g., a polymeric nanoparticle or a liposome, that includes the agent as a small molecule therapeutic (i.e., a molecule having a hydrodynamic diameter of about 1 nm or less) or a protein, e.g., an antibody.
- a nanoparticle e.g., a polymeric nanoparticle or a liposome
- the agent i.e., a molecule having a hydrodynamic diameter of about 1 nm or less
- a protein e.g., an antibody
- an AHCM is administered as an entity having a hydrodynamic diameter of greater than about 1 nm (e.g., greater than about 1, 5, 10, 20, 50, 75, 100, 150, 200, 500, or 1,000 nm) and a second therapeutic agent (e.g., an anti-cancer agent) is administered as an entity having a hydrodynamic diameter of about lnm or less.
- a second therapeutic agent e.g., an anti-cancer agent
- the AHCM is present in the entity without a second therapeutic agent (e.g., a chemotherapeutic agent).
- the AHCM can be formulated for extended release, e.g., in an extended release formulation for substantially continuous release for hours, days, weeks, months or years.
- an AHCM is administered as an entity having a hydrodynamic diameter of about 1 nm, or less
- a second therapeutic agent e.g.,w anticancer agent
- a hydrodynamic diameter of about 1 nm or greater e.g., greater than about 1, 5, 10, 20, 50, 75, 100, 150, 200, 500, or 1,000 nm.
- an AHCM is administered as an entity having a hydrodynamic diameter of less than, or equal to, about 1 nm and a second therapeutic agent anti-cancer agent) is administered as an entity having a hydrodynamic diameter of less than about 1 nm.
- an AHCM is administered as an entity having a hydrodynamic diameter of greater than about 1 nm (e.g., greater than about 1, 5, 10, 20, 50, 75, 100, 150, 200, 500, or 1,000 nm)
- a second therapeutic agent e.g., an anti-cancer agent
- the AHCM and the second therapeutic agent can be in separate or the same entity.
- the AHCM can be provided as a first nanoparticle and the second therapeutic agent (e.g., the anti-cancer agent) provided as a second nanoparticle (e.g., where the second nanoparticle has a structural property (e.g., size or composition) or a functional property (e.g., release kinetics or a pharmacodynamic property) that differs from the first nanoparticle).
- the second therapeutic agent e.g., the anti-cancer agent
- a second nanoparticle e.g., where the second nanoparticle has a structural property (e.g., size or composition) or a functional property (e.g., release kinetics or a pharmacodynamic property) that differs from the first nanoparticle).
- an AHCM and a second therapeutic agent e.g., an anti-cancer agent
- the AHCM is selected from a therapeutic entity having a hydrodynamic diameter: equal to or less than 1 or 2 nm; between 2 - 20, 10-25, 20-40, 40, 50-150 nm; between 10, 15, 20, 25, 35, 40, 45, 50- 100 nm; between 10, 15, 20, 25, 35, 40, 45, 50 -200 nm; between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300 -500 nm; and between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300, 1000 nm; or 10, 15, 20,25, 35, 45, 50, 75, 100, 150 or 200 nm.
- the AHCM is a small molecule therapeutic; is a protein, e.g., an antibody; or is provided in a nanoparticle.
- the anti-cancer agent or second therapeutic agent is selected from a therapeutic entity having a hydrodynamic diameter: equal to or less than 1 or 2 nm; between 2 - 20, 10-25, 20-40, 40, 50-150 nm; between 10, 15, 20, 25, 35, 40, 45, 50- 100 nm; between 10, 15, 20, 25, 35, 40, 45, 50 -200 nm; between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300 -500 nm; and between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300 - 1000 nm; or 10, 15, 20,25, 35, 45, 50, 75, 100, 150 or 200 nm.
- the anti-cancer agent or second therapeutic agent is a small molecule therapeutic with a hydrodynamic diameter of 1 nm or less; is a protein, e.g., an antibody; or is provided in a nanoparticle.
- the AHCM, or anti-cancer agent or the second therapeutic agent can be provided as an entity having the following size ranges (in nm): a hydrodynamic diameter of less than or equal to 1, or between 0.1 and 1.0 nm, e.g., that of a typical small molecule; a hydrodynamic diameter of between 5 and 20, or 5 and 15 nm, e.g., that of a protein, e.g., an antibody; or a hydrodynamic diameter of 10-5,000, 20-1, 000, 10-500, 10-200, 10- 150, or 10- 100, 10-25, 20-40, 40, 50- 150 nm; between 10, 15, 20, 25, 35, 40, 45, 50- 100 nm; between 10, 15, 20, 25, 35, 40, 45, 50 -200 nm; between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300 -500 nm; and between 10, 15, 20, 25, 35, 40, 45, 50, 75, 100, 150, 200, 300 -500 nm; and between 10,
- Methods described herein can be used to treat subjects having characteristics or needs defined herein.
- a subject, or a treatment for a subject is selected on the basis of a characteristic described herein.
- the methods described herein allow optimized selection of patients and therapies.
- subjects can be selected or identified prior to subjecting them to any aspects of the methods described herein.
- the subject is selected or is identified as being in need of receiving the AHCM and/or the microenvironment modulator on the basis of optimizing a therapy, e.g., the need for improved delivery and/or efficacy of the therapy (e.g., the cancer therapy).
- a therapy e.g., the need for improved delivery and/or efficacy of the therapy (e.g., the cancer therapy).
- the subject does not have hypertension, or is not being treated for hypertension, at the time of initiation of the AHCM treatment , or at the time of selection of the patient for AHCM administration.
- the subject e.g., patient
- has not been administered a dose of an AHCM e.g., an AHCM named herein, or any AHCM, within 5, 10, 30, 60 or 100 days of, the diagnosis of cancer, or the initiation of the AHCM dosing.
- the subject e.g., a subject with normal or low blood pressure
- the subject is selected or is identified on the basis of being in need of an AHCM and/or the microenvironment modulator, e.g., is selected or is identified as being in need of receiving the AHCM and/or the microenvironment modulator on the basis of optimizing a therapy, e.g., the need for improved delivery and/or efficacy of the therapy (e.g., the cancer therapy).
- a therapy e.g., the need for improved delivery and/or efficacy of the therapy (e.g., the cancer therapy).
- subjects who are in need of receiving the AHCM and/or the microenvironment modulator on the basis of the need for improved delivery or efficacy of the cancer therapy, or optimizing the therapy are the subjects who partially respond or do not respond to the cancer therapy alone.
- an AHCM and/or the microenvironment modulator is selected for treating a subject, on the basis of its ability to optimize a treatment, e.g., a cancer treatment, e.g., improving delivery and/or efficacy of the therapy, e.g., the cancer therapy.
- a treatment e.g., a cancer treatment
- improving delivery and/or efficacy of the therapy e.g., the cancer therapy.
- the subject treated is not a hypertensive patient, e.g., does not have a medical history of high blood pressure, or has not been treated with an antihypertensive agent.
- the subject treated has normal or low mean arterial blood pressure.
- the subject treated has not undergone, or is not being treated with anti-hypertensive therapy.
- the subject has a disorder chosen from one or more of a hyperproliferative disorder, a cancer, a fibrotic disorder, an inflammatory disorder or an autoimmune disorder.
- the subject is in need of cancer therapy.
- the subject is in need of, or being considered for, anti-cancer therapy (e.g., treatment with any of the anti-cancer therapeutics described herein).
- the method includes the step of determining if the subject has a cancer (e.g., a solid or fibrotic cancer), and, responsive to said determination, administering the AHCM and/or the microenvironment modulator, and the anti-cancer agent.
- a cancer e.g., a solid or fibrotic cancer
- the subject is at risk of developing, or having a recurrence of, a cancer, e.g., a subject with pre-neoplasia or a genetic pre-disposition for cancer (e.g., a subject having a BRCAl mutation; or a breast cancer patient treated with in an adjuvant setting (e.g., with tamoxifen)).
- a cancer e.g., a subject with pre-neoplasia or a genetic pre-disposition for cancer (e.g., a subject having a BRCAl mutation; or a breast cancer patient treated with in an adjuvant setting (e.g., with tamoxifen)).
- the subject has early-cancer, or more progressive (e.g., moderate), or metastatic cancer.
- the subject has a solid, fibrotic tumor chosen from one or more of pancreatic (e.g., pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma), breast, colorectal, colon, lung (e.g., small or non-small cell lung cancer), skin, ovarian, prostate, cervix, gastrointestinal (e.g., carcinoid or stromal), stomach, head and neck, kidney, or liver cancer, or a metastatic lesion thereof. Additional examples of cancers treated are described herein below.
- pancreatic e.g., pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma
- lung e.g., small or non-small cell lung cancer
- skin ovarian
- prostate cervix
- gastrointestinal e.g., carcinoid or stromal
- stomach head and neck
- kidney or liver cancer
- the subject has a fibrotic or desmoplastic solid tumor, e.g., a tumor having one or more of: limited tumor perfusion, compressed blood vessels, high interstitial fluid pressure (IFPs), or fibrotic tumor interstitium.
- the subject has a tumor having (e.g., elevated levels of) extracellular matrix components, such as fibers (e.g., collagen, procollagen) and/or polysaccharides (e.g., glycosaminoglycans such as hyaluronan or hyaluronic acid).
- the levels of the extracellular matrix components in the tumor can vary depending on the particular cancer type, the stage of maligancy, and/or in response to cancer therapy. For example, certain tumors may show elevated levels of extracellular matrix components in response to chemotherapy and/or radiation.
- the AHCM alone or in combination with the microenvironment modulator can be administered at any time before, during or after the cancer therapy.
- the subject has a hyperproliferative cancerous condition
- the subject can be one at risk of having the disorder, e.g., a subject having a relative afflicted with the disorder, or a subject having a genetic trait associated with risk for the disorder.
- the subject can be symptomatic or asymptomatic.
- the subject harbors an alteration in an oncogenic gene or gene product.
- the subject is a patient who is undergoing cancer therapy (e.g., the same or other anti-cancer agents, surgery and/or radiation).
- the subject is a patient who has undergone cancer therapy (e.g., other anti-cancer agents, surgery and/or radiation). In one embodiment, the subject has not been treated with the cancer therapy.
- the subject is a patient with a metastatic cancer, e.g., a metastastatic form of a cancer disclosed herein (one or more of pancreatic (e.g., pancreatic adenocarcinoma), breast, colorectal, lung (e.g., small or non-small cell lung cancer), skin, ovarian, or liver cancer.
- a metastatic cancer e.g., a metastastatic form of a cancer disclosed herein (one or more of pancreatic (e.g., pancreatic adenocarcinoma), breast, colorectal, lung (e.g., small or non-small cell lung cancer), skin, ovarian, or liver cancer.
- the subject is a patient having treatment-resistant cancer or hyperproliferative disorder.
- the subject being selected for subjecting to the methods or pharmaceutical compositions herein does not have a renal disease or a disease associated with kidneys.
- the subject treated is a mammal, e.g., a primate, typically a human (e.g., a patient having, or at risk of, a cancer or tumor as described herein).
- a mammal e.g., a primate
- a human e.g., a patient having, or at risk of, a cancer or tumor as described herein.
- the subject treated has a disorder chosen from one or more of a hyperproliferative disorder, a cancer, a fibrotic disorder, an inflammatory disorder or an autoimmune disorder.
- the subject treated has a hyperproliferative disorder, e.g., a hyperpoliferative connective tissue disorder (e.g., a hyperproliferative fibrotic disease).
- a hyperproliferative disorder e.g., a hyperpoliferative connective tissue disorder (e.g., a hyperproliferative fibrotic disease).
- the hyperproliferative fibrotic disease is multisystemic or organ- specific.
- Exemplary hyperproliferative fibrotic diseases include, but are not limited to, multisystemic (e.g., systemic sclerosis, multifocal fibrosclerosis, sclerodermatous graft-versus-host disease in bone marrow transplant recipients, nephrogenic systemic fibrosis, scleroderma), and organ-specific disorders (e.g., fibrosis of the lung, liver, heart, kidney, pancreas, skin and other organs).
- multisystemic e.g., systemic sclerosis, multifocal fibrosclerosis, sclerodermatous graft-versus-host disease in bone marrow transplant recipients, nephrogenic systemic fibrosis, scleroderma
- organ-specific disorders e.g., fibrosis of the lung, liver, heart, kidney, pancreas, skin and other organs.
- the subject treated has a hyperproliferative genetic disorder, e.g., a hyperproliferative genetic disorder chosen from Marfan' s syndrome or Loeys-Dietz syndrome.
- a hyperproliferative genetic disorder chosen from Marfan' s syndrome or Loeys-Dietz syndrome.
- the hyperproliferative disorder e.g., the fibroblast growth factor
- hyperproliferative fibrotic disorder is chosen from one or more of chronic obstructive pulmonary disease, asthma, aortic aneurysm, radiation-induced fibrosis, skeletal-muscle myopathy, diabetic nephropathy, and/or arthritis.
- the AHCM is administered in combination with a microenvironment modulator, and/or a therapy, e.g., a cancer therapy (e.g., one or more of anti-cancer agents, immunotherapy, photodynamic therapy (PDT), surgery and/or radiation).
- a cancer therapy e.g., one or more of anti-cancer agents, immunotherapy, photodynamic therapy (PDT), surgery and/or radiation.
- chemotherapeutic chemotherapeutic agent
- anti-cancer agent are used interchangeably herein.
- the administration of the AHCM and the therapy can be sequential (with or without overlap) or simultaneous.
- Administration of the AHCM and/or the microenvironment modulator can be continuous or intermittent during the course of therapy (e.g., cancer therapy).
- Certain therapies described herein can be used to treat cancers and non-cancerous diseases.
- PDT efficacy can be enhanced in cancerous and non-cancerous conditions (e.g., tuberculosis) using the methods and compositions described herein (reviewed in, e.g., Agostinis, P. et al. (201 1) CA Cancer J. Clin. 61 :250-281).
- administration of the AHCM and/or the microenvironment modulator is initiated prior to the initiation of administration of the therapy (e.g., the cancer therapy), e.g., it is initiated at least one, two, three, or five days, or one, two, three, four, five or more weeks prior to cancer therapy (e.g., the AHCM and/or the microenvironment modulator is administered at a mimimum of two weeks prior to cancer therapy). In an embodiment, it is intiated no more than 5, 10, 20, 30, 60 or 120 days prior to initiation of the therapy, e.g., the cancer therapy.
- administration of the AHCM and/or the microenvironment modulator is initiated prior to the therapy, e.g., the cancer therapy, and the therapy is not initiated until a criterion is met, e.g., a time-based criterion, e.g., administration of AHCM and/or the microenvironment modulator for a predetermined number of days or for a predetermined number of administrations.
- a criterion is met, e.g., a time-based criterion, e.g., administration of AHCM and/or the microenvironment modulator for a predetermined number of days or for a predetermined number of administrations.
- the criterion is meeting a preselected level of AHCM and/or the microenvironment modulator, e.g., a preselected level in serum, plasma or tissue.
- the criterion is meeting a preselected level of a biomarker in plasma, serum or tissue, including but not limited to, an angiotensin receptor (e.g., angiotension-II type-1 receptor; ATi A receptor (ATi A R)), collagen I, collagen III, collagen IV, transforming growth factor beta 1 (TGF- ⁇ ⁇ ), connective tissue growth factor (CTGF), or thrombospondin- 1 (TSP-1).
- an angiotensin receptor e.g., angiotension-II type-1 receptor; ATi A receptor (ATi A R)
- TGF- ⁇ ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- TSP-1 thrombospondin- 1
- microenvironment modulator is sequential and/or concurrent with the therapy, e.g., the cancer therapy, as described herein.
- the AHCM and/or the microenvironment modulator is administered, or a preselected level, e.g., a plasma level, of AHCM and/or the
- microenvironment modulator is maintained for a preselected portion of the time the subject receives the therapy, e.g., the cancer therapy.
- the AHCM and/or the microenvironment modulator therapy is maintained for the entire period in which the therapy, e.g., the cancer therapy, is administered, or for the entire period in which a preselected level of the therapy (e.g., an anti-cancer agent) persists in the subject.
- a preselected level of the therapy e.g., an anti-cancer agent
- therapy with the AHCM and/or the microenvironment modulator continues during the entire therapy, e.g., cancer therapy, schedule.
- administration of the AHCM and/or the microenvironment modulator is discontinued prior to cessation of the therapy, e.g., the cancer therapy.
- administration of the AHCM and/or the microenvironment modulator is continued after cessation of the the therapy, e.g., the cancer therapy, e.g., the administration continues hours, days, months or more, after cessation of the cancer therapy.
- the microenvironment modulator are administered, alone or in combination with the therapy, e.g., the cancer therapy.
- the AHCM is administered at a sub-anti- hypertensive dose and an anti-hypertensive dose during the course of therapy.
- a sub- anti-hypertensive dose of the AHCM can be administered prior to, or at the time, of the therapy, e.g., the cancer therapy (e.g., treatment with an anti-cancer agent that increases mean arterial blood pressure, e.g, treatment with an anti- angiogenic drug (e.g., Avastin, sunitinib or sorafenib)); then followed by a subsequent hypertensive dose of the AHCM.
- the cancer therapy e.g., treatment with an anti-cancer agent that increases mean arterial blood pressure, e.g, treatment with an anti- angiogenic drug (e.g., Avastin, sunitinib or sorafenib)
- the AHCM (alone or in combination) is administered substantially continuously over a period of, or at least 15, 30, 45 minutes; a period of, or at least, 1, 5, 10, 24 hours; a period of, or at least, 2, 5, 10, 14 days; a period of, or at least, 3, 4, 5, 6, 7, 8 weeks; a period of, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 months; a period of, or at least, 1, 2, 3, 4, 5 years, or longer.
- the AHCM is administered as a controlled- or sustained release formulation, dosage form, or device.
- the AHCM is formulated for continuous delivery, e.g., oral, subcutaneus or intravenous continuous delivery.
- the AHCM (alone or in combination with the microenvironment modulator and/or cancer therapy) is in an oral controlled- or extended release dosage form or formulation.
- the AHCM is administered via an implantable device, e.g., a pump (e.g., a subcutaneous pump), an implant or a depot.
- the delivery method can be optimized such that an AHCM dose as described herein (e.g., a standard, sub-hypertensive, or higher than standard dose) is administered and/or maintained in the subject for a pre-determined period (e.g., a period of, or at least: 15, 30, 45 minutes; 1, 5, 10, 24 hours 2, 5, 10, 14 days; 3, 4, 5, 6, 7, 8 weeks; 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 months; 1, 2, 3, 4, 5 years, or longer).
- the substantially continuously or extended release delivery or formulation of the AHCM (with or without the combination of the microenvironment modulator and/or therapy) can be used for prevention or treatment of cancer for a period of hours, days, weeks, months or years.
- the therapy is chosen from one or more of: nanotherapy (e.g., a viral cancer therapeutic agent (e.g., an oncolytic herpes simplex virus (HSV), a lipid nanoparticle (e.g., a liposomal formulation (e.g., pegylated liposomal doxorubicin (DOXIL ® )), or a polymeric nanoparticle); an antibody that binds to a cancer target; an RNAi or antisense RNA agent;, a chemotherapeutic agent (e.g., a cytotoxic or a cytostatic agent); PDT, immunotherapy, radiation; or surgery; or any combination thereof.
- a viral cancer therapeutic agent e.g., an oncolytic herpes simplex virus (HSV), a lipid nanoparticle (e.g., a liposomal formulation (e.g., pegylated liposomal doxorubicin (DOXIL ® )
- the AHCM and the therapy are administered to a subject, e.g., a subject as described herein, in combination with the microenvironment modulator.
- the microenvironment modulator causes one or more of: reduces solid stress (e.g., growth- induced solid stress in tumors); decreases tumor fibrosis; reduces interstitial hypertension or interstitial fluid pressure (IFP); increases interstitial tumor transport; increases tumor or vessel perfusion; increases vascular diameters and/or enlarges compressed or collapsed blood vessels; reduces or depletes one or more of: cancer cells, or stromal cells (e.g., tumor associated fibroblasts or immune cells); decreases the level or production of extracellular matrix components, such as fibers (e.g., collagen, procollagen), and/or polysaccharides (e.g., glycosaminoglycans such as hyaluronan or
- fibers e.g., collagen, procollagen
- polysaccharides e.g.,
- the microenvironment modulator includes an anti- angiogenic therapy, for example, an inhibitor of vascular endothelial growth factor (VEGF) pathway.
- VEGF pathway inhibitors include, but are not limited to, an antibody against VEGF (e.g., bevacizumab); a VEGF receptor inhibitor (e.g., an inhibitor of VEGFR-1 inhibitor, a VEGFR-2 inhibitor, or a VEGFR-3 inhibitor (e.g., VEGFR inhibitors such as Cediranib (AZD2171)); a VEGF trap (e.g., a fusion protein that includes a VEGFR domain (e.g., a VEGFR1 domain 2 and a VEGFR2 domain 3) fused to an Fc fragment of an IgG); and an anti-VEGF aptamer (or a pegylated derivative thereof (e.g., MACUGEN®).
- an antibody against VEGF e.g., bevacizumab
- the microenvironment modulator includes an agent that decreases the level or production of hyaluronic acid, including but not limited to, an antibody against hyaluronic acid, and an anti-hyaluronan enzymatic therapy, such as hyaluronidase or a derivative thereof (e.g., pegylated form thereof) (e.g., PH20, or pegylated, recombinant human hyaluronidase PEGPH20).
- an agent that decreases the level or production of hyaluronic acid including but not limited to, an antibody against hyaluronic acid, and an anti-hyaluronan enzymatic therapy, such as hyaluronidase or a derivative thereof (e.g., pegylated form thereof) (e.g., PH20, or pegylated, recombinant human hyaluronidase PEGPH20).
- the microenvironment modulator includes an inhibitor of the hedgehog pathway, e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58.
- an inhibitor of the hedgehog pathway e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58.
- the microenvironment modulator includes an agent that improves drug penetration in tumors.
- the agent is a disulfide-based cyclic RGD peptide peptide (iRGD) or an analogue thereof.
- the microenvironment modulator includes a taxane therapy apoptosis as described in Griffon-Etienne, G. et al.
- the microenvironment modulator includes an agent that modulates (e.g, inhibits) a hypoxia inducible factor (HIF), for example, an agent that inhibits hypoxia- inducible factors l and 2a (HIF- la and HIF-2a).
- HIF hypoxia inducible factor
- the agent is an antibody against an HIF.
- the agent is an HIF chemical inhibitor, such as phenethyl isothiocyanate (PEITC).
- the microenvironment modulator includes an agent that decreases the level or production of collagen or procollagen.
- an agent that degrades collagen e.g., collagenase.
- the microenvironment modulator is an anti- fibrotic agent or inhibitor of a profibrotic pathway (a "profibrotic pathway inhibitor") (e.g., a pathway dependent- or independent of TGF-beta and/or CTGF activation).
- a profibrotic pathway inhibitor e.g., a pathway dependent- or independent of TGF-beta and/or CTGF activation.
- the AHCM and/or the cancer therapy is administered in combination with one or more of: an inhibitor of endothelin- 1, PDGF, Wnt/beta-catenin, IGF-1, TNF-alpha, and/or IL-4.
- the AHCM and/or the cancer therapy is administered in combination with an inhibitor of endothelin- 1 and/or PDGF.
- the AHCM and/or the cancer therapy is administered in combination with an inhibitor of one or more of: chemokine receptor type 4 (CXCR4) ⁇ e.g., AMD3100, MSX- 122); stromal-derived- factor- l(SDF-l) ⁇ e.g., tannic acid); hedgehog ⁇ e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58).
- CXCR4 chemokine receptor type 4
- SDF-l stromal-derived- factor- l(SDF-l) ⁇ e.g., tannic acid
- hedgehog ⁇ e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58.
- the AHCM and/or the cancer therapy is administered in combination with an anti-fibrotic agent, for example, a pirfenidone (PFD, 5-methyl- l- phenyl-2-(lH)-pyridone), as further described herein.
- an anti-fibrotic agent for example, a pirfenidone (PFD, 5-methyl- l- phenyl-2-(lH)-pyridone), as further described herein.
- the administration of the AHCM, the cancer therapy, the microenvironment modulator and/or the profibrotic pathway inhibitor can be sequential (with or without overlap) or simultaneous ⁇ e.g., a described herein).
- the cancer treated is an epithelial, mesenchymal or hematologic malignancy.
- the cancer treated is a solid tumor ⁇ e.g., carcinoid, carcinoma or sarcoma), a soft tissue tumor ⁇ e.g., a heme malignancy), and a metastatic lesion, e.g., a metastatic lesion of any of the cancers disclosed herein.
- the cancer treated is a fibrotic or desmoplastic solid tumor, e.g., a tumor having one or more of: limited tumor perfusion, compressed blood vessels, high interstitial fluid pressure (IFPs), or fibrotic tumor interstitium.
- IFPs interstitial fluid pressure
- the solid tumor is chosen from one or more of pancreatic ⁇ e.g., pancreatic adenocarcinoma ⁇ e.g., pancreatic ductal adenocarcinoma (PDA)), breast, gastric, colorectal, lung ⁇ e.g., small or non-small cell lung cancer), skin, ovarian, prostate, or liver cancer. Additional examples of cancers treated are described herein below.
