Methods for the treatment of leptomeningeal carcinomatosis

Abstract

The present invention features a method of treating leptomeningeal carcinomatosis in a subject using a peptide-therapeutic conjugate as exemplified by the agent ANG1005.

Description

BACKGROUND

Leptomeningeal carcinomatosis (LC) is a rare complication of cancer in which the disease spreads to the membranes (meninges) surrounding the brain and spinal cord. LC occurs in approximately 5% of people with cancer and is usually terminal. If left untreated, median survival is 4-6 weeks; if treated, median survival is 2-3 months. LC may occur at any stage of cancer, either as a presenting sign or as a late complication, although it is associated frequently with relapse of cancer elsewhere in the body.

LC is generally considered incurable and is difficult to treat. Treatment goals generally include improvement or stabilization of the patients neurologic status, prolongation of survival, and palliation. Thus, there is a need for therapeutics and therapeutic regimens capable of treating LC.

SUMMARY OF THE INVENTION

The present invention is based on the discovery that ANG1005, a conjugate of Angiopep-2 and three molecules of paclitaxel, was successful in treating patients with leptomeningeal carcinomatosis (LC). This conjugate is able to treat patients having LC, even where the patient is not responsive to standard chemotherapeutic agents. Since cancer cells in LC are located in the CSF and/or in the meninges, to treat LC, a compound likely needs to be transported across the blood-CSF barrier, which, unlike the blood-brain barrier which is mainly comprised of endothelial cells with tight junctions, is mainly comprised of epithelial cells. It has recently been shown that the cells of the blood-CSF barrier express LRP receptors (see Fujiyoshi et al. Journal of Neurochemistry, 2011, 118:407-415). As Angiopep-2 is known to interact with the LRP receptor, it is likely able to cross the blood-CSF barrier by LRP-mediated transcytosis, thereby transporting paclitaxel to the cancer cells in the CSF and/or in the meninges.

Accordingly, the invention features a method for the treatment of leptomeningeal carcinomatosis including administering to a subject in need thereof (e.g., a subject identified as having leptomeningeal carcinomatosis or suspected of having leptomeningeal carcinomatosis, e.g., based on radiological, neurological and/or cytological evaluations) an effective amount of a compound or pharmaceutically acceptable salt thereof including: (a) a polypeptide including the sequence of Angiopep-1 (TFFYGGCRGKRNNFKTEEY, SEQ ID NO: 1), Angiopep-2 (TFFYGGSRGKRNNFKTEEY SEQ ID NO: 2), or Angiopep-2-4D (TFFYGGS(D-R)G(D-K)(D-R)NNF(D-K)TEEY, SEQ ID NO: 3, wherein D-R refers to D-Arginine and D-K refers to D-lysine); and (b) an anticancer agent (e.g., paclitaxel), wherein the anticancer agent is conjugated to the polypeptide. In some embodiments, the compound includes a polypeptide having the sequence of Angiopep-2. In some embodiments, the anticancer agent is a taxane such as paclitaxel or docetaxel, vinblastine, vincristine, etoposide, doxorubicin, cyclophosphamide, melphalan, or chlorambucil. In some embodiments, the anticancer agent is conjugated to the polypeptide by a linker. In some embodiments, the linker has the structure:

In some embodiments, the anticancer agent is conjugated to the polypeptide through the primary amine at the N-terminus, the primary amine of the lysine at position 10, and/or the lysine at position 15. In some embodiments, the compound has the structure:

In some embodiments, the compound has the structure:

In some embodiments, the compound has the structure:

In some embodiments, the primary origin of the leptomeningeal carcinomatosis is a solid tumor (e.g., a brain tumor such a glioblastoma or medullablastoma, a breast tumor (e.g., a breast tumor identified as a HER2 positive tumor, a breast tumor identified as a HER2 negative tumor, breast tumor identified as ER positive, a breast tumor identified as ER negative, a breast tumor identified as PR positive, a breast tumor identified as PR negative, or a breast tumor identified as a triple negative breast tumor), a lung tumor such as a non-small cell lung tumor or small cell lung tumor, a gastrointestinal tumor, or malignant melanoma. In some embodiments, the primary origin of the LC is a liquid tumor (e.g., leukemia or lymphoma). In some embodiments, the primary origin of the leptomeningeal carcinomatosis is unknown (also known as cancer of unknown primary origin or occult cancer). In some embodiments, the tumor includes cells that express efflux pumps such as MDR1. In some embodiments, the primary origin of the leptomeningeal carcinomatosis is a breast tumor, e.g., a breast tumor identified as a HER2 positive tumor, a breast tumor identified as a HER2 negative tumor, breast tumor identified as ER positive, a breast tumor identified as ER negative, a breast tumor identified as PR positive, a breast tumor identified as PR negative, a breast tumor identified as HER2 positive, ER positive, and PR positive, a breast tumor identified as HER2 positive, ER positive, and PR negative, a breast tumor identified as HER2 positive, ER negative, and PR positive, a breast tumor identified as HER2 positive, ER negative, and PR negative, a breast tumor identified as ER positive, HER2 negative, and PR negative, a breast tumor identified as ER positive, HER2 negative, and PR positive, a breast tumor identified as PR positive, HER2 negative, and ER negative, or a breast tumor identified as a triple negative breast tumor. In some embodiments, the primary origin of the leptomeningeal carcinomatosis is a breast tumor, e.g., a breast tumor identified as a HER2 positive tumor, a breast tumor identified as a HER2 negative tumor, or a breast tumor identified as a triple negative breast tumor.