- the cancer treated contains ⁇ e.g., has elevated levels of) extracellular matrix components, such as fibers ⁇ e.g., collagen, procollagen) and/or polysaccharides ⁇ e.g., glycosaminoglycans such as hyaluronan or hyaluronic acid).
- extracellular matrix components such as fibers ⁇ e.g., collagen, procollagen) and/or polysaccharides ⁇ e.g., glycosaminoglycans such as hyaluronan or hyaluronic acid.
- the levels of the extracellular matrix components in the cancer can vary depending on the particular cancer type, the stage of maligancy, and/or in response to cancer therapy. For example, certain cancer may show elevated levels of extracellular matrix components in response to chemotherapy and/or radiation.
- the AHCM alone or in combination with the microenvironment modulator can be administered at any time before, during or after the cancer therapy.
- the AHCM and/or the microenvironment modulator is administered in combination with a cancer therapy (e.g., one or more of anti-cancer agents, photodynamic therapy (PDT), immunotherapy, surgery and/or radiation).
- a cancer therapy e.g., one or more of anti-cancer agents, photodynamic therapy (PDT), immunotherapy, surgery and/or radiation.
- the cancer therapy includes one or more of: a cancer therapeutic, including, for example, a nanotherapy (e.g., one or more nanotherapeutic agents, including viral cancer therapeutic agents (e.g., an oncolytic herpes simplex virus (HSV)) a lipid nanoparticle (e.g., a liposomal formulation (e.g., pegylated liposomal doxorubicin (DOXIL ® )), or a polymeric nanoparticle); one or more cancer therapeutic antibodies (e.g., anti-HER2, anti-EGFR, anti- CD20 antibodies); RNAi and antisense RNA agents; one or more chemotherapeutic agents (e.g., low molecular weight chemotherapeutic agents, including a cytotoxic or a cytostatic agent)); photodynamic therapy; immunotherapy; radiation; or surgery, or any combination thereof.
- a nanotherapy e.g., one or more nanotherapeutic agents, including viral cancer therapeutic agents (e.g., an oncolytic
- any combination of one or more AHCMs and one or more therapeutic modalities can be used.
- exemplary cancer therapeutics include, but are not limited to, nanotherapeutic agents (e.g., one or more lipid nanoparticles (e.g., a liposomal formulation (e.g., pegylated liposomal doxorubicin (DOXIL ® ) or liposomal paclitaxel (e.g., Abraxane®)), or a polymeric nanoparticle); one or more low molecular weight chemotherapeutic s (e.g., gemcitabine, cisplatin, epirubicin, 5-fluorouracil, paclitaxel, oxaliplatin, or leucovorin); one or more antibodies against cancer targets (e.g., growth factor receptor such as HER-2/neu, HER3, VEGF)
- nanotherapeutic agents e.g., one or more lipid nanoparticles (e.g., a lip
- the chemotherapeutic agent used in combination with the AHCM and/or the microenvironment modulator is a cytotoxic or a cytostatic agent.
- cytotoxic agents include antimicrotubule agents, topoisomerase inhibitors (e.g., irinotecan), or taxanes (e.g., docetaxel), antimetabolites, mitotic inhibitors, alkylating agents, intercalating agents, agents capable of interfering with a signal transduction pathway, agents that promote apoptosis and radiation.
- the methods can be used in combination with immunodulatory agents, e.g., IL- 1, 2, 4, 6, or 12, or interferon alpha or gamma, or immune cell growth factors such as GM-CSF.
- the cancer therapy includes an immune or
- immunotherapy used in combination with the AHCM, other cancer therapies, and/or the microenvironment modulator, described herein.
- factor such as hypoxia and/or limited perfusion are believed to cause immunosuppression and/or limit the efficacy of certain immune therapies.
- AHCM alone or in combination with therapies described herein, can be used to improve the efficacy of said immune therapies.
- immune therapies include, but are not limited to, CTLA-4 blockade (e.g., an anti-CTLA-4 antibody (e.g., ipilimumab)); immune-based therapies (including, e.g., immune or dendritic cell- based vaccines and antagonists of immune inhibitory signals or
- immune-based therapies include, but are not limited to, e.g., immune or dendritic cell- based vaccines (Seton-Rogers, S. (2012) Nature Reviews Cancer 12:230-231; Palucka, K. et al. (2012) Nature Reviews Cancer 12:265-277); effector memory CD8+ T cells (Bird, L. (2012) Nature Reviews Immunology 12:227);
- the therapy is a cell-based immunotherapy wherein immune cells are expanded ex vivo and injected into the subject.
- the cancer therapy includes PDT used in combination with the AHCM, other cancer therapies, and/or the microenvironment modulator, described herein.
- PDT includes administration of a photosensitizing agent (e.g., a porhyrin, a porpyrin precursor, a chorlin, or a phthalocyanine) followed by irradiation at a wavelength corresponding to an absorbance band of the photosensitizing agent.
- a series of events lead to one or more of: cell death (e.g., tumor cell death), damage to the micro vasculature, or induction of a local inflammatory reaction).
- cell death e.g., tumor cell death
- damage to the micro vasculature e.g., damage to the micro vasculature, or induction of a local inflammatory reaction.
- PDT is reviewed in, e.g., Agostinis, P. et al. (201 1) CA Cancer J. Clin. 61 :250-281.
- the cancer therapy includes an inhibitor of a cancer stem cell (also referred to herein as a "cancer initiating cell”), used in combination with the AHCM, other cancer therapies and/or the microenvironment modulator, described herein.
- a cancer stem cell also referred to herein as a "cancer initiating cell”
- hypoxia and cancer drugs including anti- angiogenic drugs
- radiation therapy are believed to increase the number of cancer stem cells.
- AHCM AHCM
- an inhibitor of a cancer stem cell can be used to reduce the production of these stem cells.
- exemplary inhibitors of cancer stem cells include, but are not limited to, hedgehog (e.g., SMO) antagonists; and Wnt pathway antagonists (e.g., antibody, OMP- 18R5).
- the AHCM and/or the microenvironment modulator, alone or in combination with one or more cancer therapies described herein, are administered for cancer prevention (e.g., alone or in combination with cancer-prevention agents), during periods of active disorder, or during a period of remission or less active disorder.
- the AHCM and/or the microenvironment modulator can be administered for cancer prevention, before treatment or prevention, concurrently with treatment or prevention, post-treatment or prevention, or during remission of the disorder.
- the cancer therapy is administered simultaneously, sequentially, or a combination of both, with the AHCM and/or the microenvironment modulator.
- the AHCM and/or the microenvironment modulator is administered alone or in combination with cancer-prevention agents, e.g., to treat or prevent cancer in high risk subjects (e.g., a subject with pre-neoplasia or a genetic pre-disposition for cancer (e.g., a subject having a BRCA1 mutation); or a breast cancer patient treated with tamoxifen).
- cancer-prevention agents e.g., to treat or prevent cancer in high risk subjects (e.g., a subject with pre-neoplasia or a genetic pre-disposition for cancer (e.g., a subject having a BRCA1 mutation); or a breast cancer patient treated with tamoxifen).
- the AHCM and/or the microenvironment modulator alone or in combination with the cancer therapy, is a first line treatment for the cancer, e.g., it is used in a subject who has not been previously administered another drug intended to treat the cancer.
- the AHCM and/or the microenvironment modulator alone or in combination with the cancer therapy, is a second line treatment for the cancer, e.g., it is used in a subject who has been previously administered another drug intended to treat the cancer.
- the AHCM and/or the microenvironment modulator alone or in combination with the cancer therapy, is a third, fourth, or greater than fourth, line treatment for the cancer, e.g., it is used in a subject who has been previously administered two, three, or more than three, other drugs intended to treat the cancer.
- the AHCM and/or the microenvironment modulator is administered as adjunct therapy, e.g., a treatment in addition to a primary therapy.
- the AHCM and/or the microenvironment modulator is administered as adjuvant therapy.
- the AHCM and/or the microenvironment modulator is administered as neoadjuvant therapy.
- the AHCM and/or the microenvironment modulator is administered to a subject prior to, or following surgical excision/removal of the cancer.
- the AHCM and/or the microenvironment modulator is administered to a subject before, during, and/or after radiation treatment of the cancer.
- the AHCM and/or the microenvironment modulator is administered to a subject, e.g., a cancer patient who will undergo, is undergoing or has undergone cancer therapy (e.g., treatment with a chemotherapeutic agent, radiation therapy and/or surgery).
- a subject e.g., a cancer patient who will undergo, is undergoing or has undergone cancer therapy (e.g., treatment with a chemotherapeutic agent, radiation therapy and/or surgery).
- the AHCM and/or the microenvironment modulator is administered prior to the cancer therapy. In other embodiments, the AHCM and/or the microenvironment modulator is administered concurrently with the cancer therapy. In yet other embodiments, the AHCM and/or the microenvironment modulator is administered prior to the cancer therapy and concurrently with the cancer therapy. In instances of concurrent administration, the AHCM and/or the microenvironment modulator can continue to be administered after the cancer therapy has ceased.
- the AHCM and/or the microenvironment modulator is administered sequentially with the cancer therapy.
- the AHCM and/or the microenvironment modulator can be administered before initiating treatment with, or after ceasing treatment with, the cancer therapy.
- the administration of the AHCM and/or the microenvironment modulator overlaps with the cancer therapy, and continues after the cancer therapy has ceased.
- the AHCM and/or the microenvironment modulator is administered concurrently, sequentially, or as a combination of concurrent administration followed by monotherapy with either the cancer therapy, the AHCM, and/or the microenvironment modulator.
- the method includes administering the AHCM and/or the microenvironment modulator as a first therapeutic agent, followed by administration of a cancer therapy (e.g., treatment with a second therapeutic agent, radiation therapy and/or surgery).
- a cancer therapy e.g., treatment with a second therapeutic agent, radiation therapy and/or surgery
- the method includes administering a cancer therapy first (e.g., treatment with a first therapeutic agent, radiation therapy and/or surgery), followed by administering the AHCM and/or the microenvironment modulator as a second therapeutic agent.
- the method includes administering the AHCM and/or the microenvironment modulator in combination with a second, third or more additional therapeutic agents (e.g., anti-cancer agents as described herein).
- the AHCM and/or the microenvironment modulator and/or the anticancer agent described herein can be administered to the subject systemically (e.g., orally, parenterally, subcutaneously, intravenously, rectally, intramuscularly, intraperitoneally, intranasally, transdermally, or by inhalation or intracavitary installation).
- the AHCMs are administered orally.
- the AHCM and/or the microenvironment modulator and/or the anticancer agent are administered locally or intratumorally (e.g., via an oncolytic virus).
- the AHCM is administered as a pharmaceutical composition comprising one or more AHCMs, and a pharmaceutically acceptable excipient.
- the AHCM is administered, or is present in the
- composition e.g., the pharmaceutical composition (e.g., the same nanoparticle composition).
- the AHCM, the microenvironment modulator and/or the cancer therapy are administered as separate compositions, e.g., pharmaceutical compositions (e.g., nanoparticle compositions).
- the AHCM, the microenvironment modulator, and the cancer therapy are administered separately, but via the same route (e.g., orally or intravenously).
- microenvironment modulator, and the cancer therapy are administered by different routes (e.g., AHCM is administered orally; the microenvironment modulator is administered subcutaneoulsy; and a cancer therapeutic is administered intravenously).
- AHCM is administered orally
- the microenvironment modulator is administered subcutaneoulsy
- a cancer therapeutic is administered intravenously.
- the AHCM, the microenvironment modulator, and the cancer therapy are administered in the same composition, e.g., pharmaceutical composition.
- the methods of the invention can further include the step of evaluating, or monitoring the subject, e.g., for one or more of: tumor size; the level or signaling of one or more of transforming growth factor beta 1 (TGFp i), connective tissue growth factor (CTGF), thrombospondin- 1 (TSP- 1), or an angiotensin receptor (e.g., angiotension-II type-1 receptor; ATIA receptor (ATi A R)); tumor collagen I levels; fibrotic content, interstitial pressure; a plasma, serum or tissue biomarker, e.g., collagen I, collagen III, collagen IV, TGFp i, CTGF, TSP- 1; levels of one or more cancer markers; the rate of appearance of new lesions, metabolism, hypoxia evolution; the appearance of new disease-related symptoms; the size of tissue mass, e.g., a decreased or stabilization; quality of life, e.g., amount of disease associated pain; histological analysis, lobular pattern, and/or the
- the subject can be evaluated or monitored in one or more of the following periods: prior to beginning of treatment; during the treatment; or after one or more elements of the treatment have been administered. Monitoring can be used to evaluate the need for further treatment with the same AHCM, alone or in combination with, the same microenvironment modulator and/or the same anti-cancer agent, or for additional treatment with additional agents. Generally, a decrease in one or more of the parameters described above is indicative of the improved condition of the subject.
- the method includes evaluating (e.g., detecting) the level of an angiotensin receptor (e.g., angiotension-II type-1 receptor; AT1A receptor (AT1AR) in the subject, e.g., in a tumor from the subject. Detection of the angiotensin receptor in the tumor from the subject indicates that the subject is likely to respond to the AHCM.
- angiotensin receptor e.g., angiotension-II type-1 receptor; AT1A receptor (AT1AR)
- the methods of the invention can further include the step of analyzing a nucleic acid or protein from the subject, e.g., analyzing the genotype of the subject.
- the analysis can be used, e.g., to evaluate the suitability of, or to choose between alternative treatments, e.g., a particular dosage, mode of delivery, time of delivery, inclusion of adjunctive therapy, e.g., administration in combination with a second agent, or generally to determine the subject's probable drug response phenotype or genotype.
- the nucleic acid or protein can be analyzed at any stage of treatment, but preferably, prior to administration of the AHCM and/or anti-cancer agent, to thereby determine appropriate dosage(s) and treatment regimen(s) of the AHCM (e.g., amount per treatment or frequency of treatments) for prophylactic or therapeutic treatment of the subject.
- the invention features a pharmaceutically acceptable composition
- a pharmaceutically acceptable composition comprising, in a single dosage form, an AHCM and an anti-cancer agent, e.g., a small molecule or a protein, e.g., an antibody.
- an AHCM and an anti-cancer agent e.g., a small molecule or a protein, e.g., an antibody.
- one or both of the AHCM and the anti-cancer agent are provided in a nanoparticle.
- the AHCM and anti-cancer agent can be in separate or the same entity.
- the AHCM can be provided as a first nanoparticle and the anti-cancer agent provided as a second nanoparticle (e.g., where the second nanoparticle has a structural property (e.g., size or composition) or a functional property (e.g., release kinetics or a pharmacodynamic property) that differs from the first nanoparticle).
- a structural property e.g., size or composition
- a functional property e.g., release kinetics or a pharmacodynamic property
- an AHCM and an anti-cancer agent can be provided on the same entity, e.g., in the same nanoparticle.
- the invention features a pharmaceutically acceptable composition (e.g., nanoparticle) comprising an AHCM, e.g., an AHCM described herein.
- an AHCM e.g., an AHCM described herein.
- the AHCM is in a dosage described herein, e.g., a standard of care dosage form, a sub-anti -hypertensive dosage form, or a greater than a standard of care dosage form.
- the AHCM is formulated in a dosage form that is according to the standard of care anti-hypertensive or anti-heart failure dosage form, e.g., a standard of care dosage form as described herein.
- the AHCM is formulated in a dosage form that is less than the standard of care anti -hypertensive or anti-heart failure dosage form (e.g., a dosage form that is less than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.16, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7-fold, that of the standard of care dosage form, e.g., a standard of care dosage from as described herein).
- the standard of care dosage form e.g., a standard of care dosage from as described herein.
- the AHCM is formulated in a dosage form that is greater than the standard of care anti -hypertensive or anti-heart failure dosage form (e.g., a dosage form that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dosage form, e.g., a standard of care dosage from as described herein).
- the standard of care anti -hypertensive or anti-heart failure dosage form e.g., a dosage form that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dosage form, e.g., a standard of care dosage from as described herein.
- the invention features a pharmaceutically acceptable composition
- a pharmaceutically acceptable composition comprising an anti-cancer agent, e.g., an anti-cancer agent described herein, as a nanoparticle, e.g., a nanoparticle configured for a method described herein.
- the invention features a therapeutic kit that includes the AHCM, alone or in combination with a therapy, e.g., an anti-cancer agent, described herein, and optionally, instructions for use, e.g., for the treatment of cancer.
- a therapy e.g., an anti-cancer agent, described herein
- instructions for use e.g., for the treatment of cancer.
- the kit comprises one or more dosage for or pharmaceutical preparation or nanoparticle described herein
- the invention features a method optimizing access to a target tissue, e.g., a cancer, or optimizing delivery to a target tissue, e.g., a cancer, of an agent, e.g., a systemically administered agent, e.g., a diagnostic or imaging agent.
- the method comprises:
- AHCM collagen modifying agent
- an agent e.g., a diagnostic or imaging agent to said subject.
- the method includes one or more of the following:
- the AHCM is an anti-hypertensive agent and is administered at a standard of care dose, a sub-anti -hypertensive dose, or a greater than a standard of care -anti- dose;
- the agent e.g., diagnostic or imaging agent
- the agent has a hydrodynamic diameter of greater than 1, 5, or 20 nm, e.g., is nanoparticle
- the agent is an imaging agent, e.g., radiologic agent, an NMRA agent, a contrast agent; or
- the subject is treated with a dosing regimen described herein, e.g., AHCM administration is initiated prior to administration of the agent, e.g., for at least one, two, three, or five days, or one, two, three, four, five or more weeks prior to administration of the agent.
- a dosing regimen described herein e.g., AHCM administration is initiated prior to administration of the agent, e.g., for at least one, two, three, or five days, or one, two, three, four, five or more weeks prior to administration of the agent.
- the AHCM is administered in an amount sufficient to alter (e.g., enhance) the distribution or efficacy of the agent. In one embodiment, the AHCM is administered in an amount sufficient to alter (e.g., enhance) the distribution or efficacy of the agent, but in an amount insufficient to inhibit or prevent tumor growth or progression by itself.
- the AHCM is administered at a dose that causes one or more of the following: a decrease in the level or production of collagen, a decrease in tumor fibrosis, an increase in interstitial tumor transport, improvement of tumor perfusion, or enhanced penetration or diffusion, of the cancer therapeutic in a tumor or tumor vasculature, in the subject.
- the subject is further treated with a cancer therapy, e.g., as therapy as described herein.
- the subject is a human, or a non-human animal, e.g., a mouse, a rat, a non-human primate, horse, or cow.
- a non-human animal e.g., a mouse, a rat, a non-human primate, horse, or cow.
- the invention features a diagnostic kit that includes the AHCM, alone or in combination with the agent, e.g., a diagnostic or imaging agent, described herein, and optionally, instructions for use, e.g., for the diagnosis of cancer.
- the agent e.g., a diagnostic or imaging agent, described herein, and optionally, instructions for use, e.g., for the diagnosis of cancer.
- the invention features a method, or assay for, identifying an AHCM.
- the method, or assay includes providing a cancer or a cancer-associated cell (e.g., a culture of a carcinoma associated fibroblast cell); contacting said cancer or a cancer- associated cell with a candidate agent; detecting a change in the cancer cell in the presence, or absence, of the candidate agent.
- the detected change includes one or more of an increase or decrease of TGFp i level, connective tissue growth factor (CTGF) level, or collagen (e.g., collagen 1) level.
- CTGF connective tissue growth factor
- the candidate agent is chosen from one or more of: an antagonist of renin angiotensin aldosterone system (“RAAS antagonist"), an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ATi blocker), a thrombospondin 1 (TSP-1) inhibitor, a transforming growth factor beta 1 (TGF- ⁇ ) inhibitor, and a connective tissue growth factor (CTGF) inhibitor.
- RAAS antagonist an antagonist of renin angiotensin aldosterone system
- ACE angiotensin converting enzyme
- ATi blocker an angiotensin II receptor blocker
- TSP-1 thrombospondin 1
- TGF- ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- the method, or assay can further include the step of comparing the treated methods or assays to a reference value, e.g., a value obtained in the absence of the candidate agent, or by addition of a control agent, e.g., a positive agent (e.g., losartan), or a negative agent (e.g., saline control), and comparing the difference between the treated and control methods.
- a control agent e.g., a positive agent (e.g., losartan), or a negative agent (e.g., saline control
- the method, or assay can be performed in vitro, in vivo, or a combination of both.
- the method, or assay includes: evaluating the candidate agent in vitro, e.g., using a culture of carcinoma associated cells.
- the candidate agent is added to the culture medium; and the condition medium is analyzed for an increase or decrease of TGFp i level, connective tissue growth factor (CTGF) level, or collagen level.
- CTGF connective tissue growth factor
- the candidate agent is administered to a subject, e.g., an animal model, e.g., an animal tumor model.
- the candidate agent is administered to the subject under suitable conditions; and the subject is analyzed for an increase or decrease of TGFpi level, connective tissue growth factor (CTGF) level, or collagen level.
- CTGF connective tissue growth factor
- the levels of these parameters are analyzed as described in the appended Examples.
- candidate agents evaluated using the in vitro assays are tested in vivo.
- the invention features a composition for use, or the use, of a AHCM agent, alone or in combination with an anti-cancer agent described herein for the treatment of a cancer or tumor described herein.
- Headings or numbered or lettered elements e.g., (a), (b), (i) etc, are presented merely for ease of reading.
- the use of headings or numbered or lettered elements in this document does not require the steps or elements be performed in alphabetical order or that the steps or elements are necessarily discrete from one another.
- Fig. 1 is a panel a bar graphs depicting the effects of losartan (10 ⁇ /L) in total and active TGFp levels, and collagen I synthesis by carcinoma associated fibroblasts (CAFs) in vitro.
- Figs. 2A-2B shows the effects of Losartan on collagen production in tumors.
- Fig. 2B shows a dose response curve of the effect of losartan doses of 10, 20 and 60 mg/kg/day in decreasing the SHG levels by 20, 33 and 67%, respectively, at the end of 15 days, indicating a dose-dependent reduction in collagen levels in Losartan-treated tumors.
- Fig. 3 is a bar graph showing a dose response of losartan vs. collagen content in HSTS26T tumors. Losartan treatment at 20 and 60 mg/kg/day led to 42% and 63% reduction in collagen I staining respectively. The staining in each treatment group was compared to a control group that received saline.
- Fig. 4 is a bar graph showing the effect of losartan in decreasing the mean arterial blood pressure (MABP) in mice in a dose-dependent manner.
- MABP mean arterial blood pressure
- Figs. 5A-5D shows the effects of Losartan in collagen levels in tumors.
- Fig. 5A shows the results of Collagen-I and nuclei immuno staining in tumor sections in L3.6pl and MMTV control and losartan (20mg/kg/day) treated tumors.
- Scale bar 100 ⁇ .
- Losartan treatement e.g., at 20 mg/kg/day significantly reduced the collagen levels in the treated tumors.
- Fig. 5B is a bar graph summarizing the effects after two weeks losartan treatment at 20mg/kg/day; losartan treatment significantly reduced the collagen I
- Fig. 5C is a panel of photographs showing collagen-I and nuclei
- Fig. 5D is a bar graph summarizing the effects of Losartan in significantly reducing the collagen-I immunostaining in HSTS26T and Mu89 by 44% (p ⁇ 0.02) and 20% (p ⁇ 0.05), respectively.
- Fig. 6 is a panel of bar graphs showing the effects of losartan on TSP-1, active and total TGF- ⁇ , and collagen I in HSTS26T tumors.
- Treated animals received losartan (15 mg/kg/day) in drinking water. Tumors were excised after two weeks of treatment, homogenized and analyzed for total and activated TGF- ⁇ levels by ELISA. Note a 3.5 fold reduction in TSP-1, a 4 fold reduction in active TGF- ⁇ and a two fold reduction in collagen 1 after losartan treatment (p ⁇ 0.05).
- Fig. 7A shows the effects of losartan in decreasing tumor TSP-1
- Fig. 7B is a bar graph summarizing the effects of losartan treatment in significantly reducing the TSP-1 immununo staining in HSTS26T and MU89 tumors by 73% (p ⁇ 0.04) and 24% (p ⁇ 0.03), respectively.
- Figs. 8A-8C shows the effects of: Losartan in increasing the delivery of nanoparticles and nanotherapeutics.
- Fig. 8A shows two photograhs (control and losartan) and a bar graph summarizing the distribution of intratumorally (i.t.) injected 100 nm diameter nanoparticles in HSTS26T tumors.
- Losartan significantly increased (p ⁇ 0.001) the distribution of i.t- injected nanoparticles in both tumor types (1.5 fold in HSTS26T and 4 fold in Mu89).
- An analysis of the distribution pattern shows control tumors with fewer intratumoral nanoparticles and a majority of nanoparticles that backtracked out of the needle track and accumulated at the tumor surface.
- treated tumors have a significant number of intratumoral nanoparticles.
- Scale bar 100 ⁇ .
- FIG. 8B shows two photograhs (control and losartan) and a bar graph summarizing the distribution of viral infection 24 hrs after the intratumoral injection of HSV expressing the green fluorescent protein.
- HSV infection in control tumors is limited to the cells in close proximity to the injection site whereas losartan treated tumors have a more extensive spread of HSV infection within the tumors.
- Scale bar 1mm. Losartan
- Fig. 8C shows two photograhs (control and losartan) and a bar graph summarizing the distribution of intravenously (i.v.) injected lOOnm diameter nanoparticles in L3.6pl tumors.
- the nanoparticles are localized around perfused vessels.
- Scale bar 100 ⁇ .
- Fig. 9 is a bar graph showing the changes in diffusion coefficient in HSTS26T tumors after losartan treatment.
- the diffusion coefficient of IgG was measured in HSTS26T tumors implanted in the dorsal window chamber of SCID mice. Treated animals received (40 mg/kg/day) losartan by i.p. injection while control animals received saline.
- the results show a significant increase (p ⁇ 0.04) in diffusion coefficient as measured by multiphoton fluorescence recovery after photobleaching (FRAP).
- FRAP multiphoton fluorescence recovery after photobleaching
- Fig. 10 is a representative distribution profile depicting fractions of injected nanospheres present as a function of the distance from a tumor vessel (penetration depth).
- the nanosphere penetration depth was analyzed in frozen sections from tumors resected 24 hrs after the intravenous nanosphere injection.
- the mean characteristic penetration length increased from 18 ⁇ 5 ⁇ (mean ⁇ SE) in control to 37 ⁇ 6 ⁇ in losartan-treated tumors.
- Ten areas per tumor were analyzed in 6 control and 6 treated tumors.
- Fig. 11A-11D shows the effects of Losartan in significantly delaying the growth of tumors treated with DOXIL® or HSV.
- Figs. 11A-11B shows linear graphs of the results from mice bearing HSTS26T (A) and Mu89 (B) tumors treated for 2 weeks with either losartan or saline prior to the i.t. injection of HSV.
- Losartan alone did not affect the growth of Mu89 or HSTS26T tumors.
- the growth delay was significantly longer in HSTS26T tumors treated with losartan and HSV compared to tumors treated with HSV alone.
- the i.t. injection of HSV did not delay the growth of Mu89 tumors, but the combined losartan and HSV treatment significantly retarded the growth of Mu89 tumors.
- Fig. llC shows the effect in tumor volume in mice that received losartan treatment prior to i.v. DOXIL® infusion (losartan and DOXIL®) have smaller tumors than those that received DOXIL® alone (DOXIL® alone) in L3.6pl tumors. Note that there is no difference in tumor size between saline and losartan-treated mice.
- Figs. 12A-12B shows the relationship between the collagen structure and the virus infection and necrosis.
- HSTS26T the dense mesh-like collagen network confined virus infection to the immediate area surrounding the injection point.
- losartan treatment there was a reduction in the density of the network that presumably allowed virus particles to infect a larger area and thus more tumor cells.
- Arrows indicate viable and virus infected cells, respectively.
- Scale bar 10 ⁇ .
- Figs. 13A-13B shows a schematic of virus distribution and infection in Mu89 (A) and HSTS26T (B) tumors.
- the schematics show how the different collagen network structures affect virus propagation and distribution.
- the collagen fibers (1) restrict the movement of virus particles (round spheres, 2) and the infection (3) of non-infected (4) cancer cells.
- Fig. 13A Mu89 tumors, collagen bundles divide the tumor into isolated regions that cannot be traversed by virus particles. Losartan treatment destabilizes the collagen bundles and allows virus particles to move from one region to another.
- the collagen structure is a mesh-like sieve. Virus particles can still propagate through the sieve but do not extend very far from the injection site. Losartan treatment significantly destabilizes the mesh structure in the internal regions of the tumor and allows the virus to propagate and infect a larger area.
- Fig. 14B is a bar graph showing that there is a two-fold increase (p ⁇ 0.05) in necrosis in tumors (both HSTS26T and MU89) that received losartan prior to HSV injection.
- Fig. 15 shows the in vivo proliferation rates for HSTS26T and MU89 after losartan treatment. Tumors were resected and stained for Ki67 to assess proliferation rates. There was no statistically significant difference in positive Ki67 staining after losartan treatment in HSTS26T and MU89 tumors. There was however a significant difference in proliferation between the two tumor types, the number of Ki67 positive cells was 3 fold higher in HSTS26T tumors.
- FIG. 16 shows the results of PCR analysis of AGTR1 expression in CAF, MU89 and HSTS26T cells.
- MU89 cells and CAF express AGTR1 while HSTS26T cells do not.
- HUVECs were used as a positive control.
- GAPDH levels revealed that all three samples had roughly the same amount of cDNA.
- Figs. 17A-17D shows the effects of angiotensin blockade with ATi blockers or ACE inhibitors in normalizing the tumor microenvironment. Studies with an ARB, losartan, are shown.
- Angiotensin blockade (A) diminishes interstitial matrix density in mammary (MMTV) and pancreatic (L3.6PL) tumors in mice, (B) reducing compressive stress in mammary (E0771) and pancreatic (Pan-02) tumors.
- C This increases the fraction of perfused vessels (arrows) in tumors (E0771 shown), resulting in (D) a normalized vascular network (E0771 shown) that is more efficient and effective at drug and oxygen delivery.
- Figs. 18A-18D shows the effects of angiotensin blockade with ATi blockers or ACE inhibitors in improving drug transport and distribution in tumors.
- angiotensin blockade (A) improves tumor oxygenation (E0771 shown) through enhanced perfusion while (B) making blood vessels deliver drugs more rapidly.
- Reorganization of the interstitial matrix also (C) improves penetration of nanoparticles in desmoplastic tumors (L3.6PL shown) (D).
- Figs. 19A-19E shows the effect of angiotensin blockade with ATi blockers or ACE inhibitors in improving the effectiveness of cancer therapy. Studies with an ARB, losartan, are shown.
- Angiotensin blockade, given in combination with chemotherapy improves the effectiveness of the low MW chemotherapeutic doxorubicin in breast cancer models, (B) slowing tumor growth and (C) increasing animal survival (E0771 shown).
- angiotensin blockade improves the effectiveness of the nanoparticle DOXIL® in pancreatic tumors (L3.6PL shown), e.g., by decreasing the tumor weight (D) and/or tumor size or volume (E).
- Figs. 20A-20B shows the compression of tumor blood vessels in human breast cancer. Biopsies of tumors from breast ductal adenocarcinoma patients were stained for CD31 -positive vessels. Unbridled cell proliferation in the confined microenvironment of these tumor and stromal cells results in vessel compression in the stroma (A) and within tumor nodes (B). All vessels appear to be compressed to some degree, with many completely collapsed.
- Figs. 21A-21D are histology images of mouse tumors showing collagen I (blue), CD31-positive vessels (red), and lectin-positive vessels (green), with CD31-lectin co- staining (yellow) denoting perfused vessels. Representative stainings were shown by arrows.
- angiotensin inhibitors improve perfusion of tumor blood vessels.
- Control E0771 breast tumors (A) are dense with collagen I and vessels, yet only a small fraction of these vessels are perfused.
- Control AK4.4 pancreatic tumors C have higher collagen I levels and a lower vessel density, with vessels that are also poorly perfused. Losartan improves perfusion in E0771 (B) and AK4.4 (D) by decreasing collagen I levels, without anti- angiogenic effects.
- Scale bar ⁇ .
- Figs. 22A-22D are bar graphs showing that angiotensin inhibitors improve vascular perfusion.
- FIGs. 22C-22D the CD31 -positive vessel density, measured using histology following angiotensin inhibition using losartan, is shown.
- Losartan does not affect vessel density, as quantified by the vessel number density (C) and the total vessel length (D), indicating no anti-angiogenic effect at this 40mg/kg dose.
- Animal number « 7-9 for all groups.
- Fig. 23 is a bar graph showing that angiotensin inhibitors do not decrease blood pressure at certain doses.
- Losartan and lisinopril treatment at a 40mg/kg dose does not lower blood pressure in these tumor-bearing mice.
- blood pressure in these diseased mice is lower than in healthy FVB mice ( ⁇ 90mmHg).
- Figs. 24A-24D show the effect of angiotensin inhibitors on decompressing vessels by reducing solid stress.
- Figs. 24A-24B the tumor matrix levels following angiotensin inhibition with losartan are shown.
- Fig. 24C are representative histology images of lectin, CD31, and collagen I staining. Representative staining were shown by arrows. A high local collagen I concentration appears to colocalize with collapsed vessels, suggesting that elevated matrix levels in the microenvironment of a tumor vessel directly lead to compression. Scale bar, ⁇ .
- Figs. 26A-26B are bar graphs showing that the ACE-I lisinopril decreases matrix levels and solid stress.
- Figs. 28A-28B are bar graphs showing that angiotensin inhibitors result in a normalized network of perfused vessels.
- Mathematical analysis of perfused vessel network efficiency for delivery was conducted. Perfused vessel networks of E0771 tumors were imaged in three dimensions using multiphoton microscopy pre- and post-treatment (days 2- 5). Analysis of the distance from each point in the tumor to the nearest perfused vessel (A) indicates that losartan decreases the maximum distance drugs and oxygen must travel to reach tumor cells.
- Figs. 29A-29E are graphs showing that angiotensin inhibitors increase drug and oxygen delivery.
- Fig. 29A shows small-molecule drug delivery to tumors and various organs after angiotensin inhibition with losartan.
- Figs. 29B-29C show oxygen delivery to tumors measured by phosphorescence quenching microscopy during angiotensin inhibition using losartan.
- Losartan increases oxygenation in some tumors (C), whereas all control tumors decrease in oxygen levels. Losartan also appears to result in a more homogenous distribution of well-oxygenated tumor tissue.
- Scale bar 100 ⁇ .
- Fig. 29E shows the penetration rates for nanoparticles after angiotensin inhibition with losartan.
- Penetration rates are quantified as effective permeability, which is the transvascular mass flux per unit vascular surface area and transvascular concentration difference. Closed symbols (top) denote averages by mouse, while open symbols (bottom) are individual tumors.
- Figs. 30A-30B are histology images of mouse tumors showing that angiotensin inhibitors reduce hypoxia. Pimonidazole hypoxia staining (blue), CD31 -positive vessels (red), and lectin-positive vessels (green) are shown, with CD31 -lectin co-staining (yellow) denoting perfused vessels. Representative stainings were shown by arrows. Control E0771 breast tumors (A) show pronounced hypoxia away from the few vessels that are perfused. Losartan improves perfusion, reducing hypoxia (B). Scale bar, ⁇ .
- Fig. 32 is a line graph showing that increasing fluid flow in tumors can improve nanoparticle penetration. Predictions of physiologically-based mathematical model of how modulating interstitial hydraulic conductivity can improve nanoparticle penetration are shown. Increasing interstitial hydraulic conductivity results in more rapid penetration rates (effective permeability) for all sizes of nanoparticles by allowing for more rapid fluid flow driven by the difference in the microvascular pressure and the interstitial fluid pressure at the tumor margin.
- Fig. 33 is a bar graph showing that angiotensin inhibitors synergistically enhance chemotherapy effectiveness.
- Volumes of orthotopic AK4.4 pancreatic tumors on day 7 in response to treatment with losartan or saline control (40mg/kg daily from day 0-7) in combination with either the small-molecule chemotherapeutic 5-FU or saline control (60mg/kg on days 2 and 6) are shown.
- Statistical tests were corrected for multiple comparisons using the Holm-Bonferroni method.
- Fig. 34 is a survival curve showing that angiotensin inhibitors do not decrease survival.
- Animal survival following tumor implantation, with initiation of treatment with losartan on day 11, is whon.
- Losartan monotherapy does not affect survival versus saline.
- Animal number n 5-6.
- Figs. 35A-35C are graphs showing that angiotensin inhibitors enchance chemotherapy in multiple models.
- Fig. 35A the volumes of orthotopic 4T1 breast tumors in response to treatment with losartan or saline control (40mg/kg daily from day 0 on) in combination with either the small-molecule chemotherapeutic doxorubicin or saline control (2mg/kg every 3 days from day 1 on) are shown.
- Fig. 35B the quantification of tumor growth rates, based on the time to reach double the initial volume, is shown.
- Fig. 35C the animal survival following the initiation of treatment is shown.
- Animal number n 6-7 for all groups. Statistical tests were corrected for multiple comparisons using the Holm-Bonferroni method.
- Figs. 36A-36C show the effects of vascular normalization using anti- angiogenic therapy on nanoparticle delivery in tumors
- Fig. 36A nanoparticle penetration versus particle size in orthotopic 4T1 mammary tumors in response to normalizing anti- angiogenic therapy with the VEGF receptor inhibitor, DC101. Nanoparticle concentrations are relative to initial intravascular levels, with vessels in black. Normalization improves 12nm particle penetration, while not detactably affecting 125nm penetration. Scale bar, ⁇ .
- FIG. 37A-37C are representative images of immunohistochemiccal staining for collagen I in AK4.4 tumor samples from mice administered with vehicle (PBS) or losartan.
- mice were subcutaneously injected with PBS (Group 1 or Gl).
- mice were administered with losartan via subcutaneous pump (Group 2 or G2).
- mice were subcutaneously injected with losartan in the absence of pump (Group 3 or G3).
- the percentages of collagen I positive areas are 22.356%, 4.453%, and 1 1.34% for Groups 1-3, respectively.
- Fig. 38A is a bar graph showing the average percentages of collagen I positive areas in tumor samples from mice subcutaneously injected with PBS (Group 1), administered with losartan via subcutaneous pump (Group 2), and subcutaneously injected with losartan in the absence of pump.
- the average percentages of collagen I positive areas are 16.26 ⁇ 1.72%, 3.24 ⁇ 0.48%, and 8.71 ⁇ 0.65% for Groups 1-3, respectively.
- Fig. 38B is a bar graph showing the average numbers of collagen I fibers in tumor samples from mice subcutaneously injected with PBS (Group 1), administered with losartan via subcutaneous pump (Group 2), and subcutaneously injected with losartan in the absence of pump.
- the average numbers of collagen I positive fibers are 28.04 ⁇ 2.41, 10.01 ⁇ 1.28, and 17.93 ⁇ 1.14 for Groups 1-3, respectively.
- the invention is based, at least in part, on the discovery that anti-hypertensive and/or collagen-modifying (AHCM) agents (including angiotensin inhibitors, e.g., angiotensin receptor blockers (e.g., losartan) and angiotensin-converting enzyme inhibitors (ACE-I) improve the delivery and efficacy of cancer therapeutics.
- AHCM anti-hypertensive and/or collagen-modifying
- AHCM anti-hypertensive and/or collagen-modifying agents
- angiotensin inhibitors e.g., angiotensin receptor blockers (e.g., losartan)
- ACE-I angiotensin-converting enzyme inhibitors
- the abnormal matrix of tumors limits the delivery of nano-therapeutics in many types of cancer, e.g., pancreatic, breast, lung, colorectal.
- the overgrowth of fibrous tissue impedes the movement of nanotherapeutics in tumors two mechanisms - viscoelastic and steric hindrances.
- Fibrous tissue is highly viscoelastic, meaning it is quite thick and stiff, and therefore slows the movement of these drugs to a small fraction of their typical speed.
- This tissue is basically an extremely dense mesh, with small pores that are about the same size as nanotherapeutics, thus it does not allow much space for these drugs and often halts their movements by confining them close to blood vessels (in case of intravenous injection) or near the site of injection (in case of intra-tumor injection).
- This barrier is found in all solid tumors, with possible exception of brain tumors, though it is most prominent in pancreatic, breast, lung, and colorectal cancers.
- Nanotherapeutics owing to their large size relative to the pores that form the tumor microenvironment, are especially hindered by fibrous tissue.
- losartan prevents the production of matrix molecules like collagen, which are a component of the dense mesh of fibrous tissue.
- losartan is believed to act on fibroblasts and tumor cells by inhibiting the TGF-beta and CTGF pathways, thus limiting their pro-fibrotic activity. It does so by blocking the activity of the angiotensin-II type- 1 receptor (ATI), which is highly expressed on both fibroblasts and tumor cells in a variety of cancers.
- ATI angiotensin-II type- 1 receptor
- losartan blocks activity downstream of ATI in various signaling pathways, including the activation of TGF-beta and CTGF. Since these two pathways promote the production of collagen and other components of fibrotic tissue, blocking them will allow the fibrosis to subside. The result is tissue that is much more like the normal surrounding organ, and is therefore easier to penetrate.
- nanotherapeutics in tumors allowing them to penetrate tumors more easily, and allows these drugs to distribute more widely throughout tumors, making them more effective at fighting tumor growth.
- losartan makes nanotherapeutics more effective against cancer.
- losartan normalizes the collagen, interstitial matrix of several solid tumors, thus facilitating the penetration of
- chemotherapeutics such as large molecular weight (e.g., nano-) chemotherapeutics.
- losartan reduced collagen I levels in carcinoma associated fibroblasts (CAFs) isolated from breast cancer biopsies, and caused a dose-dependent reduction in stromal collagen in desmoplastic models of human breast, pancreatic and skin tumors in mice.
- CAFs carcinoma associated fibroblasts
- Losartan also improved the distribution, therapeutic efficacy and/or penetration of nanopartices (e.g., oncolytic herpes simplex viruses (HSV) and pegylated liposomal doxorubicin (DOXIL ® )).
- nanopartices e.g., oncolytic herpes simplex viruses (HSV) and pegylated liposomal doxorubicin (DOXIL ® )
- nanotherapeutics are not as limited by the interstitial matrix barriers, but are similarly affected by other barriers such as abnormal and collapsed blood vessels.
- losartan is shown to facilitate decompression of blood vessels, thus improving tumor perfusion and delivery of low molecular weight
- chemotherapeutics thus facilitating radiation and chemotherapeutic delivery through vascular normalization.
- these agents improve delivery of molecules as small as oxygen - a radiation and chemo sensitizer - through vascular normalization (Figs. 18A-18B), while also enhancing the penetration of larger agents through interstitial matrix normalization (Fig. 18C, 18D).
- these agents enhance the effectiveness of low molecular weight chemotherapeutics, as well as nanotherapeutics in breast and pancreatic cancer models - leading to reduced tumor growth and longer animal survival (Figs. 19A-19E).
- angiotensin inhibitors other than losartan
- angiotensin inhibitors including, for example, angiotensin receptor blockers (ARBs), such as candesartan and valsartan, as well as angiotensin converting enzyme inhibitors (ACE-I), such as lisinopril (see e.g., Figs. 26-27).
- ARBs angiotensin receptor blockers
- ACE-I angiotensin converting enzyme inhibitors
- lisinopril see e.g., Figs. 26-27.
- angiotensin inhibitors e.g., angiotensin receptor blockers
- ACE inhibitors can enhance the delivery of a therapy, and thus have broad applicability for combination therapy with all classes of anti-cancer agents, including low molecular weight, small-molecule chemotherapeutics, biologies, nucleic acid agents and nanoparticle therapies.
- the AHCM described herein e.g., angiotensin inhibitors, such as angiotensin receptor blockers and ACE inhibitors
- a microenvironment modulator to enhance penetration and/or diffusion, of a cancer therapy in a tumor or tumor vasculature, in a subject.
- Such combination may cause one or more of: reduce solid stress (e.g., growth-induced solid stress in tumors); decrease tumor fibrosis; reduce interstitial hypertension or interstitial fluid pressure (IFP); increase interstitial tumor transport; increase tumor or vessel perfusion; increase vascular diameters and/or enlarge compressed or collapsed blood vessels; reduce or deplete one or more of: cancer cells, or stromal cells (e.g., tumor associated fibroblasts or immune cells); decrease the level or production of extracellular matrix components, such as fibers (e.g., collagen, procollagen), and/or polysaccharides (e.g., glycosaminoglycans such as hyaluronan or hyaluronic acid); decrease the level or production of collagen or procollagen; decreases the level or production of hyaluronic acid; increases tumor oxygenation; decreases tumor hypoxia; decreases tumor acidosis; enable immune cell infiltration; decreases immunosuppression; increases antitumor immunity; or decreases cancer stem
- Exemplary microenvironment modulators include, but are not limited to, an anti-angiogenic therapy, for example, an inhibitor of vascular endothelial growth factor (VEGF) pathway; an agent that decreases the level or production of hyaluronic acid; an inhibitor of the hedgehog pathway; an agent that improves drug penetration in tumors (e.g., a disulfide-based cyclic RGD peptide peptide (iRGD) or an analogue thereof); a taxane therapy (e.g., taxane-induced apoptosis); an agent that decreases the level or production of collagen or procollagen; and/or a profibrotic pathway inhibitor as described herein.
- an anti-angiogenic therapy for example, an inhibitor of vascular endothelial growth factor (VEGF) pathway; an agent that decreases the level or production of hyaluronic acid; an inhibitor of the hedgehog pathway; an agent that improves drug penetration in tumors (e.g., a disulfide-based cycl
- Angiotensin blockers offer numerous advantages over other approaches, including anti-angiogenic therapies, anti-collagen agents and other matrix modifiers.
- anti-angiogenic therapies normalize the vasculature alone and have been approved for only a limited number of indications.
- ARBs and ACE-Is are FDA-approved as anti-hypertensives with manageable adverse effects.
- anti-angiogenics which are FDA-approved adjuncts that enhance drug delivery to tumors, tend not to improve the delivery for larger particles as they can reduce the size of "pores" in vessel walls.
- vascular normalization with anti-angiogenic therapies can typically enhance the delivery and effectiveness of small therapeutics, including small molecule chemotherapeutics, biologies and small nanoparticles (e.g., in the range of 1- 12 nm), while not substantially affecting the delivery of larger therapeutics (e.g., about 50 nm to about 100 nm, e.g., about 60 nm or larger) (data shown in Figs. 36A-36C).
- anti-collagen agents such as relaxin, can improve transport through the tumor matrix, but not facilitate the delivery of low molecular weight agents (see e.g., US 6,719,977).
- AHCM e.g., angiotensin blockers
- chemotherapeutics chemotherapeutics, biologies, nucleic acid agents and nanoparticle therapies (as described in the Examples herein).
- Matrix modifiers like bacterial collagenase, relaxin, and matrix
- metalloproteinase-1 and -8 have been used to modify the collagen or proteoglycan network in tumors and have improved the efficacy of intratumorally (i.t.) injected oncolytic viruses (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 J-H, et al. (2006) J Natl Cancer Inst 98: 1482-1493).
- these agents may produce normal tissue toxicity (e.g., bacterial collagenase) or increase the risk of tumor or metastatic progression (e.g., relaxin, matrix metalloproteinases).
- matrix- degrading enzymes which can normalize the collagen matrix, are not selective for tumors and can increase invasion and metastasis.
- Other approaches for improving interstitial transport may also cause increased metastasis.
- relaxin a hormone produced during pregnancy that modulates collagen fiber structure to improve diffusion of nano-sized probes (Brown, E. et al. (2003) Nat Med. 9(6):796-800; Perentes, J.Y. et al. (2009) Nat. Methods 6(2): 143-5), may lead to increased metastasis, perhaps due to the mechanism of relaxin as a matrix-degrading therapy.
- ARBs and ACE-Is have no significant complications associated with matrix remodeling in normal tissues, leading to their safety as anti-hypertensives.
- losartan monotherapy did not significantly increase metastasis in the cancer model tested, AK4.4; and losartan combination with 5-FU appeared to reduce the incidence and size of metastases (Table 2).
- angiotensin inhibitors such as ARBs and ACE-Is, are likely to cause less metastasis than other anti-collagen agents, such as matrix- degrading enzymes and relaxin.
- ARBs and ACE-Is as small-molecule agents, are that they can also be delivered via nanovectors containing chemotherapeutics (e.g., liposomes, nano-particles) to enhance their localization to tumors to further limit toxicity.
- chemotherapeutics e.g., liposomes, nano-particles
- compositions for treating or preventing a cancer e.g., a solid tumor such as a desmoplastic tumor
- a cancer e.g., a solid tumor such as a desmoplastic tumor
- an anti-hypertensive agent as a single agent or combination with a microenvironment modulator and/or a cancer therapeutic agent (for example, a therapeutic agent ranging in size from a large nanotherapeutic to a low molecular weight chemotherapeutics and/or oxygen) are disclosed.
- Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5%, 4%, 3%, 2% or 1% of a given value or range of values.
- Delivery refers to the placement of the agent(s) in sufficient proximity to one or more (or all) of: the tumor vasculature, the tumor interstitial matrix, or tumor cells or tumor-associated cells (e.g., fibroblasts), to have a desired effect.
- the agent(s) can be, e.g., a cancer therapy (e.g., a cancer therapeutic agent(s) as described herein), or a diagnostic or imaging agent(s).
- agent or “agent(s)” as used generically herein can include one, two or more agents.
- the cancer therapeutic agent includes, e.g., one or more of a small molecule, a protein or a nucleic acid drug, an oncolytic virus, a vaccine, an antibody or a fragment thereof, or a combination thereof.
- the cancer therapeutic agent can be "free" or packaged or formulated into a delivery vehicle, e.g., a particle, e.g., a nanoparticle (e.g., a lipid nanoparticle, a polymeric nanoparticle, or a viral particle). Delivery of a therapeutic agent is characterized by placement of the therapeutic agent in sufficient proximity to the cell to alter an activity of the cell, e.g., to kill the cell and/or reduce its ability to divide.
- the agent is a diagnostic or an imaging agent (e.g., one or more of a radiologic agent, an NMRA agent, a contrast agent, or the like).