In some embodiments, the primary cancer has also metastasized to the liver, lungs, brain, bone, the lining of the abdomen or pelvis (peritoneum), organs of the abdomen such as the bowel, bladder, or uterus. In some embodiments, the primary cancer is in the lymph system. In some embodiments, the subject has at least one metastasis outside the brain, lung, liver, kidney, or eye.

In some embodiments, the subject previously received another anticancer therapy (e.g., an anticancer therapy including a chemotherapeutic agent such as a taxane, a platinum-based agent, an anthracycline, an anthraquinone, an alkylating agent, a HER2 targeting therapy (e.g., a HER2 antibody), vinorelbine, a nucleoside analog, ixabepilone, eribulin, cytarabine, a hormonal therapy, methotrexate, capecitabine, lapatinib, 5-FU, vincristine, etoposide, or any combination thereof). In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after the previously received anticancer therapy. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with a taxane, e.g., paclitaxel or docetaxel. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with methotrexate. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with a HER2 targeting therapy. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with a platinum-based agent. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with an anthracycline. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with an anthraquinone. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with an alkylating agent. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with vinorelbine. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with a nucleoside analog. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with ixabepilone. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with eribulin. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with cytarabine. In some embodiments, the primary cancer and/or LC failed to respond to the previously received anticancer therapy and/or relapsed after treatment with a hormonal therapy.

In some embodiments, the primary cancer and/or LC may be drug resistant (e.g., the cancer includes cells that do not respond to treatment with one or more anticancer agents) or include drug resistant cells (e.g., cells that expresses MDR1). The primary cancer and/or LC may be or may include cells that are resistant to any chemotherapeutic agent including paclitaxel, carboplatin, cisplatin, doxorubicin, topotecan, gemcitabine, docetaxel, a taxane derivative, or any agent described herein. In some embodiments, the primary cancer and/or LC is resistant to the previously received anticancer therapy.

In some embodiments, the method further includes the step of administering an additional anticancer therapy (e.g., an anticancer therapy including radiation therapy such as whole brain radiation therapy or stereotactic radiosurgery and/or a chemotherapeutic agent such as a taxane, a platinum-based agent, an anthracycline, an anthraquinone, an alkylating agent, a HER2 targeting therapy, vinorelbine, a nucleoside analog, ixabepilone, eribulin, cytarabine, a hormonal therapy, a bisphosphonate, methotrexate, capecitabine, lapatinib, 5-FU, vincristine, or etoposide). In some embodiments, the method further includes administration of methotrexate, an alkylating agent, cytarabine, or a HER2 antibody. In some embodiments, the method further includes administration of radiation therapy. In some embodiments, the additional anticancer therapy is administered before a compound of the invention. In some embodiments, the additional anticancer therapy is administered after a compound of the invention. In some embodiments, the additional anticancer therapy is administered simultaneously with a compound of the invention. In some embodiments, the method further includes the step of administering a palliative therapy, e.g., an analgesic, an anticonvulsant, an antidepressant, an anxiolytic, a psychostimulant, modafinil, palliative radiation, corticosteroids, an H1 antagonist, a hematopoietic growth factor, and/or a blood transfusion.

In some embodiments, the method includes administering a compound of the invention in a pharmaceutical composition. In some embodiments, the pharmaceutical composition includes: (a) a compound of the invention (e.g., ANG1005); (b) an optional tonicity agent; (c) a buffering agent (e.g., a buffering agent that maintains a pH of 4.5-6); (d) a bulking agent; (e) a solubilizing agent (e.g., a solubilizing agent that is not ethoxylated castor oil); and (f) 0.2 to 10% DMSO. In some embodiments, the tonicity agent is sodium chloride. In some embodiments, the buffering agent is glycine, citric acid, or lactic acid. In some embodiments, the bulking agent is mannitol or sorbitol. In some embodiments, the solubilizing agent is polyoxyethylene ester of a fatty acid (e.g., 12-Hydroxystearic acid-polyethylene glycol copolymer). In some embodiments, the composition is substantially free from ethoxylated castor oil or is free of ethoxylated castor oil. In some embodiments, the composition is dissolved in water. In some embodiments, the composition includes:

Compound                         Percentage (by non-water weight)
ANG1005                          1.8-2.3%
Tonicity agent                   9-11%
Buffer (e.g., lactic acid or citric acid) 4.5-6%
Bulking agent (e.g., mannitol)   8-10%
12-Hydroxystearic acid-polyethylene 69-75%
glycol copolymer
DMSO                             0.2-2%

In some embodiments, the composition includes:

Compound                         Percentage (by non-water weight)
ANG1005                          about 2%
Tonicity agent                   about 10%
Buffer (e.g., lactic acid or citric acid) about 5%
Bulking agent (e.g., mannitol)   about 9%
12-Hydroxystearic acid-polyethylene about 72%
glycol copolymer
DMSO                             about 1%

The compound or composition of the invention may be administered in a dosage of about 1, 10, 25, 50, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000, 2500, or 3000 mg/m², or any range between these numbers. In some embodiments, the dosage is between 100 mg/m² and 2000 mg/m² or between 300 mg/m² and 1000 mg/m². In some embodiments, the dosage is between 300 and 650 mg/m² (e.g., 550-625 mg/m²). In some embodiments the dosage is between 400 and 650 mg/m². In still further embodiments the dosage is between 400 and 600 mg/m² (e.g. 400, 470, 550 or 600 mg/m²). The compound of the invention may be administered by any means known in the art, e.g., intravenously, orally, intraarterially, intranasally, intraperitoneally, intramuscularly, subcutaneously, transdermally, or per os to the patient. In some embodiments, the compound is administered intravenously. In some embodiments, the compound is not administered intrathecally. In some embodiments, the compound of the invention is administered weekly (i.e., about every seven days). In some embodiments, the compound of the invention is administered bi-weekly (i.e., about every fourteen days). In some embodiments, the compound of the invention is administered tri-weekly (i.e., about every twenty one days). In some embodiments, the compound of the invention is administered at an interval of greater than twenty one days.

In some embodiments, at least one neurological symptom (e.g., headaches, gait difficulties, memory problems, incontinence, sensory abnormalities, or any neurological symptom described herein) of the subject is partially or completely alleviated, ameliorated, relieved, inhibited, delayed, or is reduced in severity after administration of a compound of the invention. In some embodiments, at least one lesion or leptomeningeal metastasis is decreased in size in the subject after administration of a compound of the invention. In some embodiments, the amount of cancer cells in the CSF is decreased in the subject after administration of a compound of the invention. In some embodiments, the flow of CSF in the subject is increased after administration of a compound of the invention.

Definitions

As used herein, the term “administration” refers to the administration of a composition (e.g., a compound, a conjugate, or a preparation that includes a compound or conjugate as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal.

A cancer “determined to be drug resistant,” as used herein, refers to a cancer that is drug resistant, based on unresponsiveness or decreased responsiveness to a chemotherapeutic agent, or is predicted to be drug resistant based on a prognostic assay (e.g., a gene expression assay).

By a “drug resistant” cancer is meant a cancer that does not respond, exhibits a decreased response to, or is predicted to be drug resistant based on a prognostic assay (e.g., a gene expression assay) one or more chemotherapeutic agents (e.g., any agent described herein).

The term “effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and/or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and/or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence and/or severity of, and/or delays onset of, one or more symptoms of the disease, disorder, and/or condition. Those of ordinary skill in the art will appreciate that the term “effective amount” does not in fact require successful treatment be achieved in a particular individual. Rather, an effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that particular subjects may, in fact, be “refractory” to an “effective amount.” To give but one example, a refractory subject may have a low bioavailability such that clinical efficacy is not obtainable. In some embodiments, reference to an effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweart, tears, urine, etc). Those of ordinary skill in the art will appreciate that, in some embodiments, an effective amount may be formulated and/or administered in a single dose. In some embodiments, an effective amount may be formulated and/or administered in a plurality of doses, for example, as part of a dosing regimen.

An “ER positive tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that has ER receptors on the surface of the cancer cells in the tumor and/or a tumor that expresses the gene for ER. An “ER negative tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that does not have ER receptors on the surface of the cancer cells in the tumor, a tumor that has a number of ER receptors below a predetermined level, or a tumor that does not express the gene for ER. The ER status of a tumor may be determined using methods known in the art, e.g., tests performed on biopsy samples such as immunohistochemistry or fluorescence in situ hybridization or by measurement of serum ER by ELISA.

As used herein, the term “failed to respond to a prior therapy” or “refractory to a prior therapy,” refers to a cancer or LC that progressed despite treatment with the therapy.