- the diagnostic or imaging agent can be "free” or packaged or formulated into a delivery vehicle. Delivery of a diagnostic or imaging agent is characterized by placement of the agent in sufficient proximity to a target cell or tissue to allow detection of the target cell or tissue.
- increased (or improved) delivery (as compared with a delivery which is the same or similar except that it is carried out in the absence of an AHCM) can include one or more of: increased delivery to, or amount or concentration in, the tumor vasculature, of the agent;
- the tumor e.g., the tumor vasculature interstitial matrix, of the agent
- tumor cells or tumor-associated cells e.g., fibroblasts
- the agent in the tumor e.g., the tumor interstitial matrix
- non-tumor tissue e.g., peripheral blood
- TGF-beta pathway inhibition of the TGF-beta pathway in the tumor, e.g., in the tumor vasculature interstitial matrix;
- the tumor e.g., the tumor vasculature interstitial matrix
- an extracellular matrix component such as a fiber (e.g., collagen, procollagen), and/or a polysaccharide (e.g., a glycosaminoglycan such as hyaluronan or hyaluronic acid);
- a fiber e.g., collagen, procollagen
- a polysaccharide e.g., a glycosaminoglycan such as hyaluronan or hyaluronic acid
- hyaluronan levels in the tumor e.g., the tumor vasculature interstitial or stromal matrix.
- increased (or improved) delivery (as compared with a delivery which is the same or similar except that it is carried out in the absence of an AHCM) can also include increased amount of the agent distributed to at least a portion of the tumor.
- the increased amount of the agent delivered to the tumor in the presence of the AHCM can be distributed homogenously or heterogeneously throughout the tumor.
- Effectiveness as used herein in the context of therapy, e.g., cancer therapy, can be characterizes as the extent to which a therapy has a desired effect, including but not limited to, alleviation of a symptom, diminishment of extent of disease, stabilized state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
- Improved efficacy in the context of efficacy of cancer therapy, can be characterized by one or more of the following: an increase in an anti -tumor effect, of the cancer therapy, and/or a lessening of unwanted side effects (e.g., toxicity), of the cancer therapy, as compared with a treatment which is the same or similar except that it is carried out in the absence of treatment with an AHCM.
- the increase in the antitumor effect of the cancer therapy includes one or more of: inhibiting primary or metastatic tumor growth; reducing primary or metastatic tumor mass or volume; reducing size or number of metastatic lesions; inhibiting the development of new metastatic lesions; reducing one or more of non- invasive tumor volume or metabolism; providing prolonged survival; providing prolonged progression-free survival; providing prolonged time to progression; and/or enhanced quality of life.
- the term "improved efficacy" as used herein, with respect to a cancer therapy in combination with an AHCM can refer to an increase in reduction of primary or metastatic tumor growth by 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%, up to and including 100%, as compared to the reduction of primary or metastatic tumor growth during a cancer therapy alone (i.e., in the absence of an AHCM).
- the administration of an AHCM in combination with a cancer therapy can increase the reduction of primary or metastatic tumor growth by 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 higher, as compared to the reduction of primary or metastatic tumor growth during a cancer therapy alone (i.e., in the absence of an AHCM).
- Methods for monitoring tumor growth in vivo are well known in the art, e.g., but not limited to, X-ray, CT scan, MRI and other art-recognized medical imaging methods.
- the term "improved efficacy" as used herein, with respect to a cancer therapy in combination with an AHCM can refer to an increase in perfusion of an anti-cancer agent (e.g., low molecular weight therapeutics or nanotherapeutics such as DOXIL® or immune cells) into a tumor, e.g., by 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%, up to and including 100%, as compared to perfusion of an anti-cancer agent alone (i.e., in the absence of an AHCM).
- combination with a cancer therapy can increase perfusion of an anti-cancer agent (e.g., low molecular weight therapeutics or nanotherapeutics such as DOXIL®) into a tumor, by 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 higher, as compared to the perfusion efficiency of an anti-cancer agent alone (i.e., in the absence of an AHCM).
- an anti-cancer agent e.g., low molecular weight therapeutics or nanotherapeutics such as DOXIL®
- Methods to measure tumor perfusion in vivo are well established in the art, including, but not limited to, positron emission tomography (PET), and ultrasound or contrast-enhanced ultrasound.
- the term "improved efficacy" as used herein, with respect to a cancer therapy in combination with an AHCM can refer to an increase in reduction in expression level of at least one biomarker, e.g., at least one cancer biomarker (e.g., in a biological sample such as a blood sample, a serum sample, a plasma sample or a tissue biopsy), e.g., by 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%, up to and including 100%, as compared to the reduction in expression level of the at least one cancer biomarker when administered with a cancer therapy, alone (i.e., in the absence of an AHCM).
- at least one cancer biomarker e.g., in a biological sample such as a blood sample, a serum sample, a plasma sample or a tissue biopsy
- the administration of an AHCM in combination with a cancer therapy can increase the reduction in expression level of at least one biomarker (e.g., in a biological sample such as a blood sample, a serum sample, a plasma sample or a tissue biopsy) by 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 higher, as compared to the reduction in expression level of the at least one cancer biomarker when administered with a cancer therapy alone (i.e., in the absence of an AHCM).
- at least one biomarker e.g., in a biological sample such as a blood sample, a serum sample, a plasma sample or a tissue biopsy
- Examples of a biomarker in the serum, plasma or tissue can include, but are not limited to, TGF-beta 1, TGF-beta 2, CTGF, TSP- 1, collagen I, collagen II, collagen III, or collagen IV.
- Expression levels of biomarkers can be measured on a transcript level and/or a protein level, using any art-recognized analytical methods, e.g., PCR, western blot, ELISA, and/or immunostaining.
- “Blood pressure” is usually classified based on the systolic and diastolic blood pressures.
- “Systolic blood pressure” or Psys refers to the blood pressure in vessels during a heart beat.
- “Diastolic blood pressure” or Pdias refers to the pressure between heartbeats.
- a systolic or the diastolic blood pressure measurement higher than the accepted normal values for the age of the individual is classified as prehypertension or hypertension.
- a systolic or the diastolic blood pressure measurement lower than the accepted normal values for the age of the individual is classified as hypotension.
- a "normal" systolic pressure for an adult is typically in the range of 90-120 mmHg; a “normal” diastolic pressure is usually in the range of 60-80 mmHg.
- the average blood pressure can range from 110/65 to 140/90 mmHg for an adult; 95/65 mmHg for a 1 year infant, and 100/65 mmHg for a 6-9 year old.
- Hypertension has several subclassifications including, prehypertension (120/80 to 139/89 mmHg); hypertension stage I (140/90 to 159 to 99 mmHg), hypertension stage II (greater or equal to 160/100 mmHg, and isolated systolic hypertension (greater or equal to 140/90 mmHg).
- Isolated systolic hypertension refers to elevated systolic pressure with normal diastolic pressure and is common in the elderly. These classifications are made after averaging a patient's resting blood pressure readings taken on two or more office visits.
- Hypertension is generally diagnosed on the basis of a persistently high blood pressure. Usually this requires three separate sphygmomanometer measurements at least one week apart. Often, this entails three separate visits to the physician's office. Initial assessment of the hypertensive patient should include a complete medical history and physical examination.
- hypertension refers to a prehypertensive or a hypertensive stage having a systolic pressure of 120 or greater
- MAP mean arterial pressure
- CO cardiac output
- SVR systemic vascular resistance
- CVP central venous pressure
- MAP (CO x SVR) + CVP.
- MAP can be approximately determined from measurements of the systolic pressure (Psys) and the diastolic pressure (Pdias), while there is a normal resting heart rate, MAP is approximately Pdias + l/3(Psys -Pdias).
- Anti-hypertensive agent refers to an agent (e.g., a compound, a protein) that when administered at a selected dose (referred to herein as “an antihypertensive dose”) reduces blood pressure, typically in a patient (e.g., a hypertensive patient).
- an antihypertensive dose typically reduces blood pressure, typically in a patient (e.g., a hypertensive patient).
- Anti-hypertensive agents are routinely used clinically to treat patients with high blood pressure at doses known in the art.
- Exemplary anti-hypertensive agents include but are not limited to, renin angiotensin aldosterone system antagonists ("RAAS antagonists"), angiotensin converting enzyme (ACE) inhibitors, and angiotensin II receptor blockers (ATi blockers).
- RAAS antagonists renin angiotensin aldosterone system antagonists
- ACE angiotensin converting enzyme
- ATi blockers
- Sub-anti-hypertensive dose refers to a dose of an anti- hypertension agent that is typically less than the lowest dose that would be used to treat a patient for high blood pressure.
- a sub-anti-hypertensive dose has one or more of the following properties:
- blood pressure e.g., the mean arterial blood pressure
- the subject e.g., a hypertensive subject
- the ability of a dose to meet one or more of these standards can be made as measured after a preselected number of dosages, e.g., 1, 2, 5, or 10, or after sufficient dosages that a steady state level, e.g., plasma level, is attained.
- AHCM can be an agent having one or more of the following properties:
- RAAS antagonist renin angiotensin aldosterone system
- ACE angiotensin converting enzyme
- angiotensin II receptor blocker (ATi blocker)
- TSP-1 thrombospondin 1
- TGF- ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- Treating typically refers to one or more of the following: inhibiting primary or metastatic tumor growth;
- AHCM Agents Anti-Hypertensive and/or Collagen Modifying Agents
- the AHCM agent used in the methods and compositions of the invention can be chosen from one or more of: an antagonist of renin angiotensin aldosterone system ("RAAS antagonist"), an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ATi blocker), a thrombospondin 1 (TSP- 1) inhibitor, a transforming growth factor beta 1 (TGF- ⁇ ) inhibitor, and a connective tissue growth factor (CTGF) inhibitor.
- RAAS antagonist an antagonist of renin angiotensin aldosterone system
- ACE angiotensin converting enzyme
- ATi blocker an angiotensin II receptor blocker
- TSP-1 thrombospondin 1
- TGF- ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- renin angiotensin aldosterone system examples include, but are not limited to, aliskiren (TEKTURNA®, RASILEZ®), remikiren (Ro 42- 5892), enalkiren (A-64662), SPP635, and derivatives thereof.
- ACE inhibitors include, but are not limited to, benazepril (LOTENSIN®), captopril (CAPOTEN®), enalapril (VASOTEC®), fosinopril (MONOPRIL®), lisinopril (PRTNIVIL®, ZESTRIL®), moexipril (UNIVASC®), perindopril (ACEON®), quinapril (ACCUPRIL®), ramipril (ALTACE®), trandolapril (MAVIK®), and derivatives thereof.
- angiotensin II receptor blockers include, but are not limited to, losartan (COZAAR®), candesartan (ATACAND®), eprosartan mesylate
- TEVETEN® EXP 3174
- irbesartan AZAPRO®
- L158,809 L158,809
- olmesartan BENICAR®
- saralasin a segment of telmisartin
- MICARDIS® a segment of telmisartin
- DIOVAN® a segment of telmisartin
- the ATi blocker is losartan, or a derivative thereof.
- Losartan is an anti -hypertensive agent with miminal safety risks (Johnston CI (1995) Lancet 346: 1403-1407). Furthermore, in addition to its antihypertensive properties, losartan is also an antifibrotic agent that has been shown to reduce the incidence of cardiac and renal fibrosis (Habashi JP, et al. (2006) Science 312: 117-121; and. Cohn RD, et al. (2007) Nat Med 13:204-210).
- TGF- ⁇ active transforming growth factor- ⁇
- AGTR1 angiotensin II type I receptor
- TGF- ⁇ activators like thrombospondin-1 (TSP-1)
- Habashi JP et al. (2006) Science 312: 1 17-121
- Cohn RD et al. (2007) Nat Med 13:204- 210
- Lavoie P et al. (2005) JHypertens 23 : 1895-1903
- Chamberlain JS (2007) Nai ed 13 : 125-126; and Dietz HC (2010) J Clin Invest 120:403 ⁇ 107.
- Exemplary thrombospondin 1 (TSP-1) inhibitors include, but are not limited to, ABT-510, CVX-045, LSKL, and derivatives thereof.
- Exemplary transforming growth factor beta 1 (TGF- ⁇ ) inhibitors include, but are not limited to, CAT- 192, fresolimumab (GC1008), LY 2157299, Peptide 144 (P144), SB- 431542, SD-208, compounds described in U.S. Patent Serial No. 7,846,908 and U.S. Patent Application Publication No. 2011/0008364, and derivatives thereof.
- CTGF connective tissue growth factor
- Exemplary beta-blockers include, but are not limited to, atenolol
- BLOCADREN® BLOCADREN®
- acebutolol SECTRAL®
- penbutolol LVATOL®
- pindolol pindolol
- carvedilol COREG®
- labetalol NORMODYNE®, TRANDATE®
- the AHCM agent is a TGF- ⁇ inhibitor, e.g., an anti- TGF- ⁇ antibody, a TGF- ⁇ peptide inhibitor.
- the TGF- ⁇ inhibitor is chosen from one or more of: CAT-192, fresolimumab (GC1008), LY 2157299, Peptide 144 (P144), SB-431542, SD-208, compounds described in U.S. Patent Serial No. 7,846,908 and U.S. Patent Application Publication No. 201 1/0008364, or a derivative thereof.
- Suitable doses for administration of the AHCM agent can be evaluated based on the standard of care anti-hypertensive doses of the AHCM agents are available in the art.
- Exemplary standard of care anti-hypertensive and anti-heart failure doses and dosage formulations for ATi inhibitors in humans are as follows: 25- 100 mg day "1 of losartan (available in a dosage form for oral administration containing 12.5 mg, 25 mg, 50 mg or 100 mg of losartan); 4 to 32 mg day "1 of candesartan (ATACAND®) ⁇ e.g., available in a dosage form for oral administration containing 4 mg, 8 mg, 16 mg, or 32 mg of candesartan); 400 to 800 mg day "1 of eprosartan mesylate (TEVETEN®) ⁇ e.g., available in a dosage form for oral administration containing 400 or 600 mg of eprosartan); 150 to 300 mg day "1 of irbesartan (AVAPRO®) ⁇ e.g., available in a dosage form for oral administration containing 150 or 300 mg of irbesartan); 20 to 40 mg day "1 of olmesartan (
- Exemplary standard of care anti-hypertensive and anti-heart failure doses and dosage formulations for ACE inhibitors in humans are as follows: 10 to 40 mg day “1 of benazepril (LOTENSIN®) (Lotensin (benazepril) is supplied as tablets containing 5 mg, 10 mg, 20 mg, or 40 mg of benazepril hydrochloride for oral administration); 25 to 100 mg day "1 of captopril (CAPOTEN®) (available in a dosage form for oral administration containing 12.5 mg, 25 mg, 50 mg or 100 mg of captopril); 5 to 40 mg day "1 of enalapril (VASOTEC®) (available in a dosage form for oral administration containing 2.5 mg, 5 mg, 10 mg or 20 mg of enalapril; 10 to 40 mg day "1 of fosinopril (MONOPRIL®) (available in a dosage form for oral administration containing 10 mg, 20 mg, or 40 mg of fosinopril
- the AHCM agent is administered at a standard of care anti-hypertensive and anti-heart failure doses and dosage formulations, e.g., a dose or dosage formulation as described herein.
- a sub-anti-hypertensive dose or dosage formulation of the AHCM agent is desirable, e.g., a dose of the AHCM agent that is less than the standard of care dose or dosage formulation.
- the sub-anti-hypertensive dose or dosage formulation has a minimal effect in blood pressure in a hypertensive subject (e.g., decreases the mean arterial blood pressure in a hypertensive subject by less than 20%, 10%, or 5% or less).
- the AHCM agent is administered at a dose or dosage formulation that is less than 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, that of the standard of care anti-hypertensive dose (e.g., the lower standard of care dose).
- the dose or dosage formulation is in the range of, for example, 0.01-0.9-fold, 0.02-0.8-fold, 0.05-0.7-fold, 0.1-0.5 fold, 0.1-0.2-fold, that of the standard of care dose or dosage formulation for anti -hypertensive or anti-heart failure use.
- Standard of care doses or dosage formulation of the AHCM are available in the art, some of which are exemplified herein.
- the AHCM agent is administered at a dose or dosage formulation that is greater than the standard of care dose or dosage formulation for anti-hypertensive or anti-heart failure use (e.g., a dose or dosage form that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dose for anti-hypertensive or anti-heart failure use).
- the dose or dosage formulation is in the range of, for example, 1.1 to 10-fold, 1.5-5-fold, 1.7 to 4-fold, or 2-3-fold, that of the standard of care dose or dosage formulation for anti-hypertensive or anti-heart failure use.
- Standard of care doses or dosage formulation of the AHCM are available in the art, some of which are exemplified herein.
- the standard of care dose and dosage forms are provided herein for a number of AHCMs, e.g., losartan.
- the dose and/or dosage form is less than (or higher than) the standard of care dose and/or dosage form. In an exemplary embodiment, it is less than 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 0.7, 0.8, 0.9-fold, that of the standard of care dose or dosage form.
- the dose or dosage form contains an amount of AHCM that is within a range of the reduced amounts of the standard of care dose and/or dosage form.
- an AHCM dosage form that is 0.01-0.9-fold, 0.02-0.8-fold, 0.05-0.7-fold, 0.1-0.5 fold, 0.1- 0.2-fold, that of the standard of care dose or dosage form.
- the range of the dose or the dosage form is 0.5-2.0 times a reduced dose or dosage form recited herein, so long as the dose or dosage form value is less than the standard of care dose or dosage form.
- a standard of care dosage form for losartan is 12.5 mg.
- the dosage form is 0.125 mg (.01x12.5 mg); 0.625 mg (0.05 x 12.5 mg); 1.25 mg (0.1 x 12.5 mg); 2.5 mg (0.2 x 12.5 mg); or 6.25 mg (0.5 x 12.5 mg).
- This calculation can be applied to any standard of care dose and/or dosage form for any AHCM described herein.
- the value is less than the standard of care values. In other embodiments, the value is greater than the standard of care values.
- the dose of the AHCM agent is calculated based on the severity of the fibrosis in the tumor sample.
- the dose of the AHCM agent can be a sub-anti- hypertensive dose, which does not have any anti-tumor effect, e.g., no significant effect on inhibiting or preventing tumor growth or progression when administered alone.
- the dose of the AHCM agent can be comparable to or greater than the standard of care dose or dosage formulation for anti-hypertensive or anti-heart failure use, and does not have any anti-tumor effect, e.g., no significant effect on inhibiting orpreventingng tumor growth or progression when administered alone.
- the invention relates to a method of treating a disorder, e.g., a hyperproliferative disorder (e.g., a cancer) by administering to a patient an AHCM agent, alone or in combination with a therapy or a therapeutic agent, e.g., an anti-cancer agent as described herein.
- a disorder e.g., a hyperproliferative disorder (e.g., a cancer)
- a therapy or a therapeutic agent e.g., an anti-cancer agent as described herein.
- the terms “treat,” “treating” and “treatment” refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as the development or spread of cancer.
- Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
- “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.
- Those in need of treatment include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.
- therapeutic treatment can refer to inhibiting or reducing tumor growth or progression after administration in accordance with the methods or administration with the pharmaceutical compositions described herein.
- 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.
- tumor growth or progression is inhibited or reduced by more than 50%, e.g., at least about 60%, or at least about 70%, after treatment.
- tumor growth or progression is inhibited or reduced by at least about 80%, at least about 90% or greater, as compared to a control (e.g. in the absence of the pharmaceutical composition described herein).
- the therapeutic treatment refers to alleviation of at least one symptom associated with cancer.
- Measurable lessening includes any statistically significant decline in a measurable marker or symptom, such as measuring a cancer biomarker, such as serum/plasma cancer biomarker in a blood sample, after treatment.
- at least one cancer biomarker or sympton 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%.
- at least one cancer biomarker or sympton is alleviated by more than 50%, e.g., at least about 60%, or at least about 70%.
- at least one cancer biomarker or sympton is alleviated by at least about 80%, at least about 90% or greater, as compared to a control (e.g. in the absence of the pharmaceutical composition described herein).
- the terms “prevent,” “preventing” and “prevention” contemplate an action that occurs before a patient begins to suffer from the regrowth of the cancer and/or which inhibits or reduces the severity of the cancer.
- a "therapeutically effective amount" of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of the disorder (e.g., cancer), or to delay or minimize one or more symptoms associated with the disorder (e.g., cancer).
- a therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapeutic agents, which provides a therapeutic benefit in the treatment or management of the disorder.
- the term "therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of the disorder (e.g., cancer), or enhances the therapeutic efficacy of another therapeutic agent.
- a prophylactically effective amount of a compound is an amount sufficient to prevent a disorder (e.g., regrowth of the cancer, or one or more symptoms associated with the cancer, or prevent its recurrence).
- a prophylactically effective amount of a compound means an amount of the compound, alone or in combination with other therapeutic agents, which provides a prophylactic benefit in the prevention of the disorder.
- the term “prophylactically effective amount” can encompass an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent.
- the term "patient” or “subject” refers to an animal, typically a human (i.e., a male or female of any age group, e.g., a pediatric patient (e.g, infant, child, adolescent) or adult patient (e.g., young adult, middle-aged adult or senior adult) or other mammal, such as a primate (e.g., cynomolgus monkey, rhesus monkey); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs; and/or birds, including commercially relevant birds such as chickens, ducks, geese, and/or turkeys, that will be or has been the object of treatment, observation, and/or experiment.
- a human i.e., a male or female of any age group, e.g., a pediatric patient (e.g, infant, child, adolescent) or adult patient (e.g., young adult, middle-aged adult or
- compositions described herein can also be used to treat domesticated animals or pets such as cats and dogs.
- cancer and “tumor” are synonymous terms.
- cancer therapy and “cancer treatment” are synonymous terms.
- chemotherapy As used herein, “chemotherapy,” “chemotherapeutic,” “chemotherapeutic agent” and “anti-cancer agent” are synonymous terms.
- the AHCM agent alone or in combination, is a first line treatment for the cancer, i.e., it is used in a subject who has not been previously administered another drug intended to treat the cancer.
- the AHCM agent alone or in combination, is a second line treatment for the cancer, i.e., it is used in a subject who has been previously administered another drug intended to treat the cancer.
- the AHCM agent alone or in combination, is a third or fourth line treatment for the cancer, i.e., it is used in a subject who has been previously administered two or three other drugs intended to treat the cancer.
- the AHCM agent is administered to a subject before, during, and/or after radiation or surgical treatment of the cancer.
- the AHCM agent is administered, alone or in combination with a cancer therapy or an anti-cancer agent, to a subject who previously did not respond to at least one cancer therapy or anti-cancer agent, including at least two, at least three, or at least four cancer therapies or anti-cancer agents.
- the AHCM agent can be administered to a subject in combination with the cancer therapy or anticancer agent to which he/she previously did not respond, or in combination with a cancer therapy or anti-cancer agent different from the one(s) he/she has been treated with.
- the AHCM agent is administered as adjunct therapy, i.e., a treatment in addition to primary therapy.
- the adjuvant effect of the AHCM administered in combination with a primary therapy can be additive.
- the AHCM alone or in combination with a microenvironment modulator and/or a therapy or a therapeutic agent, e.g., an anti-cancer agent as described herein can be used to treat or prevent a disorder, e.g., a hyperproliferative disorder (e.g., a cancer).
- a disorder e.g., a hyperproliferative disorder (e.g., a cancer).
- the disorder is chosen from one or more of a hyperproliferative disorder, a cancer, a fibrotic disorder, an iinflammatory disorder or an autoimmune disorder.
- the cancer is an epithelial, mesenchymal or hematologic malignancy.
- the cancer treated is a solid tumor (e.g., carcinoid, carcinoma or sarcoma), a soft tissue tumor (e.g., a heme malignancy), and a metastatic lesion, e.g., a metastatic lesion of any of the cancers disclosed herein.
- the cancer treated is a fibrotic or desmoplastic solid tumor, e.g., a tumor having one or more of: limited tumor perfusion, compressed blood vessels, fibrotic tumor interstitium, or increased interstitial fluid pressure.
- the solid tumor is chosen from one or more of pancreatic (e.g., pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma), breast, colon, colorectal, lung (e.g., small cell lung cancer (SCLC) or non- small cell lung cancer (NSCLC)), skin, ovarian, liver cancer, esophageal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney, or prostate cancer.
- pancreatic e.g., pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma
- lung e.g., small cell lung cancer (SCLC) or non- small cell lung cancer (NSCLC)
- SCLC small cell lung cancer
- NSCLC non- small cell lung cancer
- hyperproliferative cancerous disease or disorder all neoplastic cell growth and proliferation, whether malignant or benign, including all transformed cells and tissues and all cancerous cells and tissues.
- Hyperproliferative diseases or disorders include, but are not limited to, precancerous lesions, abnormal cell growths, benign tumors, malignant tumors, and "cancer.”
- cancer refers to an abnormal mass of tissue that results from excessive cell division, in certain cases tissue comprising cells which express, over-express, or abnormally express a hyperproliferative cell protein.
- a cancer, tumor or tumor tissue comprises “tumor cells” which are neoplastic cells with abnormal growth properties and no useful bodily function. Cancers, tumors, tumor tissue and tumor cells may be benign or malignant. A cancer, tumor or tumor tissue may also comprise "tumor-associated non-tumor cells", e.g., vascular cells which form blood vessels to supply the tumor or tumor tissue. Non-tumor cells may be induced to replicate and develop by tumor cells, for example, the induction of angiogenesis in a tumor or tumor tissue.
- cancers include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers are noted below and include: squamous cell cancer (e.g.
- lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, as well as head and neck cancer.
- lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer
- cancer includes primary malignant cells or tumors (e.g., those whose cells have not migrated to sites in the subject's body other than the site of the original malignancy or tumor) and secondary malignant cells or tumors (e.g., those arising from metastasis, the migration of malignant cells or tumor cells to secondary sites that are different from the site of the original tumor).
- primary malignant cells or tumors e.g., those whose cells have not migrated to sites in the subject's body other than the site of the original malignancy or tumor
- secondary malignant cells or tumors e.g., those arising from metastasis, the migration of malignant cells or tumor cells to secondary sites that are different from the site of the original tumor.
- cancers or malignancies include, but are not limited to: Acute Childhood Lymphoblastic Leukemia, Acute Lymphoblastic Leukemia, Acute
- Lymphocytic Leukemia Acute Myeloid Leukemia, Adrenocortical Carcinoma, Adult (Primary) Hepatocellular Cancer, Adult (Primary) Liver Cancer, Adult Acute Lymphocytic Leukemia, Adult Acute Myeloid Leukemia, Adult Hodgkin's Disease, Adult Hodgkin's Lymphoma, Adult Lymphocytic Leukemia, Adult Non-Hodgkin's Lymphoma, Adult Primary Liver Cancer, Adult Soft Tissue Sarcoma, AIDS-Related Lymphoma, AIDS-Related Malignancies, Anal Cancer, Astrocytoma, Bile Duct Cancer, Bladder Cancer, Bone Cancer, Brain Stem Glioma, Brain Tumors, Breast Cancer, Cancer of the Renal Pelvis and Ureter, Central Nervous System (Primary) Lymphoma, Central Nervous System Lymphoma, Cerebellar Astrocytoma, Cerebral Astrocytoma, Cervical Cancer, Childhood
- Neoplasm/Multiple Myeloma Primary Central Nervous System Lymphoma, Primary Liver Cancer, Prostate Cancer, Rectal Cancer, Renal Cell Cancer, Renal Pelvis and Ureter Cancer, Retinoblastoma, Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoidosis Sarcomas, Sezary Syndrome, Skin Cancer, Small Cell Lung Cancer, Small Intestine Cancer, Soft Tissue Sarcoma, Squamous Neck Cancer, Stomach Cancer, Supratentorial Primitive
- Neuroectodermal and Pineal Tumors T-Cell Lymphoma, Testicular Cancer, Thymoma, Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvis and Ureter, Transitional Renal Pelvis and Ureter Cancer, Trophoblastic Tumors, Ureter and Renal Pelvis Cell Cancer, Urethral Cancer, Uterine Cancer, Uterine Sarcoma, Vaginal Cancer, Visual Pathway and Hypothalamic Glioma, Vulvar Cancer, Waldenstrom's Macroglobulinemia, Wilms' Tumor, and any other hyperproliferative disease, besides neoplasia, located in an organ system listed above.
- the AHCM agent as described above and herein, is used to treat a hyperproliferative disorder, e.g., a hyperpoliferative connective tissue disorder (e.g., a hyperproliferative fibrotic disease).
- a hyperproliferative disorder e.g., a hyperpoliferative connective tissue disorder (e.g., a hyperproliferative fibrotic disease).
- the hyperproliferative fibrotic disease is multisystemic or organ-specific.
- Exemplary hyperproliferative fibrotic diseases include, but are not limited to, multisystemic (e.g., systemic sclerosis, multifocal fibrosclerosis, sclerodermatous graft- versus-host disease in bone marrow transplant recipients, nephrogenic systemic fibrosis, scleroderma), and organ-specific disorders (e.g., fibrosis of the eye, lung, liver, heart, kidney, pancreas, skin and other organs).
- the disorder is chosen from liver cirrhosis or tuberculosis.
- the subject treated has a hyperproliferative genetic disorder, e.g., a hyperproliferative genetic disorder chosen from Marfan' s syndrome or Loeys-Dietz syndrome.
- a hyperproliferative genetic disorder chosen from Marfan' s syndrome or Loeys-Dietz syndrome.
- Losartan has been shown to treat human Marfan syndrome, a connective tissue disorder caused by mutations in the gene that encodes the extracellular matrix protein, fibrillin- 1 (Dietz, H.C. et al. (2010) New Engl J Med 363(9) 852-863).
- Fibrillin- 1 comprises the microfibrils of elastic tissue and a component of many other connective tissues.
- Affected patients with Marfan syndrome have blood vessel abnormalities such as aortic aneurysms.
- the vascular disease can result in blood vessel rupture and death in childhood and later in life.
- Dietz et al. first found in mouse models of Marfan syndrome that excessive activation of latent TGF- ⁇ has an important role in the pathophysiology. They used losartan in the affected mice and showed striking effects in improving blood vessel architecture and prevented the development of aortic aneurysms. They have also used losartan to treat children with Marfan syndrome and demonstrated that the drug can strikingly prevent progression of aortic and muscular lesions.
- Aortic diseases other than Marfan syndrome can also benefit from the use of losartan.
- Inhibition of activation of latent TGF- ⁇ locally and decreasing circulating levels of active TGF- ⁇ thus can have effects on components of connective tissues other than collagen in the extracellular matrix of cancer tissues that alter delivery and efficacy of nanotherapeutics.
- the hyperproliferative disorder e.g., the fibroblast growth factor
- hyperproliferative fibrotic disorder is chosen from one or more of chronic obstructive pulmonary disease, asthma, aortic aneurysm, radiation-induced fibrosis, skeletal-muscle myopathy, diabetic nephropathy, and/or arthritis.
- the disorder is chosen from an inflammatory or an autoimmune disorder chosen from multiple sclerosis, inflammatory bowel disease, scleroderma, lupus, rheumatoid arthritis or osteoarthritis.
- the inflammatory disorder is an inflammatory disorder of: the gastrointestinal tract or a gastrointestinal organ, e.g., colitis, Crohn's disease, inflammatory bowel disease (IBD), Barrett's esophagus and chronic gastritis; the lung (e.g., asthma, chronic obstructive pulmonary disease (COPD); the skin (e.g., psoriasis), the cardiovascular system (e.g., atherosclerosis, cholesterol metabolic disorders, oxygen free radical injury, ischemia), the nervous system (e.g., Alzheimer's disease, multiple sclerosis), liver (e.g., hepatitis), kidney (e.g., nephritis), and the pancreas (e.g., pancreatitis).
- colitis Crohn's disease
- IBD inflammatory bowel disease
- COPD chronic obstructive pulmonary disease
- the skin e.g., psoriasis
- the cardiovascular system e.g., athe
- the inflammatory disorder is associated with an autoimmune disorder, e.g., arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis, lupus-associated arthritis, autoimmune thyroiditis or ankylosing spondylitis); scleroderma; lupus; systemic lupus erythematosis; HIV; Sjogren's syndrome; vasculitis; multiple sclerosis; dermatitis (including atopic dermatitis and eczematous dermatitis), myasthenia gravis, inflammatory bowel disease (IBD), Crohn's disease, colitis, diabetes mellitus (type I); acute inflammatory conditions (e.g., endotoxemia, sepsis and septicaemia, toxic shock syndrome and infectious disease); transplant rejection and allergy.
- arthritis including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, p
- the AHCM agent as described above and herein, can be administered in combination with one or more additional therapies, e.g., such as radiation therapy, PDT, surgery, immune therapy, and/or in combination with one or more therapeutic agents, to treat the cancers described herein.
- additional therapies e.g., such as radiation therapy, PDT, surgery, immune therapy, and/or in combination with one or more therapeutic agents, to treat the cancers described herein.
- compositions can be administered concurrently with, prior to, or subsequent to, one or more other additional therapies or therapeutic agents.
- each agent will be administered at a dose and/or on a time schedule determined for that agent.
- the additional therapeutic agent utilized in this combination can be administered together in a single composition or administered separately in different compositions.
- the particular combination to employ in a regimen will take into account compatibility of the inventive pharmaceutical composition with the additional therapeutically active agent and/or the desired therapeutic effect to be achieved.
- the AHCM and/or the therapy is administered in combination with a microenvironment modulator.
- the combined administration of the AHCM and the microenvironment modulator can be used to enhance the efficacy (e.g., penetration and/or diffusion), of a therapy, e.g., a cancer therapy, in a tumor or tumor vasculature in a subject.
- Such combination may cause one or more of: reduce solid stress (e.g., growth-induced solid stress in tumors); decrease tumor fibrosis; reduce interstitial hypertension or interstitial fluid pressure (IFP); increase interstitial tumor transport; increase tumor or vessel perfusion; increase vascular diameters and/or enlarge compressed or collapsed blood vessels; reduce or deplete one or more of: cancer cells, or stromal cells (e.g., tumor associated fibroblasts or immune cells); decrease the level or production of extracellular matrix components, such as fibers (e.g., collagen, procollagen), and/or polysaccharides (e.g., glycosaminoglycans such as hyaluronan or hyaluronic acid); decrease the level or production of collagen or procollagen; decreases the level or production of hyaluronic acid; increases tumor oxygenation; decreases tumor hypoxia; decreases tumor acidosis; enables immune cell infiltration; decreases
- reduce solid stress e.g., growth-induced solid stress
- cancer stem cells also referred to herein as tumor initiating cells
- Exemplary microenvironment modulators include, but are not limited to, an anti-angiogenic therapy, for example, an inhibitor of vascular endothelial growth factor (VEGF) pathway; an agent that decreases the level or production of hyaluronic acid; an inhibitor of the hedgehog pathway; an agent that improves drug penetration in tumors.
- the agent is a disulfide-based cyclic RGD peptide peptide (iRGD) or an analogue thereof; a taxane therapy (e.g., taxane-induced apoptosis); an agent that decreases the level or production of collagen or procollagen; an anti-fibrotic agent and/or a profibrotic pathway inhibitor.
- the microenvironment modulator includes an anti- angiogenic therapy, for example, an inhibitor of vascular endothelial growth factor (VEGF) pathway.
- VEGF pathway inhibitors include, but are not limited to, an antibody against VEGF (e.g., bevacizumab); a VEGF receptor inhibitor (e.g., an inhibitor of VEGFR-1 inhibitor, a VEGFR-2 inhibitor, or a VEGFR-3 inhibitor (e.g., VEGFR inhibitors such as Cediranib (AZD2171)); a VEGF trap (e.g., a fusion protein that includes a VEGFR domain (e.g., a VEGFR1 domain 2 and a VEGFR2 domain 3) fused to an Fc fragment of an IgG); and an anti-VEGF aptamer (or a pegylated derivative thereof (e.g., MACUGEN®).
- an antibody against VEGF e.g., bevacizumab
- the microenvironment modulator includes an agent that decreases the level or production of hyaluronic acid (HA).
- HA hyaluronic acid
- PEGPH20 pegylated derivative of hyaluronidase
- PDA pancreatic ductal adenocarcinoma
- hyaluronidase derivatives in combination with standard chemotherapeutic agents (e.g., gemcitabine), can remodel the tumor microenvironment and increase overall survival (see e.g., Provenzano, P. et al. (2012) Cancer Cell 21 : 418-429).
- AHCM and the microenvironment modulator can be used to enhance penetration and/or diffusion of a cancer therapy in a tumor or tumor vasculature, by for example, decreasing certain matrix components, e.g., HA, in the stroma.
- HA-depleting agents include, but are not limited to, an anti-hyaluronan enzymatic therapy such as hyaluronidase or a derivative thereof (e.g., pegylated recombinant human hyaluronidase) (e.g., PH20, PEGPH20); and an antibody against hyaluronic acid.
- the microenvironment modulator includes an inhibitor of the hedgehog pathway.
- Hedgehog inhibitors have been shown to increase vessel density in pancreatic tumors (Olive, K.P. et al. (2009) Science 324: 1457-61), presumably by reducing stromal cell density and solid stress.
- Exemplary hedgehog inhibitors include, but are not limited to, IPI-926, GDC-0449, cylopamine or an analogue thereof, and GANT58.
- the microenvironment modulator includes an agent that improves drug penetration in tumors.
- the agent is a disulfide-based cyclic RGD peptide peptide (iRGD) or an analogue thereof (e.g., described in Sugahara, KN et al. (2010) Science 328: 1031-5; Ye, Y. et al. (2011) Bioorg Med Chem Lett. 21(4): 1146- 50).
- the microenvironment modulator includes a taxane therapy apoptosis as described in Griffon-Etienne, G. et al.
- the microenvironment modulator includes an agent that modulates (e.g, inhibits) a hypoxia inducible factor (HIF), for example, an agent that inhibits hypoxia- inducible factors l and 2a (HIF- la and HIF-2a).
- HIF activity has been shown to be involved in inflammation (e.g., rheumatoid arthritis) and angiogenesis associated with cancer tumor growth.
- HIF inhibitors such as phenethyl isothiocyanate (PEITC) are under investigation for anti-cancer effects (Syed Alwi SS, et al. (2010) Br. J. Nutr. 104 (9): 1288-96; Semenza GL (2007). Drug Discov.
- the agent is an antibody against an HIF.
- the agent is an HIF chemical inhibitor, such as phenethyl isothiocyanate (PEITC).
- the microenvironment modulator includes an agent that decreases the level or production of collagen or procollagen.
- an agent that degrades collagen e.g., collagenase.
- the AHCM and/or the therapy is administered in combination with a microenvironment modulator chosen from an anti-fibrotic agent or an inhibitor of a profibrotic pathway (a "profibrotic pathway inhibitor") (e.g., a pathway dependent- or independent of TGF-beta and/or CTGF activation).
- a profibrotic pathway inhibitor e.g., a pathway dependent- or independent of TGF-beta and/or CTGF activation.
- the AHCM and/or the cancer therapy is administered in combination with one or more of: an inhibitor of endothelin- 1, PDGF, Wnt/beta-catenin, IGF-1, TNF-alpha, and/or IL-4.
- the AHCM and/or the cancer therapy is administered in combination with an inhibitor of endothelin- 1 and/or PDGF.
- the AHCM and/or the cancer therapy is administered in combination with an inhibitor of one or more of chemokine receptor type 4 (CXCR4) (e.g., AMD3100, MSX- 122); stromal-derived-factor-l(SDF-l) (e.g., tannic acid); hedgehog (e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58).
- CXCR4 chemokine receptor type 4
- SDF-l e.g., tannic acid
- hedgehog e.g., IPI-926, GDC-0449, cylopamine or an analogue thereof, or GANT58.
- an inhibitor of a CXCR4 receptor and/or its ligand, SDF- 1 is administered in combination with a therapy (e.g., a cancer or hyperproliferative therapy as described herein).
- a therapy e.g., a cancer or hyperproliferative therapy as described herein.
- Certain embodiments may further include administration of a further AHCM and/or a microenvironment modulator as described herein.
- inhibition of CXCR4 receptor and/or its ligand, SDF- 1, alone or in combination with an AHCM, e.g., an angiotensin II receptor blocker can be used to reduce the desmoplasia in certain fibrotic or desmoplastic cancers, e.g., a fibrotic or a desmoplastic solid tumor, such as pancreatic cancers (e.g., pancreatic ductal adenocarcinoma (PDAC)).
- a fibrotic or desmoplastic cancers e.g., a fibrotic or a desmoplastic solid tumor, such as pancreatic cancers (e.g., pancreatic ductal adenocarcinoma (PDAC)
- PDAC pancreatic ductal adenocarcinoma
- SDF-1 a/CXCR4 and angiotensin II (ATII) signaling pathways is known to promote carcinoma activated fibroblasts (CAF) recruitment, activation, and
- ATII signaling can stimulate CAF proliferation (Hama, K. et a;. (2006) Biochemical and Biophysical Research Communications, 340: 742-750; Hama, K. et al. (2004) Biochem Biophys Res Commun. 315: 905-911; Shimizu, K. et al. (2008) J Gastroenterol Hepatol, 23 Suppl 1: S I 19- 121), and ATII signaling through ATII-receptor type 1 (ATI) can stimulate CAF matrix production via TGF- ⁇ and ERK-dependent mechanisms (Rodriguez- Vita, J. et al. (2005) Circulation 111: 2509-2517; Yang, F. et al.
- ATII also induces TGF- ⁇ (Elenbaas, B. and Weinberg, R. A. (2001) Experimental Cell Research, 264: 169- 184) and SDF- la (Chu, P. Y. et al. (2010) Am J Pathol, 176: 1735-1742) expression by both cancer cells and CAFs, which can promote CAF proliferation and matrix production.
- TGF- ⁇ Elenbaas, B. and Weinberg, R. A. (2001) Experimental Cell Research, 264: 169- 184)
- SDF- la Cho, P. Y. et al. (2010) Am J Pathol, 176: 1735-1742
- SDF-1 inhibition of a CXCR4 receptor and/or its ligand, SDF-1, can be used (alone or with an inhibitor of ATII signaling) to enhance the distribution of a therapy in fibrotic or
- Exemplary SDF- 1/CXCR4 inhibitors that can be used include, but are not limited to, 2,2'-bicyclam; 6,6'- bicyclam; AMD3100 (IUPAC name: l,l'-[l,4-phenylene- bis(methylene)]-bis-l, 4,8,1 1 -tetraazacyclotetradecane), as described in e.g., U.S. Pat. Nos.
- RNA inhibitors e.g., antisense, siRNAs
- Plerixafor trade name: Mozobil; IUPAC name: 1, 1 '- [1,4-Phenylenebis(methylene)]bis [1,4,8, 1 1 -tetraazacyclotetradecane
- CXCR4 peptide inhibitors or analogs e.g., T-140 analogs (e.g., 4F-benzoyi-TNI4003, TC 14012, TE1401 1, TC14003), CTCE-0214; CTCE-9908; and CP-1221, as well as other inhibitors such as antibodies against SDF-1 or CXCR4, RNA inhibitors (e.g., antisense, siRNAs), among others.
- Exemplary inhibitors are described in, for example, Tamamura, H. et al. Org. Biomol. Chem. 1 :3656-3662, 2003; FEB S Letter 550: 1 -3 (2003 ): 79-83; Wong, D. et al. (2008) Clin. Cancer Res. 14(24): 7975-7980; US Patent Publications 2010/0055088; 2009/0221683;
- the AHCM and/or the cancer therapy is administered in combination with an anti-fibrotic agent, for example, a pirfenidone.
- Pirfenidone PFD or 5-methyl- l-phenyl-2(lH)-pyridone,commercially available from Marnac, Inc.
- Pirfenidone has been shown to produce anti-fibrotic effects in several organs such as the heart, liver, lung and kidney.
- PFD has been shown to have an inhibitory effect on fibroblast growth and collagen synthesis by reducing expression of profibrotic cytokines such as TGF-b (Iyer, S.N. et al.
- PFD has also been shown to reduce leiomyoma cell proliferation and collagen production in cultured cells, as well as reduce TGF-b expression in human malignant glioma cells ⁇ see e.g., Byung-Seok, L. et al. (1998) J of Clinical Endocrinology and Metabolism 83(1): 219-223; and Burghardt, I. et al. (2007) Biochem and Biophys Res. Comm. 354:542-547).
- the AHCM and/or the microenvironment modulator is administered in combination with a low or small molecular weight chemotherapeutic agent.
- exemplary low or small molecular weight chemotherapeutic agents include, but not limited to, 13-cis-retinoic acid (isotretinoin, ACCUTANE®), 2-CdA (2-chlorodeoxyadenosine, cladribine, LEUS ⁇ TM), 5-azacitidine (azacitidine, VIDAZA®), 5-fluorouracil (5-FU, fluorouracil, ADRUCIL®), 6-mercaptopurine (6-MP, mercaptopurine, PURINETHOL®), 6- TG (6-thioguanine, thioguanine, THIO GUANINE TABLOID®), abraxane (paclitaxel protein-bound), actinomycin-D (dactinomycin, COSMEGEN®), alitretinoin (PANRETIN
- VEPESID® EFUDEX®, FLUOROPLEX®
- VEPESID® EFUDEX®, FLUOROPLEX®
- VUMON® VUMON®
- TESPA thiophosphoamide, thiotepa, TSPA, THIOPLEX®
- topotecan
- HYCAMTIN® vinblastine (vinblastine sulfate, vincaleukoblastine, VLB, ALKABAN- AQ®, VELBAN®), vinorelbine (vinorelbine tartrate, NAVELBINE®), and vorinostat (ZOLINZA®).
- the AHCM agent and/or the microenvironment modulator is administered in conjunction with a biologic.
- a biologic e.g., a binding molecule of the invention may be administered, for example, in conjunction with such known biologies.
- HERCEPTIN® trastuzumab, Genentech Inc., South San Francisco, Calif; a humanized monoclonal antibody that has anti-tumor activity in HER2 -positive breast cancer
- FASLODEX® fullvestrant, AstraZeneca Pharmaceuticals, LP, Wilmington, Del.; an estrogen-receptor antagonist used to treat breast cancer
- ARIMIDEX® anastrozole, AstraZeneca Pharmaceuticals, LP; a nonsteroidal aromatase inhibitor which blocks aromatase, an enzyme needed to make estrogen
- Aromasin® exemestane, Pfizer Inc., New York, N.Y.; an irreversible, steroidal aromatase inactivator used in the treatment of breast cancer
- FEMARA® letrozole, Novartis Pharmaceuticals, East Hanover, N.J.; a nonsteroidal aromatase inhibitor approved by the FDA
- AVASTIN® bevacizumab, Genentech Inc.; the first FDA-approved therapy designed to inhibit angiogenesis
- ZEVALIN® ibritumomab tiuxetan, Biogen personal, Cambridge, Mass.; a radiolabeled monoclonal antibody currently approved for the treatment of B-cell lymphomas.
- AVASTIN® avian avian
- ERBITUX® cetuximab, ImClone Systems Inc., New York, N.Y., and Bristol-Myers Squibb, New York, N.Y.
- EGFR epidermal growth factor receptor
- GLEEVEC® imatinib mesylate; a protein kinase inhibitor
- ERGAMISOL® levamisole hydrochloride, Janssen Pharmaceutica Products, LP, Titusville, N.J.; an immunomodulator approved by the FDA in 1990 as an adjuvant treatment in combination with 5-fluorouracil after surgical resection in patients with Dukes' Stage C colon cancer.
- exemplary biologies include TARCEVA® (erlotinib HCL, OSI Pharmaceuticals Inc., Melville, N.Y.; a small molecule designed to target the human epidermal growth factor receptor 1 (HER1) pathway).
- TARCEVA® erlotinib HCL, OSI Pharmaceuticals Inc., Melville, N.Y.
- HER1 human epidermal growth factor receptor 1
- exemplary biologies include
- VELCADE® Velcade (bortezomib, Millennium Pharmaceuticals, Cambridge Mass.; a proteasome inhibitor). Additional biologies include THALIDOMID® (thalidomide, Clegene Corporation, Warren, N.J.; an immunomodulatory agent and appears to have multiple actions, including the ability to inhibit the growth and survival of myeloma cells and anti- angiogenesis).
- Additional exemplary cancer therapeutic antibodies include, but are not limited to, 3F8, abagovomab, adecatumumab, afutuzumab, alacizumab pegol, alemtuzumab (CAMPATH®, MABCAMPATH®), altumomab pentetate (H YBRI- CEAKER®), anatumomab mafenatox, anrukinzumab (IMA-638), apolizumab, arcitumomab (CEA- SCAN®), bavituximab, bectumomab (LYMPHOSCAN®), belimumab (BENLYSTA®, LYMPHOSTAT-B®), besilesomab (SCiNTiMUN®), bevacizumab (A VASTEST®), bivatuzumab mertansine, blinatumomab, brentuximab vedotin
- ertumaxomab (REXOMUN®), etaracizumab, farletuzumab, figitumumab, fresolimumab, galiximab, gemtuzumab ozogamicin (MYLOTARG®), girentuximab, glembatumumab vedotin, ibritumomab (ibritumomab tiuxetan, ZEVALIN®), igovomab (INDIMACIS- 125®), intetumumab, inotuzumab ozogamicin, ipilimumab, iratumumab, labetuzumab (CEA- CIDE®), lexatumumab, lintuzumab, lucatumumab, lumiliximab, mapatumumab, mapatumumab, mapatumumab, mapatumumab,
- the AHCM and/or the microenvironment modulator is administered in combination with a viral cancer therapeutic agent.
- viral cancer therapeutic agents include, but not limited to, vaccinia virus (vvDD-CDSR),
- carcinoembryonic antigen-expressing measles virus recombinant vaccinia virus (TK-deletion plus GM-CSF), Seneca Valley virus-001, Newcastle virus, coxsackie virus A21, GL-ONC1, EBNA1 C-terminal LMP2 chimeric protein-expressing recombinant modified vaccinia Ankara vaccine, carcinoembryonic antigen-expressing measles virus, G207 oncolytic virus, modified vaccinia virus Ankara vaccine expressing p53, OncoVEX GM-CSF modified herpes-simplex 1 virus, fowlpox virus vaccine vector, recombinant vaccinia prostate- specific antigen vaccine, human papillomavirus 16/18 LI virus-like particle/AS04 vaccine, MVA- EBNA1/LMP2 Inj.
- TK-deletion plus GM-CSF Seneca Valley virus-001, Newcastle virus, coxsackie virus A21, GL-ONC1,
- nanopharmaceuticals include, but not limited to, ABRAXANE® (paclitaxel bound albumin nanoparticles), CRLXIO I (CPT conjugated to a linear cyclodextrin-based polymer), CRLX288 (conjugating docetaxel to the biodegradable polymer poly (lactic-co-glycolic acid)), cytarabine liposomal (liposomal Ara-C, DEPOCYTTM), daunorubicin liposomal (DAUNOXOME®), doxorubicin liposomal (DOXIL®, CAELYX®), encapsulated- daunorubicin citrate liposome (DAUNOXOME®), and PEG anti-VEGF aptamer
- the AHCM agent and/or the microenvironment modulator is administered in combination with paclitaxel or a paclitaxel formulation, e.g., TAXOL®, protein-bound paclitaxel (e.g., ABRAXANE®).
- a paclitaxel formulation e.g., TAXOL®, protein-bound paclitaxel (e.g., ABRAXANE®).
- Exemplary paclitaxel formulations include, but are not limited to, nanoparticle albumin-bound paclitaxel (ABRAXANE®, marketed by Abraxis Bioscience), docosahexaenoic acid bound-paclitaxel (DHA-paclitaxel, Taxoprexin, marketed by Protarga), polyglutamate bound-paclitaxel (PG- paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX, marketed by Cell Therapeutic), the tumor- activated prodrug (TAP), ANG105 (Angiopep-2 bound to three molecules of paclitaxel, marketed by ImmunoGen), paclitaxel-EC-1 (paclitaxel bound to the erbB2- recognizing peptide EC- 1; see Li et al, Biopolymers (2007) 87:225-230), and glucose- conjugated paclitaxel ⁇ e.g., 2'-paclitaxel methyl 2-glucopyrano
- RNAi and antisense RNA agents for treating cancer include, but not limited to, CALAA-01, siG12D LODER (Local Drug EluteR), and ALN-VSP02.
- cancer therapeutic agents include, but not limited to, cytokines (e.g., aldesleukin (IL-2, Interleukin-2, PROLEU IN®), alpha Interferon (IFN-alpha, Interferon alfa, INTRON® A (Interferon alfa-2b), ROFERON-A® (Interferon alfa-2a)), Epoetin alfa (PROCRIT®), filgrastim (G-CSF, Granulocyte - Colony Stimulating Factor,
- cytokines e.g., aldesleukin (IL-2, Interleukin-2, PROLEU IN®
- IFN-alpha Interferon alfa
- INTRON® A Interferon alfa-2b
- ROFERON-A® Interferon alfa-2a
- Epoetin alfa PROCRIT®
- filgrastim G-CSF, Granulocyte - Colony Stimulating Factor
- GM-CSF Granulocyte Macrophage Colony Stimulating Factor, sargramostim, LEUKINETM
- IL-1 1 Interleukin- 1 1, oprelvekin, NEUMEGA®
- Interferon alfa-2b PEG conjugate
- PEG interferon PEG-INTRONTM
- pegfilgrastim PEG interferon, PEG-INTRONTM
- hormone therapy agents e.g., aminoglutethimide (CYTADREN®), anastrozole (ARIMIDEX®), bicalutamide (CASODEX®), exemestane (AROMASIN®), fluoxymesterone (HALOTESTIN®), flutamide (EULEXIN®), fulvestrant (FASLODEX®), goserelin (ZOLADEX®), letrozole (FEMARA®), leuprolide (ELIGARDTM, LUPRON®, LUPRON DEPOT®, VIADURTM), megestrol (megestrol acetate, MEGACE®), nilutamide (ANANDRON®, NILANDRON®), octreotide (octreotide acetate, SANDOSTATIN®, SANDOSTATIN LAR®), raloxifene (EVISTA®), romiplostim (NPLATE®), tamoxifen (NOVALDEX
- GLEEVECTM lapatinib (TY ERB®), sorafenib (NEXAVAR®), and SU11248 (sunitinib, SUTENT®)), immunomodulatory and antiangiogenic agents (e.g., CC-5013 (lenalidomide, REVLIMID®), and thalidomide (THALOMID®)), glucocorticosteroids (e.g., cortisone (hydrocortisone, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, ALA- CORT®, HYDROCORT ACETATE®, hydrocortone phosphate LANACORT®, SOLU- CORTEF®), decadron (dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, DEXASONE®, DIODEX®, HEXADROL®, MAXIDEX®), methylprednisolone (6-methylprednisolone
- the AHCM agent and/or the microenvironment modulator is used in combination with a tyrosine kinase inhibitor (e.g., a receptor tyrosine kinase (RTK) inhibitor).
- a tyrosine kinase inhibitor e.g., a receptor tyrosine kinase (RTK) inhibitor.
- Exemplary tyrosine kinase inhibitor include, but are not limited to, an epidermal growth factor (EGF) pathway inhibitor (e.g., an epidermal growth factor receptor (EGFR) inhibitor), a vascular endothelial growth factor (VEGF) pathway inhibitor (e.g., an antibody against VEGF, a VEGF trap, a vascular endothelial growth factor receptor (VEGFR) inhibitor (e.g., a VEGFR- 1 inhibitor, a VEGFR-2 inhibitor, a VEGFR-3 inhibitor)), a platelet derived growth factor (PDGF) pathway inhibitor (e.g., a platelet derived growth factor receptor (PDGFR) inhibitor (e.g., a PDGFR- ⁇ inhibitor)), a RAF- 1 inhibitor, a KIT inhibitor and a RET inhibitor.
- the anti-cancer agent used in combination with the AHCM agent is selected from the group consisting of: axitinib
- VECTIBIX® ranibizumab
- TASIGNA® nilotinib
- NEXAVAR® alemtuzumab
- CAMPATH® gemtuzumab ozogamicin
- MYLOTARG® ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOKTM), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154, CEP-1 1981, tivozanib (AV-951), OSI-930, MM- 121, XL-184, XL-647, XL228, AEE788, AG-490, AST-6, BMS-599626, CUDC-101, PD153035, pelitinib (EKB- 569), vande
- Selected tyrosine kinase inhibitors are chosen from sunitinib, erlotinib, gefitinib, or sorafenib. In one embodiment, the tyrosine kinase inhibitor is sunitinib.
- the AHCM and/or the microenvironment modulator is administered in combination with one of more of: an anti-angiogenic agent, or a vascular targeting agent or a vascular disrupting agent.
- anti-angiogenic agents include, but are not limited to, VEGF inhibitors (e.g., anti-VEGF antibodies (e.g., bevacizumab); VEGF receptor inhibitors (e.g., itraconazole); inhibitors of cell proliferatin and/or migration of endothelial cells (e.g., carboxyamidotriazole, TNP-470); inhibitors of angiogenesis stimulators (e.g., suramin), among others.
- VEGF inhibitors e.g., anti-VEGF antibodies (e.g., bevacizumab); VEGF receptor inhibitors (e.g., itraconazole); inhibitors of cell proliferatin and/or migration of endothelial cells (e.g., carboxyamidotriazole, T
- VTA vascular-targeting agent
- VDA vascular disrupting agent
- VTAs can be small-molecule.
- Exemplary small-molecule VTAs include, but are not limited to, microtubule destabilizing drugs (e.g., combretastatin A-4 disodium phosphate (CA4P), ZD6126, AVE8062, Oxi 4503); and vadimezan (ASA404).
- microtubule destabilizing drugs e.g., combretastatin A-4 disodium phosphate (CA4P), ZD6126, AVE8062, Oxi 4503
- ASA404 vadimezan
- radioisotopes include: 90 Y, 125 I, 131 I, 123 I, m In, 105 Rh, 153 Sm, 67 Cu, 67 Ga, 166 Ho, 177 Lu, 186 Re and 188 Re.
- the radionuclides act by producing ionizing radiation which causes multiple strand breaks in nuclear DNA, leading to cell death.
- the isotopes used to produce therapeutic conjugates typically produce high energy -or ⁇ -particles which have a short path length. Such radionuclides kill cells to which they are in close proximity, for example neoplastic cells to which the conjugate has attached or has entered. They have little or no effect on non-localized cells. Radionuclides are essentially non- immunogenic.
- binding molecules can be conjugated to different radiolabels for diagnostic and therapeutic purposes.
- the aforementioned U.S. Pat. Nos. 6,682, 134, 6,399,061, and 5,843,439 disclose radiolabeled therapeutic conjugates for diagnostic "imaging" of tumors before administration of therapeutic antibody.
- “In2B8" conjugate comprises a murine monoclonal antibody, 2B8, specific to human CD20 antigen, that is attached to 11 'in via a bifunctional chelator, i.e., MX-DTPA (diethylenetriaminepentaacetic acid), which comprises a 1 : 1 mixture of l-isothiocyanatobenzyl-3-methyl-DTPA and l-methyl-3-isothiocyanatobenzyl- DTPA.
- 11 'in is particularly preferred as a diagnostic radionuclide because between about 1 to about 10 mCi can be safely administered without detectable toxicity; and the imaging data is generally predictive of subsequent 90 Y-labeled antibody distribution.
- the cancer therapy includes an immune therapy used in combination with the AHCM, other cancer therapies, and/or the microenvironment modulator, described herein.
- an immune therapy used in combination with the AHCM, other cancer therapies, and/or the microenvironment modulator, described herein.
- hypoxia and/or limited perfusion are believed to cause immunosuppression and/or limit the efficacy of certain immune therapies.
- AHCM alone or in combination with therapies described herein can be used to improve the efficacy of said immune therapies.
- immune therapies include, but are not limited to, CTLA-4 blockade ⁇ e.g., an anti-CTLA-4 antibody ⁇ e.g., ipilimumab)); immune-based therapies (including, e.g., immune or dendritic cell- based vaccines and antagonists of immune inhibitory signals or checkpoints); cancer vaccines, e.g., Sipuleucel-T (APC8015, trade name Provenge, manufactured by Dendreon Corporation) is a therapeutic cancer vaccine for prostate cancer (CaP)); and adoptive T-cell-based therapies.
- Exemplary immune-based therapies include, but are not limited to, e.g., immune or dendritic cell- based vaccines (Seton-Rogers, S.
- the cancer therapy includes PDT used in combination with the AHCM, other cancer therapies, and/or the microenvironment modulator, described herein.
- PDT includes administration of a photosensitizing agent (e.g., a porhyrin, a porpyrin precursor, a chorlin, or a phthalocyanine) followed by irradiation at a wavelength corresponding to an absorbance band of the sensitizer.
- a photosensitizing agent e.g., a porhyrin, a porpyrin precursor, a chorlin, or a phthalocyanine
- cell death e.g., tumor cell death
- damage to the microvasculature e.g., damage to the microvasculature
- induction of a local inflammatory reaction e.g., Agostinis, P. et al. (2011) CA Cancer J. Clin. 61 :250- 281.
- the cancer therapy includes an inhibitor of a cancer stem cell (also referred to herein as a "cancer initiating cell”), used in combination with the AHCM, other cancer therapies and/or the microenvironment modulator, described herein.
- a cancer stem cell also referred to herein as a "cancer initiating cell”
- hypoxia and cancer drugs including anti- angiogenic drugs
- radiation therapy are believed to increase the number of cancer stem cells.
- AHCM AHCM
- an inhibitor of a cancer stem cell can be used to reduce the production of these stem cells.
- exemplary inhibitors of cancer stem cells include, but are not limited to, hedgehog (e.g., SMO) antagonists; and Wnt pathway antagonists (e.g., antibody, OMP- 18R5).
- the AHCM agent, the microenvironment modulator and/or the additional anti-cancer agent are administered concurrently (e.g., administration of the two agents at the same time or day, or within the same treatment regimen) and/or sequentially (e.g., administration of one agent over a period of time followed by
- the AHCM and/or the microenvironment modulator is administered prior to the anti-cancer agent. In other embodiments, the AHCM and/or the microenvironment modulator is administered prior to the anti-cancer agent, and followed by concurrent administration of the AHCM, the microenvironment modulator and/or the anticancer agent.
- the AHCM agent, the microenvironment modulator and/or and the additional anti-cancer agent are administered concurrently.
- the AHCM agent, the microenvironment modulator and/or and the additional anti-cancer agent are administered at the same time, on the same day, or within the same treatment regimen.
- microenvironment modulator is administered before the additional anti-cancer agent on the same day or within the same treatment regimen.
- the AHCM agent and/or the microenvironment modulator is concurrently administered with additional anti-cancer agent for a period of time, after which point treatment with the additional anti-cancer agent is stopped and treatment with the AHCM agent continues.
- the AHCM agent and/or the microenvironment modulator is concurrently with the additional anti-cancer agent for a period of time, after which point treatment with the AHCM agent and/or the microenvironment modulator is stopped and treatment with the additional anti-cancer agent continues.
- the AHCM agent, the microenvironment modulator and/or the additional anti-cancer agent are administered sequentially.
- the AHCM agent is administered after the treatment regimen of the additional anti-cancer agent and/or microenvironment modulator has ceased.
- the additional anti-cancer agent is administered after the treatment regimen of the AHCM agent and/or microenvironment modulator has ceased.
- the AHCM agent, microenvironment modulator and/or the anti-cancer agent can be administered in a pulse administration.
- they can be administered as a pulse-chase administration, e.g., where an AHCM agent is administered for a brief period of time (pulse), followed by administration of an anti-cancer agent for a longer period of time (e.g., chase), or vice versa.
- pulse-chase administration e.g., where an AHCM agent is administered for a brief period of time (pulse), followed by administration of an anti-cancer agent for a longer period of time (e.g., chase), or vice versa.
- AHCM agents can be used to improve diagnosis, treatment, prevention and/or prognosis of cancers in mammals, preferably humans.
- diagnostic assays can be performed in vivo or in vitro, such as, for example, on blood samples, biopsy tissue or autopsy tissue.
- the invention provides a diagnostic method useful during diagnosis of a cancer, which involves measuring the expression level of target protein or transcript in tissue or other cells or body fluid from an individual and comparing the measured expression level with a standard target expression levels in normal tissue or body fluid, whereby an increase in the expression level compared to the standard is indicative of a disorder.
- One embodiment provides a method of detecting the presence of abnormal hyperproliferative cells, e.g., precancerous or cancerous cells, in a fluid or tissue sample, comprising assaying for the expression of the target in tissue or body fluid samples of an individual and comparing the presence or level of target expression in the sample with the presence or level of target expression in a panel of standard tissue or body fluid samples, where detection of target expression or an increase in target expression over the standards is indicative of aberrant hyperproliferative cell growth.
- abnormal hyperproliferative cells e.g., precancerous or cancerous cells
- One aspect of the invention is a method for the in vivo detection or diagnosis of a cancer in a subject, preferably a mammal and most preferably a human.
- diagnosis comprises: a) administering (for example, parenterally,
- radioactivity injected will normally range from about 5 to 20 millicuries of, e.g., 99 Tc.
- the labeled binding molecule e.g., antibody or antibody fragment
- In vivo tumor imaging is described in S. W. Burchiel et al., "Immunopharmacokinetics of Radiolabeled Antibodies and Their Fragments.” (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., Masson Publishing Inc. (1982).
- the time interval following the administration for permitting the labeled molecule to preferentially concentrate at sites in the subject and for unbound labeled molecule to be cleared to background level is 6 to 48 hours or 6 to 24 hours or 6 to 12 hours. In another embodiment the time interval following administration is 5 to 20 days or 7 to 10 days.
- Presence of the labeled molecule can be detected in the patient using methods known in the art for in vivo scanning. These methods depend upon the type of label used. Skilled artisans will be able to determine the appropriate method for detecting a particular label. Methods and devices that may be used in the diagnostic methods of the invention include, but are not limited to, computed tomography (CT), whole body scan such as position emission tomography (PET), magnetic resonance imaging (MRI), and sonography, X- radiography, nuclear magnetic resonance imaging (NMR), CAT-scans or electron spin resonance imaging (ESR).
- CT computed tomography
- PET position emission tomography
- MRI magnetic resonance imaging
- sonography sonography
- X- radiography nuclear magnetic resonance imaging
- NMR nuclear magnetic resonance imaging
- CAT-scans or electron spin resonance imaging (ESR).
- compositions described herein can be incorporated into a variety of formulations for administration. More particularly, the compositions can be formulated into pharmaceutical compositions by combination with appropriate, pharmaceutically acceptable carriers or diluents, and can be formulated into preparations in semi-solid, liquid or gaseous forms; such as capsules, powders, granules, gels, slurries, ointments, solutions, suppositories, injections, inhalants and aerosols. As such, administration of the compositions can be achieved in various ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, intratracheal administration. Moreover, the compositions can be administered in a local rather than systemic manner, in a depot or sustained release formulation.
- compositions can be formulated with common excipients, diluents or carriers, and compressed into tablets, or formulated as elixirs or solutions for convenient oral administration, or administered by the intramuscular or intravenous routes.
- the compositions can be administered transdermally, and can be formulated as sustained release dosage forms and the like.
- Compositions can be administered alone, in combination with each other, or they can be used in combination with other known compounds (discussed herein).
- compositions described herein can be manufactured in a manner that is known to those of skill in the art, e.g., by mixing, dissolving, granulating, dragee-making, levigating, emulsifying,
- compositions can be formulated by combining with pharmaceutically acceptable carriers that are known in the art.
- Such carriers enable the compounds to be formulated as pills, capsules, emulsions, lipophilic and hydrophilic suspensions, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated.
- Pharmaceutical preparations for oral use can be obtained by mixing the compositions with an excipient and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
- Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium
- carboxymethylcellulose and/or polyvinylpyrrolidone (PVP).
- PVP polyvinylpyrrolidone
- compositions for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g.,
- the dosage unit can be determined by providing a valve to deliver a metered amount.
- Capsules and cartridges of, e.g., gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
- compositions can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
- Formulations for injection can be presented in unit dosage form, e.g., in ampules or in multidose containers, with an added preservative.
- the compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulator agents such as suspending, stabilizing and/or dispersing agents.
- compositions can also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter, carbowaxes, polyethylene glycols or other glycerides, all of which melt at body temperature, yet are solidified at room temperature.
- rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter, carbowaxes, polyethylene glycols or other glycerides, all of which melt at body temperature, yet are solidified at room temperature.
- compositions can also be formulated as a depot preparation.
- long acting formulations can be administered by implantation (for example
- the compounds can be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
- Lipid particles e.g., liposomes
- emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Long-circulating, e.g., stealth, liposomes can be employed. Such liposomes are generally described in U.S. Pat. No. 5,013,556.
- the compounds of the present invention can also be administered by controlled release means and/or delivery devices such as those described in U.S. Pat. Nos. 3,845,770; 3,916,899;
- compositions suitable for use in the present invention include compositions wherein the active ingredients are contained 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
- a suitable daily dose of an AHCM agent and/or a cancer therapeutic can be that amount of the compound which is the lowest dose effective to produce a therapeutic effect.
- Such an effective dose can generally depend upon the factors described above.
- the subject receiving this treatment is any animal in need, including primates, in particular humans, equines, cattle, swine, sheep, poultry, dogs, cats, mice and rats.
- the compounds can be administered daily, every other day, three times a week, twice a week, weekly, or bi-weekly.
- the dosing schedule can include a "drug holiday," i.e., the drug can be administered for two weeks on, one week off, or three weeks on, one week off, or four weeks on, one week off, etc., or continuously, without a drug holiday.
- the compounds can be administered orally, intravenously, intraperitoneally, topically, transdermally, intramuscularly, subcutaneously, intranasally, sublingually, or by any other route.
- the AHCM agents are administered in combination with other treatments (such as additional chemotherapeutics, radiation or surgery) the doses of each agent or therapy can be lower than the corresponding dose for single-agent therapy.
- additional chemotherapeutics, radiation or surgery the doses of each agent or therapy can be lower than the corresponding dose for single-agent therapy.
- the determination of the mode of administration and the correct dosage is well within the knowledge of the skilled clinician.
- the AHCM (alone or in combination with the microenvironment modulator and/or cancer therapy) is formulated for oral, subcutaneous, intravenous or intraperitoneal administration.
- the AHCM (alone or in combination with the microenvironment modulator and/or cancer therapy) is formulated for oral administration (e.g., an oral tablet or pill).
- substantially continuous administration of an AHCM causes a greater reduction in collagen content and/or tumor size than single or pulsatile administration of the AHCM.
- it may be desirable to formulate and/or administered the AHCM (alone or in combination with the microenvironment modulator and/or cancer therapy) substantially continuously.
- the AHCM (alone or in combination) is administered substantially continuously over a pre-determined period of, or at least 15, 30, 45 minutes; a period of, or at least, 1, 5, 10, 24 hours; a period of, or at least, 2, 5, 10, 14 days; a period of, or at least, 3, 4, 5, 6, 7, 8 weeks; a period of, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 months; a period of, or at least, 1, 2, 3, 4, 5 years, or longer.
- the delivery method can be optimized such that an AHCM dose as described herein (alone or in combination) is administered and/or maintained in the subject for a pre-determined period (e.g., a period as described herein).
- the AHCM (alone or in combination with the microenvironment modulator and/or cancer therapy) is in a controlled- or extended release formulation, dosage form, or device.
- exemplary formulations and devices for controlled or extended release are known in the art.
- hydroxypropylmethyl cellulose gels, osmotic systems, liposomes and combination thereof can be used to provide the desired release kinetics.
- the AHCM is administered via an implantable infusion device, e.g., a pump (e.g., a subcutaneous pump), an implant or a depot.
- Implantable infusion devices typically include a housing containing a liquid reservoir which can be filled transcutaneously by a hypodermic needle penetrating a fill port septum. The medication reservoir is generally coupled via an internal flow path to a device outlet port for delivering the liquid through a catheter to a patient body site.
- Typical infusion devices also include a controller and a fluid transfer mechanism, such as a pump or a valve, for moving the liquid from the reservoir through the internal flow path to the device's outlet port.
- AHCM agents described herein the anti-cancer agents (e.g., low molecular weight, mid-molecular weight anti-cancer agents described herein), or both, can be packaged in nanoparticles.
- nanoparticles are from 10, 15, 20, 25, 30, 35, 45, 50, 75, 100, 150 or 200 nm or 200- 1,000, e.g., 10, 15, 20, 25, 30, 35, 45, 50, 75, 100, 150, or 200, or 20 or 30 or 50-400 nm in diameter. Smaller particles tend to be cleared more rapidly form the system. Drugs can be intrapped within or coupled, e.g., covalent coupled, or otherwise adhered, to nanoparticles.
- Lipid- or oil-based nanoparticles such as liposomes and solid lipid nanoparticles and can be used to can be used to deliver agents described herein. DOXIL® is an example of a liposomic nanoparticle.
- Solid lipid nanoparticles for the delivery on anticancer agents are descripbed in Serpe et al. (2004) Eur. J. Pharm. Bioparm. 58:673-680 and Lu et al. (20060 Eur. J. Pharm. Sci. 28: 86-95.
- Polymer-based nanoparticles e.g., PLGA- based nanoparticles can be used to deliver agents described herein. These tend to rely on biodegradable backbone with the theraeutic agent intercalated (with or without covalent linkage to the polymer) in a matrix of polymer.
- PLGA is a widely used in polymeric nanoparticles, see Hu et al. (2009) J. Control.
- PEGylated PLGA-based nanoparticles can also be used to deliver anti-cancer agents, see, e.g., Danhhier et al., (2009) J. Control. Release 133 : 1 1- 17, Gryparis et al (2007) Eur. J. Pharm. Biopharm. 67: 1-8.
- Metal-based, e.g., gold-based nanoparticles can also be used to dleiver anti-cancer agents.
- Protien-based, e.g., albumin-based nanoparticles can be used to deliver agents described herein.
- an agent can be bound to nanoparticles of human albumin.
- An exemplary anti-cancer agent/protein nanoparticle is Abraxane®, in which paclitaxel is pund to nanparticles of albumin.
- Nanoparticles can employ active targeting, passive targeting or both. Active targeting can rely on inclusion of a ligand tht binds with a target at or near a preselected site, e.g., a solid tumor. Passive targeting nanoparticles can diffuse and accumalte at sites of interest, e.g., sites characterized by excessivley leaky micorvasculatiure, e.g., as seen in tumors and sites of inflammation.
- a method of treating a subject having a disorder, e.g., improving the delivery or efficacy of a therapy, in a subject comprising:
- a subject having a disorder e.g., improving the delivery or efficacy of the therapy, in a subject.
- a method of treating or preventing a cancer, in a subject comprising:
- AHCM collagen modifying agent
- AHCM and/or microenvironment modulator is administered in a dosage sufficient to treat or prevent the cancer.
- microenvironment modulator is a microenvironment modulator.
- said parameter is chosen from one or more of: a) objective response rate (ORR); b) progression free survival (PFS); c) overall survival (OS); or d) reduction in toxicity.
- a) drug concentration e.g., at a disorder or disease site, e.g., a solid tumor
- blood perfusion e.g., at a disorder or disease site, e.g., a solid tumor
- oxygenation e.g., at a disorder or disease site, e.g., a solid tumor
- interstitial fluid pressure e.g., at a disorder or disease site, e.g., a solid tumor
- extracellular matrix content or composition e.g., level of collagen and/or hyaluronic acid. 6.
- the subject has not been administered a dose of the AHCM within 5, 10, 30, 60 or 100 days of the diagnosis of the disorder (e.g., cancer) or the initiation of the AHCM dosing; c) the subject is not hypertensive, or has been hypertensive, prior to administration of the AHCM;
- the AHCM and/or microenvironment modulator is administered at least one, two, three, or five days; or one, two, three, four, five or more weeks, prior to the therapy, e.g., the cancer therapy;
- the AHCM and/or microenvironment modulator is administered at least one, two, three, or five days; or one, two, three, four, five or more weeks, prior to the therapy, e.g., the cancer therapy, and concurrently with the therapy, e.g., the cancer therapy,
- the AHCM and/or microenvironment modulator is administered continuously over a period of at least 1, 5, 10, or 24 hours; at least 2, 5, 10, or 14 days; at least 2, 3, 4, 5 or 6 weeks; at least 2, 3, 4, 5 or 6 months; or at least 1, 2, 3, 4 or 5 years, or
- the AHCM and/or microenvironment modulator is administered after cessation of the therapy, e.g., the cancer therapy, e.g., at least days, weeks, months or years after cessation of the therapy, e.g., the cancer therapy.
- an angiotensin II receptor blocker (ATi blocker),
- RAAS antagonist an antagonist of renin angiotensin aldosterone system
- an angiotensin converting enzyme (ACE) inhibitor (iii) an angiotensin converting enzyme (ACE) inhibitor
- TSP-1 thrombospondin 1
- TGF- ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- AHCM is an ATi inhibitor chosen from one or more of: losartan (COZAAR®), candesartan (ATACAND®), eprosartan mesylate (TEVETEN®), EXP 3174, irbesartan (AVAPRO®), L158,809, olmesartan (BENICAR®), saralasin, telmisartin (MICARDIS®), valsartan (DIOVAN®), or a derivative thereof.
- AHCM is a RAAS antagonist chosen from one or more of: aliskiren (TEKTURNA®, RASILEZ®), remikiren (Ro 42- 5892), enalkiren (A-64662), SPP635, or a derivative thereof.
- AHCM is an ACE inhibitor chosen from one or more of: benazepril (LOTENSIN®), captopril (CAPOTEN®), enalapril (VASOTEC®), fosinopril (MONOPRIL®), lisinopril (PRINIVIL®, ZESTRIL®), moexipril (UNIVASC®), perindopril (ACEON®), quinapril (ACCUPRIL®), ramipril (ALTACE®), trandolapril (MAVIK®), or a derivative thereof.
- benazepril LOTENSIN®
- captopril CAPOTEN®
- VASOTEC® enalapril
- VASOTEC® fosinopril
- PRINIVIL® lisinopril
- TRIVASC® moexipril
- perindopril ACEON®
- quinapril ACCUPRIL®
- ramipril trando
- AHCM is a TSP-1 inhibitor chosen from one or more of: ABT-510, CVX-045, LSKL, or a derivative thereof.
- TGF- ⁇ inhibitor is chosen from one or more of: an anti- TGF- ⁇ antibody, or a TGF- ⁇ 1 peptide inhibitor.
- CTGF inhibitor is chosen from one or more of: DN-9693, FG-3019, or a derivative thereof.
- the microenvironment modulator is chosen from one or more of an anti-angiogenic therapy; an inhibitor of vascular endothelial growth factor (VEGF) pathway; an agent that decreases the level or production of hyaluronic acid; an inhibitor of the hedgehog pathway; a disulfide-based cyclic RGD peptide peptide (iRGD) or an analogue thereof; a taxane therapy; an agent that decreases the level or production of collagen or procollagen; an anti-fibrotic agent; or a profibrotic pathway inhibitor.
- VEGF vascular endothelial growth factor
- iRGD disulfide-based cyclic RGD peptide peptide
- microenvironment modulator is administered in an amount sufficient to enhance the distribution or efficacy of the cancer therapy. 17.
- microenvironment modulator is administered at a dose that causes one or more of: decreases the level or production of collagen, decreases tumor fibrosis, reduces interstitial fluid pressure, increases interstitial tumor transport, improves tumor perfusion, increases tumor oxygenation; decreases tumor hypoxia; decreases tumor acidosis; enables immune cell infiltration; decreases immunosuppression; increases antitumor immunity; decreases cancer stem cells (also referred to herein as tumor-initiating-cells); or enhances penetration or diffusion, of the cancer therapy in a tumor or tumor vasculature, in the subject.
- microenvironment modulator or the cancer therapeutic, each independently, is provided as an entity having the following size ranges (in nm): a hydrodynamic diameter of less than or equal to 1, or between 0.1 and 1.0 nm; a hydrodynamic diameter of between 5 and 20, or 5 and 15 nm; or a hydrodynamic diameter of 1, 5, 10, 15, 20, 25, 30, 35, 45, 50, 75, 100, 150, 200 nm, but less than 300 nm.
- cell-based immunotherapy comprises expanding immune cells, e.g., ex vivo, and injecting the expanded immune cells into the subject.
- cancer therapy is chosen from one or more of anti-cancer agents, photodynamic therapy, an immunotherapy (e.g., an immune-cell therapy or adoptive immunotherapy), surgery and/or radiation.
- an immunotherapy e.g., an immune-cell therapy or adoptive immunotherapy
- a cancer therapeutic chosen from a viral cancer therapeutic agent, a lipid nanoparticle of an anti-cancer therapeutic agent, a polymeric nanoparticle of an anti-cancer therapeutic agent, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a chemotherapeutic agent;
- an immunotherapy e.g., an immune-cell therapy or adoptive immunotherapy
- lipid nanoparticle is chosen from pegylated liposomal doxorubicin (DOXIL ® ) or liposomal paclitaxel (e.g., Abraxane®).
- chemotherapeutic agent is chosen from gemcitabine, cisplatin, epirubicin, 5-fluorouracil, paclitaxel, oxaliplatin, or leucovorin.
- cancer therapy is a tyrosine kinase inhibitor chosen from sunitinib, erlotinib, gefitinib, sorafenib, icotinib, lapatinib, neratinib, vandetanib, BIBW 2992 or XL-647, or an anti-EGFR antibody chosen from cetuximab, panitumumab, zalutumumab, nimotuzumab necitumumab or matuzumab.
- tyrosine kinase inhibitor chosen from sunitinib, erlotinib, gefitinib, sorafenib, icotinib, lapatinib, neratinib, vandetanib, BIBW 2992 or XL-647
- an anti-EGFR antibody chosen from cetuximab, panitumumab, zalutumumab, nimot
- chemotherapeutic agent is a cytotoxic or a cytostatic agent.
- chemotherapeutic agent is chosen from an antimicrotubule agent, a topoisomerase inhibitor, a taxane, an antimetabolite, a mitotic inhibitor, an alkylating agent, or an intercalating agent.
- cancer therapy is chosen from one of more of: an anti- angiogenic agent, or a vascular targeting agent or a vascular disrupting agent.
- microenvironment modulator or the cancer therapy is administered to the subject by a systemic administration chosen from oral, parenteral, subcutaneous, intravenous, rectal, intramuscular, intraperitoneal, intranasal, transdermal, or by inhalation or intracavitary installation.
- TGFpi transforming growth factor beta 1
- CGF connective tissue growth factor
- TSP-1 thrombospondin- 1
- angiotensin receptor the level or expression of an angiotensin receptor
- a biomarker chosen from collagen I, collagen III, collagen IV, TGFpi, CTGF, or
- tumor aggressivity tumor aggressivity, vascularization of primary tumor, or metastatic spread.
- a pharmaceutical composition comprising a nanoparticle comprising an AHCM.
- a pharmaceutical composition comprising a nanoparticle comprising an AHCM, a microenvironment modulator, and/or a cancer therapeutic agent.
- the cancer therapeutic is chosen from a viral cancer therapeutic agent, a lipid nanoparticle of an anti-cancer agent, a polymeric nanoparticle of an anti-cancer agent, an antibody against a cancer target, a dsRNA agent, an antisense RNA agent, or a chemotherapeutic agent.
- a dosage form of an AHCM wherein the AHCM is formulated in a dosage form that is less than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.16, 0.2, 0.3,
- a dosage form of an AHCM wherein the AHCM is formulated in a dosage form that is greater than 1.1, 1.5, 1.7, 2, 3, 4, 5, 10-fold or higher, that of the standard of care dosage form for anti-hypertensive or anti-heart failure use of the AHCM.
- a method optimizing access to a cancer, or optimizing delivery to a cancer of an agent comprising:
- AHCM collagen modifying agent
- administering comprises one or more of the following:
- the diagnostic or imaging agent has a hydrodynamic diameter of greater than
- the agent is a radiologic agent, an NMR agent, a contrast agent; or c) the subject is treated with a dosing of AHCM administration, whic is initiated prior to administration of the agent for at least two, three, or five days, or one, two, three, four, five or more weeks prior to administration of the agent.
- the detected change includes one or more of: an increase or decrease of activated TGF beta, TGF beta 1 level, connective tissue growth factor (CTGF) level, or collagen (e.g., collagen 1) level.
- CTGF connective tissue growth factor
- collagen e.g., collagen 1
- the candidate agent is chosen from one or more of: an antagonist of renin angiotensin aldosterone system (“RAAS antagonist”), an angiotensin converting enzyme (ACE) inhibitor, an angiotensin II receptor blocker (ATi blocker), a thrombospondin 1 (TSP-1) inhibitor, a transforming growth factor beta 1 (TGF- ⁇ ⁇ ) inhibitor, or a connective tissue growth factor (CTGF) inhibitor.
- RAAS antagonist an antagonist of renin angiotensin aldosterone system
- ACE angiotensin converting enzyme
- ATi blocker an angiotensin II receptor blocker
- TSP-1 thrombospondin 1
- TGF- ⁇ ⁇ transforming growth factor beta 1
- CTGF connective tissue growth factor
- AHCM anti-hypertensive and/or a collagen modifying
- a therapeutic kit comprising an anti-hypertensive and/or a collagen modifying (AHCM), alone or in combination with a microenvironment modulator, and/or a therapy, e.g., a cancer therapy, and instructions for use for the treatment of cancer.
- a diagnostic kit comprising an anti-hypertensive and/or a collagen modifying (AHCM), alone or in combination with an imaging agent, and instructions for use for the diagnosis of cancer.
- a method of selecting a subject for receiving an anti-hypertensive and/or a collagen modifying agent comprising:
- AHCM AHCM is administered in a dosage sufficient to improve the delivery or efficacy of the cancer therapy.
- AHCM agent for example, losartan - a clinically approved angiotensin II receptor antagonist generally for treatment of hypertension - can enhance the penetration and efficacy of nanomedicine, e.g., via an anti-fibrotic effect.
- nanotherapeutics have offered new hope for cancer treatment, 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).
- the dense collagen network in tumors can generally reduce the penetration and efficacy of nanotherapeutics. This is partly because their penetration is hindered specially in fibrotic tumors where the small interfibrillar spacing in the interstitium retards 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 Set USA 98:4628-4633;
- Matrix modifiers like bacterial collagenase, relaxin, and matrix metalloproteinase -1 and -8 have been used to modify the collagen or proteoglycan network in tumors and have improved the efficacy of intratumorally (i.t.) injected oncolytic viruses (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;
- relaxin can improve transport through the tumor matrix, but may not facilitate the delivery of low molecular weight agents (US 6,719,977). However, these agents may produce normal tissue toxicity (e.g., bacterial collagenase) or increase the risk of tumor progression (e.g., relaxin, matrix metalloproteinases).
- Losartan (Johnston CI (1995) Lancet 346: 1403-1407) - approved to control hypertension in patients - does not have many of these safety risks. Furthermore, in addition to its antihypertensive properties, losartan is also an antifibrotic agent that has been shown to reduce the incidence of cardiac and renal fibrosis (Habashi JP, et al. (2006) Science 312: 1 17- 121; and. Cohn RD, et al. (2007) Nat Med 13:204-210).
- TGF- ⁇ active transforming growth factor- ⁇
- AGTR1 angiotensin II type I receptor
- TGF- ⁇ activators like thrombospondin- 1 (TSP- 1)
- Habashi JP et al. (2006) Science 312: 117-121; Cohn RD, et al. (2007) Nat Med 13:204-210; Lavoie P, et al. (2005) JHypertens 23 : 1895- 1903; Chamberlain JS (2007) Nat Med 13 : 125-126; and Dietz HC (2010) J Clin Invest 120:403-407.
- an AHCM agent e.g., losartan inhibited collagen I production by carcinoma associated fibroblasts (CAFs) isolated from breast cancer biopsies. Additionally, an AHCM agent, e.g., losartan, led to a dose-dependent reduction in stromal collagen in desmoplastic models of human breast, pancreatic and skin tumors in mice.
- CAFs carcinoma associated fibroblasts
- an AHCM agent e.g., losartan improved the distribution and therapeutic efficacy of intratumorally injected oncolytic herpes simplex viruses (HSV). Further, an AHCM agent, e.g., losartan also enhanced the efficacy of intravenously injected pegylated liposomal doxorubicin (DOXIL ® ). Accordingly, administration of an AHCM agent, e.g., losartan, in combination with a cancer therapeutic (e.g., a cancer nanotherapeutic) can enhance the efficacy of nanotherapeutics in patients with desmoplastic tumors.
- a cancer therapeutic e.g., a cancer nanotherapeutic
- an AHCM agent e.g., losartan reduces collagen I levels in four tumor models - a spontaneous mouse mammary carcinoma (FVB MMTV PyVT), an orthotopic pancreatic adenocarcinoma (L3.6pl), and subcutaneously implanted fibrosarcoma (HSTS26T) and melanoma (Mu89).
- an AHCM agent e.g., Losartan
- an AHCM agent e.g., losartan
- AHCM agent e.g., losartan
- the inventors assessed how an AHCM agent, e.g., losartan, can affect the distribution and efficacy of oncolytic HSV administered i.t. - a widely used method of administration in patients 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) - and the efficacy of i.v.-administered DOXIL ® .
- an AHCM agent e.g., Losartan
- DOXIL ® i.t.-administered
- an AHCM agent e.g., losartan
- an AHCM agent can enhance nanoparticle penetration in the interstitial space by improving interstitial transport.
- an AHCM agent e.g., losartan
- an FDA approved antihypertensive drug can be used to improve the efficacy of various nanotherapeutics in multiple tumor types.
- Example 1 Losartan inhibits collagen I synthesis by carcinoma associated fibroblasts
- TSP-1 is a key regulator of TGF- ⁇ activation and losartan has been reported to reduce TSP-1 expression and TGF- ⁇ activation in mouse models of Marfan' s syndrome and muscular dystrophy (Dietz HC (2010) J Clin Invest 120:403 ⁇ -07).
- losartan did not affect total TGF- ⁇ levels but significantly reduced TSP-1, active TGF- ⁇ , and collagen I levels (Fig. 6).
- Losartan also decreased the TSP-1 immunostaining in HSTS26T (73% p ⁇ 0.04) and Mu89 (24% p ⁇ 0.03) (Figs. 7A-7B).
- losartan improved nanoparticle accumulation and penetration in the tumor center (Fig. 8A; HSTS26T pO.001, Mu89 p ⁇ 0.001). Conversely, there was little or no nanoparticle accumulation in the center of control tumors. Most of the injected nanoparticles in control tumors were found in the tumor margin and around the needle insertion point (Fig. 8A). The inventors also determined the effects of losartan on the intratumoral distribution of oncolytic HSV. In both HSTS26T and Mu89, losartan significantly increased the intratumoral spread of HSV injected intratumorally (Fig. 8B).
- the inventors then determined if losartan could improve the efficacy of i.t. injected oncolytic HSV and i.v. injected DOXIL®.
- the effect of losartan combined with the i.t. injection of HSV was determined in HSTS26T and Mu89 tumors.
- the administration of losartan alone did not affect the tumor growth rate (Figs. 11A and 11B).
- losartan significantly delayed the growth in both Mu89 and HSTS26T tumors-(Figs. 11A and 11B).
- the volume of HSTS26T tumors remained stable for up to 9 weeks in 50% of mice treated with losartan and HSV.
- mice treated with losartan and HSV had a delay in tumor growth.
- the growth delay in Mu89 tumors was only transient, 4 weeks after the virus injection all the tumors were 3-fold larger than the starting treatment size.
- mice with orthotopic pancreatic tumors were treated with DOXIL® and losartan.
- DOXIL® 4mg/kg, i.v.
- a sub- anti-tumor dose i.e., a dose that is not effective for treatment of cancer, e.g., a dose that is not effective to inhibit or prevent tumor growth and/or progression
- DOXIL® 4mg/kg, i.v.
- losartan or DOXIL alone did not affect the mean tumor weight (Fig. 11C).
- the tumors were significantly smaller (p ⁇ 0.001) than in mice that received DOXIL® alone (Figs. 11C and 11D).
- Example 6 The pattern of collagen distribution regulates the effectiveness of losartan
- Figs. 12A and 12B show striking differences between the collagen structure in Mu89 (Fig. 12A) and HSTS26T (Fig. 12B) tumors, respectively. Without wishing to be bound by theory, these differences in the collagen structure altered the virus propagation in these tumor types. In Mu89 tumors the collagen fiber network was well organized and formed finger-like projections into the tumor (Figs. 12 A and 13A).
- FIGs. 12B and 13B The dense collagen network seemed to slow down virus propagation but did not completely impede it, resulting in increased virus propagation and a more diffuse pattern of necrosis in this tumor (Fig. 14A).
- renin-angiotensin-aldosterone system 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 11 1 :2509- 2517; and Wolf G (2006) Kidney Int 70: 1914-1919).
- Angiotensin II has been reported to stimulate collagen production via both TGF- ⁇ dependent and independent pathways (Yang F, et al. (2009) Hypertension 54:877-884).
- Losartan and other RAAS inhibitors have reported to 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: 1 132-1 144), and reverse renal and cardiac 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, the inventors have demonstrated herein for the first time that losartan also inhibits collagen I production in tumors.
- an AHCM agent e.g., losartan
- the inventors also discovered that the organization of the collagen fibrillar network can affect nanoparticle distribution. This was striking because of significant differences in the structural organization of fibrillar collagen I between Mu89 and HSTS26T. In Mu89 tumors, thick bundles of fibrillar collagen I surround the tumor margins and form finger-like projections, which subdivide the tumor mass into isolated compartments and confine the viral infection to the injection site / isolated compartments (Figs. 12A and 13A).
- HSTS26T tumors have a mesh-like collagen structure, which hinders the virus spread but does not restrict viral particles to the injection site (Figs. 12B and 13B).
- the slower growth rate of HSTS26T than Mu89 tumors could also explain in part the enhanced efficacy of losartan combined with HSV in HSTS26T tumors.
- the collagen network organization plays an important role in limiting the penetration of large therapeutics in tumors.
- doses, administration methods and/or frequency of an AHCM agent e.g., losartan
- a cancer therapeutic e.g., HSV
- Pancreatic cancer patients treated with cytotoxic agents have a very high frequency of relapse with a 5 year survival of less than 5% (Li J, et al. (2010) AAPS J 12:223-232).
- the poor vascular supply and increased fibrotic content of pancreatic tumors most likely play a significant role in limiting the delivery and efficacy of cytotoxics (Olive KP, et al. (2009) Science 324: 1457-1461).
- the inventors show - in a mouse orthotopic model of human pancreatic cancer (L3.6pl) - that losartan increases both the intratumoral dispersion and extravascular penetration distance of i.v. injected nanoparticles.
- losartan are not limited to the interstitial space. Modifications to the RAAS system can also 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). Losartan-blockade of AGTRl 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.
- losartan did not affect tumor growth or the vascular density in HSTS26T tumors. Losartan can also reduce the proliferation of tumor cells expressing AGTRl (Rhodes DR, et al. (2009) Proc Natl Acad Set USA 106: 10284-10289). The inventors did not find a decrease in cancer cell proliferation (Fig. 15) or tumor size in the human melanoma Mu89, which express AGTRl (Fig. 16). The difference between their study and other prior studies might be due to differences in dosage.
- the dose of losartan was up to 15 fold higher than what was used in the inventors' study (Otake AH, et al. (2010) Cancer Chemother Pharmacol 66:79-87).
- the inventors have shown herein that a low dose of losartan that is ineffective for treatment of cancer by itself alone, can be used to improve the efficacy of a cancer therapy or an anti-cancer agent (even at a sub-therapuetic level) for treatment of cancer.
- the low dose of losartan can allow for a more clinically translatable protocol and avoid hypotensive complications.
- losartan administered at a low dose e.g., a dose not effective to reduce or prevent metastasis if administered alone
- an anti-metatstic agent e.g., at a dose less than what is typically administered by itself for treatment and/or prevention of metastasis
- losartan administered at a low dose e.g., a dose not effective to reduce or prevent metastasis if administered alone
- an anti-metatstic agent e.g., at a dose less than what is typically administered by itself for treatment and/or prevention of metastasis
- patients can be treated with a dose of 2mg/kg/day losartan, which is generally used for the treatment of patients with Marfan's syndrome (Brooke BS, et al. (2008) N Engl J Med 358:2787-2795).
- losartan and ARBs have limited side effects, losartan therapy is not recommended for patients with known renal disease.
- Losartan can induce renal insufficiency in patients with renal microvascular or macrovascular disease, or congestive heart failure (Sica DA, et al. (2005) Clin Pharmacokinet 44:797-814).
- Hyperkalemia can also occur in patients with poor renal function or patients who are concomitantly receiving potassium supplements or potassium sparing diuretics.
- angioedema caused by high levels of circulating angiotensin II can occur in patients treated with losartan (Sica DA, et al. (2005) Clin Pharmacokinet 44:797-814).
- Tumor drug resistance is generally believed to occur at many levels including increased drug efflux, drug inactivation, evasion from apoptosis, and alterations in target pathways (Longley DB, et al. (2005) J Pathol 205:275-292). Since losartan is not an antitumor agent, tumor resistance to losartan therapy after extended treatment can result from other mechanisms. Given that TGF-Bl activation is induced by different agents like MMPs and integrins in addition to TSP-1, tumor resistance to losartan could result from changes in TGF-Bl activation and signaling. However, long-term losartan therapy after myocardial infarction has been reported as not being associated with a reduction in antifibrotic properties (Schieffer B, et al. (1994) Circulation 89:2273-2282).
- losartan reduces the stromal collagen content in tumors and improves the penetration and therapeutic efficacy of nanoparticles (DOXIL ® , HSV) delivered both i.t. and i.v.
- Losartan also exhibits vasoactive and anti-metastatic properties that could increase its clinical application.
- losartan since losartan is already approved for clinical use, it represents a safe and effective adjunct for improving the efficacy of nanotherapeutics in cancer patients.
- CAFs isolated from human breast cancer biopsies were treated with losartan for 24 hrs prior to measurements of collagen and cytokine levels. Protein assays were done with commercial ELISA kits. All animal experiments were done with approval of the Institutional Animal Care and Use Committee. Losartan was administered i.p. at
- mice were treated with HSV (i.t.) and DOXIL ® (i.v. via tail vein) after 2 weeks of losartan treatment.
- Excised tumors were either snap frozen for biochemical analyses or fixed in paraformaldehyde, and embedded in paraffin or optimum cutting temperature compound (OCT) for immunohistochemistry.
- CAFs were isolated from human breast cancer biopsies using an art- recognized protocol, e.g., the protocol described in Orimo A, et al. (2005) Cell 121 :335-348.
- CAFs were plated in 24 well plates at a concentration of 500K cells/well. Cells were allowed 24 hrs to adhere to the plates before the addition of losartan at 10 ⁇ / ⁇ for 24 hrs (Schiittert JB, et al. (2003) Pflugers Arch 446:387-393). Treatment was done in low serum to reduce background collagen levels. Conditioned medium was collected at the end of the 24-hr treatment period and analyzed for collagen levels.
- TGF- ⁇ assays were performed with a human TGF- ⁇ ELISA kit (R&D Systems, Minneapolis, MN). The assay only measures the free-form of mature TGF- ⁇ 1. To measure total levels of TGF- ⁇ 1 the latent form of TGF- ⁇ 1 was activated with IN HC1. TSP-1 assays were performed with a human TSP- 1 ELISA kit (R&D Systems, Minneapolis, MN).
- Tumor sizes were monitored in spontaneous FVB/N- Tg(MMTV-PyVT) 634MU1/J mice and tumors selected for treatment when they reached a size of 4 to 6 mm in diameter (Guy CT et al. (1992) Mol Cell Biol 12:954-961).
- Cozaar (losartan potassium) tablets were ground 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 was administered by daily i.p. injections at a concentration of 10, 20 or 60 mg/kg/day for up to 2 weeks (Melo LG, et al. (1999) Am J Physiol 277:R624-R630). Tissue collection, embedding and staining
- Tumors for immuno staining analysis and quantification were harvested from mice, fixed in 4% paraformaldehyde, and embedded in paraffin or optimum cutting temperature compound (OCT) (Sakura Finetek Torrance, CA). OCT embedded tumors were soaked in sucrose solution for 24 hrs prior to embedding and freezing.
- OCT optimum cutting temperature compound
- the background- signal intensity for both collagen I and thrombospondin- 1 immunostaining was low and uniform.
- the inventors confirmed that the average signal intensity threshold lead to an accurate representation of the collagen and thrombospondin- 1 immunostaining and did not include the background signal.
- Second Harmonic Imaging (SHG) imaging was performed in 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 detected SHG signals at 405 nm was used.
- SCID mice bearing HSTS26T tumors in dorsal chambers were either treated with losartan (10, 20 or 60 mg/kg/day) or saline for the duration of the dose response experiment (15 days).
- Vascular markers were used to locate 4 regions of interest in each mouse and periodically returned to the same region of SHG imaging.
- SHG images were analyzed with a custom-built Matlab (The MathWorks, Inc., Natick, MA) code.
- the fraction of the region of interest (ROI) that was positive for the SHG signal was normalized to the amount of SHG signal obtained on day 1 of the dose response study (before initiation of losartan or saline treatment). Analysis of HSV infection and nanoparticle distribution
- Nanoparticles and oncolytic HSV were infused with a syringe pump (Harvard Apparatus Standard Pump 22, Holliston, Massachusetts) at a flow rate of 4 ⁇ / min.
- the inventors injected 10 ⁇ of HSV (2.5 x 10 5 t.u.) expressing the green fluorescent protein (GFP), or 10 ⁇ of fluorescent nanoparticles (diameter of 100 ⁇ ;
- the injected tumors were resected 30 min after the nanosphere injection and 24 hrs after the HSV infusion. Resected tumors were bisected at an angle perpendicular to the needle track, fixed in paraformaldehyde and frozen in OCT. All tumor sections were obtained perpendicular to the angle of the needle track. The entire tumor section was imaged with a confocal microscope (Olympus BX61WI) at 2x and images were reconstituted as mosaics.
- the nanosphere distribution and GFP -positive areas corresponds to the fraction of pixels brighter than the background signal.
- Intravenous injection A total volume of 10 ⁇ at a concentration of 3.6 x 10 13 nanoparticles / ml was injected via the tail vein. Twenty-four hrs later 50 ⁇ of FITC-lectin was injected to identify functional vessels. Five min after the lectin-injection tumors were resected, fixed in paraformaldehyde and embedded in OCT. Tumors were then sectioned before confocal imaging and analysis. The extent of nanosphere distribution was determined by measuring the fraction of pixels brighter than the background signal. Nanosphere penetration was determined by drawing contours around perfused vessels and recording the fraction of pixels positive for nanospheres in each contour. Contours extended out to 30 ⁇ for each perfused vessel. Using a previously described algorithm (Tong RT, et al. (2004) Cancer Res 64:3731-3736), the inventors fit the plot of nanosphere fraction and distance away from the vessel to an exponential and obtained a relative penetration depth of nanospheres from each vessel.
- mice with HSTS26T tumors implanted in a dorsal skin fold chamber were treated with i.p. injections of losartan (40mg/kg/day) for 1 week.
- Fluorescence recovery after photobleaching (FRAP) measurements were done with a custom built multiphoton microscope based on a previously described protocol (Chauhan VP, et al. (2009) Biophys J 97:330-336).
- IgG labeled fluorescein isothiocyanate (0.5 ml; 2mg/ml) was injected i.t. and used as the tracer.
- Ki67 staining was done on paraffin sections 21 days after HSV injection. Slides were microwave processed with Target Retrieval Solution (DAKO, Carpinteria, CA) prior to primary antibody detection. The entire tumor section was imaged at 2x
- mice Two weeks after the implantation of orthotopic pancreatic L3.6PL tumors, mice were randomly selected for losartan or saline treatment. A sub- anti-tumor dose of DOXIL ® (4 mg/kg) was infused i.v. via the tail vein after two weeks of losartan treatment (20 mg/kg/day). One week after the DOXIL injection, the tumors were resected and measured.
- DOXIL ® 4 mg/kg
- Scid mice bearing subcutaneous HSTS26T and MU89 tumors were randomly divided into control and losartan treated groups. Each arm (control and treated) was subsequently divided into HSV treated and non-HSV treated groups. Tumors that had reached 60 mm 3 after two weeks were selected for i.t. HSV injections. Tumors were treated with 10 ⁇ i.t. injections of either PBS or 2.5 x 10 5 transducing units (t.u.) of oncolytic HSV MGH2 (gift from E. Antonio Chiocca, Ohio State University, Columbus, OH). Two i.t. injections of oncolytic HSV separated by 24 hrs were administered.
- Example 7 Angiotensin blockade improves drug delivery by normalizing the tumor microenvironment
- angiotensin blockade “normalizes" interstitial matrix in solid tumors, including breast and pancreatic tumors (Fig. 17A).
- ARBs angiotensin receptor blockers
- ACE-Is angiotensin converting enzyme inhibitors
- the inventors also determined that ARBs and ACE-Is can decompress blood vessels to improve perfusion (Figs. 17B-17D), increase tumor hydraulic conductivity to repair vessel function (Fig.
- ARBs and ACE-Is can enhance the delivery of therapeutics, and thus have broad applicability for combination therapy with all classes of anti-cancer agents including small- molecule chemotherapeutics, biologies, and nanoparticle therapies.
- Angiotensin blockers offer numerous advantages over other approaches. Anti- angiogenic therapies normalize the vasculature alone and have been approved for only a limited number of indications. Meanwhile, ARBs and ACE-Is are FDA-approved as antihypertensives with manageable adverse effects. Matrix-degrading enzymes, which can normalize the collagen matrix, are not selective for tumors and can increase invasion and metastasis. ARBs and ACE-Is generally have no complications associated with matrix remodeling in normal tissues, leading to their safety as anti-hypertensives. ARBs and ACE- Is, as small-molecule agents, can also be delivered via nanovectors containing
- chemotherapeutics e.g., liposomes, nano-particles
- Anti- angiogenics the only FDA-approved adjuncts that enhance drug delivery to tumors, generally cannot improve delivery for larger particles as they can reduce the size of "pores" in vessel walls.
- angiotensin blockers presented herein can improve delivery for all classes of anti-tumor diagnostics and therapies.
- Example 8 In vitro screen to identify anti- hypertensive agents to lower collagen in solid tumors
- This Example provides an assay to rank anti-hypertensive (AH) agents based on their ability to lower collagen I level in tumors.
- the inventors determined that losartan reduced TGF- ⁇ activation and collagen I production in breast CAFs in vitro.
- Cells were treated with 10 ⁇ /L of losartan for 24 hrs.
- Losartan reduced by 90% the active-TGF- ⁇ levels (p ⁇ 0.05), while total TGF- ⁇ levels were unaffected.
- Anti-hypertensive agents Any FDA-approved angiotensin receptor blockers (ARBs) can be tested. Exemplary names and doses of these agents can be found via, but not limited to, http://www.globalrph.com/druglist.htm.
- angiotensin converting enzyme inhibitors also lower collagen, they do not target the receptor on cells and hence the inventors did not measure their effects on collagen I. Calcium channel blockers can also be evaluated for the collagen lowering effects.
- CAFs carcinoma-associated fibroblasts
- human cancer biopsies using a previously described protocol ( Orimo A, et al. (2005) Cell 121(3):335-348).
- CAFs should be plated in 24 well plates at a concentration of 500K cells/well and allowed 24 hrs to adhere to the plates before the addition of anti-hypertensive drug.
- all the losartan studies were performed at 10 ⁇ / ⁇ for 24 hrs, based on a published protocol (Schuttert JB, et al. (2003) Pflugers Arch 446(3):387-393).
- Treatment can be done in low serum to reduce background collagen levels.
- Conditioned medium can be collected at the end of the 24-hr treatment period and analyzed for total and activated TGF- ⁇ , TSP-1, CTGF and collagen levels.
- TGF- ⁇ assays were performed with a human TGF- ⁇ ELISA kit (R&D Systems, Minneapolis, MN). The assay only measures the free-form of mature TGF- ⁇ . To measure total levels of TGF- ⁇ the latent form of TGF- ⁇ was activated with IN HC1. TSP-1 assays were performed with a human TSP-1 ELISA kit (R&D Systems, Minneapolis, MN). CTGF ELISA kit can be purchased from Leinco (www.leinco.com).
- CAFs carcinoma associated fibroblasts
- Example 9 Combination of angiotensin blockade with inhibition of alternate profibrotic pathways to improve drug delivery to tumors
- angiotensin signaling blockade improves drug delivery, at least partly, through two mechanisms: it relaxes the inherent compressive force in tumors to improve vessel perfusion, and it reduces the viscoelastic and steric hindrance on drug transport directly imparted by the matrix.
- Angiotensin signaling blockade can safely inhibit activation of the profibrotic TGF- beta and CTGF pathways downstream to produce these changes.
- endothelin receptor blockers (ERBs) and PDGF inhibitors (PDGF-Is) can be used in combination with angiotensin blockers.
- ERBs treat pulmonary arterial hypertension and can be used as a class of therapy for cancer (Nelson et al. (2003) Nature Reviews Vol. 3 : 110- 116), for example with angiotensin blockers.
- PDGF-Is haven been reported for their potential anti-vascular effects in tumors (Baluk et al. (2005) Current Opinion in Genetics & Development 15: 102- 11 1, Andrae et al. (2008) Genes & Development 22: 1276- 1312). Endothelin blockade has been reported to reduce fibrogenesis in the 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 through inhibition of TGF-beta synthesis (Ogata et al. (2002) Clinical Science 103 (Suppl.
- an angiotensin blockade with endothelin- 1 and/or PDGF blockade - with careful dosing - should produce an additive improvement to drug delivery with minimal additional toxicity.
- endothelin-1 and/or PDGF blockade can be used at a sub-therapuetic dose in combination with an angiotensin blockade, which can be used at a sub-anti- hypertensive dose and/or sub- anti-tumor dose, for improved drug delivery and/or treatment of cancer.
- Example 10 Angiotensin inhibitors decompress tumor vessels to enhance drug delivery
- a specific anti-VEGF inhibitor (the antibody bevacizumab) showed no overall survival benefit when added to chemotherapy in randomized, double-blind, phase-Ill trials for breast, pancreatic, kidney and non-small cell lung cancers (Escudier B, et al, Journal of Clinical Oncology. 2010; 28(13): 2144-50;
- angiotensin inhibition can also modulate the non- vascular tumor microenvironment (Table 1). Therefore, angiotensin inhibitors could synergistically enhance chemotherapy by increasing drug and oxygen delivery to the tumor. Chemotherapy effectiveness is dependent on drug delivery (Jain RK. Science. 2005; 307(5706): 58-62). Furthermore, inadequate oxygen delivery can lead to hypoxia-induced drug resistance (Wilson WR, et al, Nat Rev Cancer. 2011; 11(6): 393-410), selection for aggressive cells (Harris AL. Nature reviews. 2002; 2(1): 38-47), and immune suppression in tumors
- mice were injected with lectin as a marker for perfused (patent and functional) vessels before tumor excision, and then immunostained tissue sections with an anti-CD31 antibody, which marks both perfused and non-perfused vessels (Olive KP, et al, Science. 2009; 324(5933): 1457-61). These tumors were found to be severely hypo-perfused (Fig. 21): only 23% of vessels in E0771 breast tumors and 21% in AK4.4 pancreatic tumors were perfused with blood (Figs. 17D and 22A).
- a 40mg/kg dose of losartan which does not decrease blood pressure in tumor-bearing mice (Fig. 23), significantly improved the perfused vessel fraction to 43% in E0771 and 45% in AK4.4 (Figs. 17D and 22A).
- Losartan treatment increased vessel diameters in E0771 (Fig. 22B), suggesting vascular decompression as the mechanism of action.
- losartan did not affect the CD31+ vessel density in these tumors (Figs. 22C-22D).
- losartan can increase blood supply in tumors by opening existing collapsed blood vessels.
- Collagen levels were used as a metric of matrix production as described in Diop-Frimpong et al, Proc Natl Acad Sci USA. 2011; 108(7): 2909-14, and losartan was found to reduce the collagen I levels in E0771 and AK4.4 tumors (Fig. 24A and 24C). Moreover, losartan decreased collagen I concentration, measured based on staining intensity, in E0771 and AK4.4 tumors (Figs. 24B-24C). Of note, dense collagen seemed to colocalize with low- perfusion areas (Fig. 24C). Next, solid stress in these tumors was measured using recently established technique described in Stylianopoulos T, et al, Growth-induced mechanical stress in murine and human tumors: causes, consequences and remedies, under review.
- Nanoparticle penetration rates were quantified as transvascular mass flux per unit vascular surface area and transvascular concentration difference, termed as the effective permeability. It was found that losartan improved effective permeability for 12nm, 60nm, and 125nm nanoparticles (Fig. 29E), i.e., for the entire size range of nanomedicines in clinical use.
- microenvironmental normalization with ARBs and ACE-Is increases the delivery of oxygen and all sizes of therapeutics to tumors.
- angiotensin inhibitors could act as adjunct therapies to synergistically improve the effectiveness of small-molecule chemotherapeutics.
- losartan was tested in combination with doxorubicin in E0771 and 4T1 tumors, or with 5FU in AK4.4 tumors. It was found that whereas losartan or doxorubicin given alone had no significant effect on tumor growth rate, the combination significantly delayed E0771 and 4T1 tumor growth (Figs. 19A-19B and 35).
- losartan alone despite increasing blood perfusion in tumors, did not shorten survival in mice bearing E0771, 4T1 or AK4.4 tumors (Figs. 19C, 34-35).
- losartan monotherapy did not increase metastasis in AK4.4, and its combination with 5-FU appeared to reduce the incidence and size of metastases (Table 2).
- angiotensin inhibitors can improve the effectiveness of small molecule chemotherapeutics through anti-matrix effects.
- anti-VEGF therapies shrink vessel pores (Chauhan VP, et al, Nature Nanotechnology. 2012; advance online publication), reduce interstitial fluid pressure (IFP) (Tong RT, et al, Cancer research. 2004; 64(1 1): 3731-6); Goel S, et al, Physiol Rev. 2011; 91(3): 1071- 121) and increase perfusion in patients (Sorensen AG, et al, Cancer research. 2012; 72(2): 402-7).
- IFP interstitial fluid pressure
- vascular normalization is suited to well-perfused tumors (Sorensen AG, et al, and Goel S, et al), but may not work in desmoplastic tumors such as in pancreatic cancer when a significant fraction of tumor vessels are collapsed.
- most anti- angiogenic drugs lead to hypertension in a significant number of cancer patients and this hypertension is currently managed with a variety of anti-hypertensive drugs (Keizman D, et al, Eur J Cancer. 201 1; 47(13): 1955-61.).
- Another example is anti-hyaluronan enzymatic therapy, which can also increase vessel diameter in pancreatic tumors
- ARBs and ACE-ls would be compatible with all these strategies, due to the differences in their targets and mechanisms. Moreover, in contrast to the other drugs and agents that can improve drug delivery, ARBs and ACE-ls are the only FDA-approved drugs that can reduce solid stress thus far. Despite this great promise, ARBs and ACE-ls may be limited in usage by several factors. Notably, the activity of these drugs should be most effective in desmoplastic tumors, such as in breast, stomach, and pancreatic cancer. In addition, not all breast tumors are desmoplastic, which could explain how the disease is sometimes curable with chemotherapy, and thus not all patients may be candidates for ARBs and ACE-ls. Additionally, these agents will only work in patients with desmoplastic tumors, such as in breast, stomach, and pancreatic cancer. In addition, not all breast tumors are desmoplastic, which could explain how the disease is sometimes curable with chemotherapy, and thus not all patients may be candidates for ARBs and ACE-ls. Additionally, these agents will only work
- angiotensin-II receptor type-1 positive tumors Furthermore, ARBs and ACE-ls would be contraindicated for patients with low blood pressure or certain other co-morbidities.
- ARBs and ACE-ls were found to increase the delivery of oxygen, small-molecule chemotherapeutics, and nanomedicine through
- Angiotensin inhibitors (losartan, lisinopril, valsartan, and candesartan) were obtained as pills. The pills were crushed using a mortar and pestle and the powder was dissolved in phosphate buffered saline (PBS) over 24 hours. The solutions were then sterile filtered for injection. Doxorubicin and 5-FU were obtained as solutions for injection, and were injected without modification. All drugs were purchased from the pharmacy at Massachusetts General Hospital.
- PBS phosphate buffered saline
- AK4.4 was kindly provided by Dr. Nabeel Bardeesy, and was isolated from mice generating spontaneous pancreatic tumors (Kras° 12 and p53 +/" ).
- Orthotopic pancreatic tumors were generated by implanting a small piece (1mm 3 ) of viable tumor tissue (from a source tumor in a separate animal) into the pancreas of a male FVB mouse (AK4.4 model) or C57BL/6 (Pan-02 model) mouse.
- Orthotopic breast tumors were similarly generated by implanting a chunk of viable tumor tissue into the mammary fat pad of a female severe combined immunodeficient (SCID) mouse. All animal procedures were carried out following the Public Health Service Policy on Humane Care of Laboratory Animals and approved by the Institutional Animal Care and Use Committee of Massachusetts General Hospital.
- mice bearing orthotopic E0771 were split into treatment groups, time-matched for time after implantation and size-matched for tumor volume at this time ( ⁇ 100mm 3 ).
- mice bearing orthotopic AK4.4 were split into treatment groups, size- matched for tumor volume ( ⁇ 22mm 3 ), 6 days after implantation. The mice were then treated with 40mg/kg losartan or an equal volume of PBS intraperitoneally each day for 6 (E0771) or 7 (AK4.4) days.
- mice were slowly ( ⁇ 2min) injected with ⁇ of lmg/mL biotinylated lectin (Vector Labs), administered retro- orbitally 5min prior to tumor removal.
- the mice were also injected with 60mg/kg of lOmg/mL pimonidazole lhr prior to tumor removal.
- the tumors were then excised, fixed in 4% formaldehyde in PBS (30min/mm diameter of tissue), incubated in 30% sucrose in PBS overnight at 4°C, and frozen in optimal cutting temperature compound (Tissue-Tek).
- Transverse tumors sections 40 ⁇ thick, were immunostained with antibodies to endothelial marker CD31, and counterstained by mounting with DAPI- containing medium (Vector Labs).
- DAPI- containing medium Vector Labs
- collagen I was detected using the LF-67 antibody provided by Dr. Larry Fisher (National Institute of Dental Research, Bethesda, MD).
- Solid stress was measured using the tumor opening technique as described previously (Stylianopoulos T, et al, Growth-induced mechanical stress in murine and human tumors: causes, consequences and remedies, under review. 2012). When the tumors reached a size of ⁇ lcm in diameter, the mice were anesthetized.
- each tumor was excised, washed with Hanks' Balanced Salt Solution (HBSS) and its three dimensions were measured.
- HBSS Hanks' Balanced Salt Solution
- Each tumor was cut along its longest axis, to a depth of 80% of its shortest dimension, using a scalpel.
- the tumors were allowed to relax for 10 minutes in HBSS to diminish any transient, poro-elastic responses.
- the opening resulting from the cut was measured at the middle of the cut at the surface of the tumor. Solid stress is proportional to the size of the opening relative to the size of the dimension perpendicular to the cut.
- mice bearing orthotopic AK4.4 were split into treatment groups, size-matched for tumor volume ( ⁇ 22mm 3 ), 6 days after implantation. The mice were then treated with 40mg/kg losartan or an equal volume of PBS intraperitoneally each day for 7 days. On the day of the last treatment, mice were injected with lOOmg/kg 5-FU, administered retro-orbitally 30min prior to tumor and organ removal. The tissue was dabbed of excess blood then snap-frozen in liquid nitrogen for analysis. 5-FU was isolated from the tissues and measured using liquid-liquid extraction followed by reverse- phase high-performance liquid chromatography with tandem mass- spectrometry.
- Tissue oxygenation p02 in the tumors was measured using
- Oxyphor R2 Oxyphor R2 (Oxygen Enterprises) was injected retro-orbitally 12hrs prior to imaging, and was reinjected immediately prior to imaging along with 2MDa FITC-dextran (Sigma- Aldrich) for functional vascular tracing. A mosaic image of the tumor was collected, and oxygen was then measured in an evenly spaced 12x12 grid at four depths (60, 120, 180, and 240 ⁇ ) in the tumor.
- the phosphorescence lifetime of the probe was measured after each of several repeated brief intense pulses of 1020nm laser light, and these lifetime measurements were combined.
- a two-component model was used to calculate the oxygen tension from each lifetime measurement, accounting for binding and quenching of the probe by both oxygen and proteins.
- the grid was then overlayed on the mosaic images to make an oxygen map.
- Nanoparticle penetration A mixture of nanoparticles with diameters of
- nanoparticle samples under 800nm multiphoton excitation were prepared under 800nm multiphoton excitation. Following retro-orbital injection of 200 ⁇ . with these concentrations, multiphoton imaging was carried out as described above at depths from 0 - 20 ⁇ ⁇ , with 2.76 ⁇ steps and 2.76 ⁇ 2.76 ⁇ pixels. Images were taken every 3min at each region of interest for a duration of lhr. Images were analyzed using custom analysis software developed in Matlab (The Mathworks) as described previously (Chauhan VP, et al, Nature Nanotechnology. 2012; advance online publication; Chauhan VP, et al, Angewandte Chemie International Edition. 2011; 50(48): 11417-20).
- the analysis approach involved 3D vessel tracing to create vessel metrics and a 3D map of voxel intensity versus distance to the nearest vessel over time. Images were also corrected for sample movement over time with 3D image registration.
- the effective permeability was where J t is the transvascular flux, S v is the vessel surface area, C v is the concentration of the probe in the vessel, C is the concentration of the probe immediately extravascular, P eff is the effective permeability (Chauhan VP, et al, Nature Nanotechnology. 2012; advance online publication; Chauhan VP, et al, Angewandte Chemie International Edition.
- t time after the initial image
- r is the distance from the vessel central axis
- R is the vessel radius at that point along the vessel. Fluorescence intensities were used as these concentrations. The calculation was made as an average over the entire imaged volume for each tumor.
- Pancreatic tumor growth and metastasis studies Mice bearing orthotopic AK4.4 pancreatic tumors were split into treatment groups, size-matched for tumor volume (22mm 3 ), 6 days after implantation. The mice were treated with 40mg/kg losartan or an equal volume of PBS intraperitoneally on day 7 after implantation and each subsequent day. The mice were then treated with either 60mg/kg 5-FU or an equal volume of saline by intravenous injection on days 9 and 13 after implantation. Tumors were extracted on day 14 for measurement using calipers. Tumor growth was quantified using the size at day 14. For metastasis studies, mice were treated with losartan or PBS on day 1 1 after implantation, then with 5-FU or saline on days 13 and 17. Metastatic burden was assessed at death.
- Mean arterial blood pressure Mice bearing orthotopic AK4.4 pancreatic tumors were used for blood pressure measurements. Mean arterial blood pressure was measured by cannulation of the left carotid artery after a longitudinal skin incision above the trachea, as described previously (Zlotecki RA, et al, Microvasc Res. 1995; 50(3): 429-43). After removal of the submandibular gland, the paratracheal muscles was split and the left carotid artery was isolated. The cranial end of the artery was ligated with a 6- 0 silk suture and another suture was tied loosely around the central part of the artery. A metal clamp was then positioned caudally to stop blood flow during the cannulation.
- a polyethylene catheter (PE- 10, Becton-Dickinson) filled with heparinised saline was then be inserted through a hole cut proximally to the cranial ligature, and the other suture was tied tightly around the tubing and artery. The clamp was then removed and the end of the tubing was connected to a pressure transducer for the measurement of blood pressure.
- Interstitial hydraulic conductivity was measured as described previously (Mok W, et al, Cancer Res. 2007; 67(22): 10664-8; Wabb EA, et al, Cancer research. 197 '4; 34(10): 2814-22).
- the tumors were excised, and a 3mm biopsy punch was used to cut a cylindrical tissue block from each.
- a scalpel was then used to cut a 1.7mm-thick disc of viable tumor tissue from this cylindrical block.
- the disc-shaped tissue block was then placed into a clamp with a fluid flow channel.
- a pressure head of 10cmH 2 O was applied, and a small bubble was created to measure the fluid velocity in the 0.58mm diameter tubing connecting the pressure head to the clamp. Measurements were taken over 5- lOmin per tumor.
- Example 11 Losartan pharmacokinetic/pharmacodynamic (PK/PD) analysis in mouse pancreatic tumor model
- This example presents a study that compared the pump administration and pulsatile injections of losartan based on the results of PK/PD analysis in mouse AK4.4 pancreatic tumor model.
- PBS Phosphate buffered saline
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US11433136B2 (en) | 2015-12-18 | 2022-09-06 | The General Hospital Corporation | Polyacetal polymers, conjugates, particles and uses thereof |
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US11155674B2 (en) | 2017-01-20 | 2021-10-26 | Massachusetts Institute Of Technology | Polymerizable sulfonamide compounds and polymers thereof |
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US10988491B2 (en) | 2018-08-17 | 2021-04-27 | Massachusetts Institute Of Technology | Degradable polymers of a cyclic silyl ether and uses thereof |
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