A “HER2 positive tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that has HER2 receptors on the surface of the cancer cells in the tumor and/or a tumor that expresses the gene for HER2. A “HER2 negative tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that does not have HER2 receptors on the surface of the cancer cells in the tumor, a tumor that has a number of HER2 receptors below a predetermined level, or a tumor that does not express the gene for HER2. The HER2 status of a tumor may be determined using methods known in the art, e.g., tests performed on biopsy samples such as immunohistochemistry or fluorescence in situ hybridization or by measurement of serum HER2 by ELISA.

A “palliative therapy,” as used herein refers to an therapy administered to a subject for the purpose of improving quality of life, e.g., by relieving one or more symptoms or side effects associated with a disease.

As used herein, the term “pharmaceutical composition” refers to an active compound, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, active compound is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.

A “pharmaceutically acceptable excipient,” as used herein, refers any inactive ingredient (for example, a vehicle capable of suspending or dissolving the active compound) having the properties of being nontoxic and non-inflammatory in a subject. Typical excipients include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those of ordinary skill in the art are familiar with a variety of agents and materials useful as excipients.

The term “pharmaceutically acceptable salt,” as use herein, refers to those salts of the compounds described here that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P. H. Stahl and C. G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable organic acid.

The compounds of the invention may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art.

Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

A “PR positive tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that has PR receptors on the surface of the cancer cells in the tumor and/or a tumor that expresses the gene for PR. A “PR negative tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that does not have PR receptors on the surface of the cancer cells in the tumor, a tumor that has a number of PR receptors below a predetermined level, or a tumor that does not express the gene for PR. The PR status of a tumor may be determined using methods known in the art, e.g., tests performed on biopsy samples such as immunohistochemistry or fluorescence in situ hybridization or by measurement of serum PR by ELISA.

The term “primary origin,” as used herein, refers to the organ in the body of the subject where the cancer began (e.g., the breast, lung, skin, gastrointestinal tract). The primary origin of a cancer may be identified using methods known in the art, e.g., medical imaging, examination of biopsy samples with immunohistochemistry techniques, and/or gene expression profiling.

The term “subject,” as used herein, refers to a human or non-human animal (e.g., a mammal such as a non-human primate, horse, cow, or dog).

The term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

A “therapeutic regimen” refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome.

The term “treatment” (also “treat” or “treating”), in its broadest sense, refers to any administration of a substance (e.g., provided compositions) that partially or completely alleviates, ameliorates, relives, inhibits, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms, features, and/or causes of a particular disease, disorder, and/or condition. In some embodiments, such treatment may be administered to a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition. Alternatively or additionally, in some embodiments, treatment may be administered to a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition. In some embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and/or condition. In some embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and/or condition.

A “triple negative tumor,” as used herein, refers to a tumor, e.g., a breast tumor, that does not have estrogen receptors, progesterone receptors, or human epidermal growth factor receptor 2 on the surface of the cancer cells in the tumor and/or does not substantially express the genes for estrogen receptor, progesterone receptor, or HER2. The ER, PR, and HER2 status of a tumor may be determined using methods known in the art, e.g., tests performed on biopsy samples such as immunohistochemistry or fluorescence in situ hybridization or by measurement of serum ER, PR, and/or HER2 by ELISA.

Other features and advantages of the invention will be apparent from the following Detailed Description, the drawings, and the claims.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is an image illustrating accumulation of an Angiopep-Cy5.5 conjugate in the meninges and parenchyma of a living mouse brain 24 hours post iv administration.

FIG. 2 is an image of a CT scan of a patient's brain prior to (FIG. 2A) and following (FIG. 2B) treatment with ANG1005.

FIG. 3 is a waterfall plot of the intracranial responses to ANG1005 of patients with leptomeningeal carcinomatosis.

FIG. 4 is a graph of Kaplan-Meier estimates for the survival of patients with leptomeningeal cacrinomatosis treated with ANG1005.

DETAILED DESCRIPTION

The present inventors have discovered that administration of a peptide-drug conjugate (e.g., ANG1005) is capable of treating leptomeningeal carcinomatosis (LC) in a subject. As LC is generally considered incurable, there is a need for therapeutics and therapeutic regimens capable of treating LC.

Leptomeningeal Carcinomatosis

The methods of the invention include treatment of a subject having LC. LC (also known as leptomeningeal metastasis or leptomeningeal disease) is a rare complication of cancer in which the disease spreads to the membranes (meninges) surrounding the brain and spinal cord. LC occurs in approximately 5% of people with cancer and is usually terminal. If left untreated, median survival is 4-6 weeks; if treated, median survival is 2-3 months. LC may occur at any stage of cancer, either as a presenting sign or as a late complication, though it is associated frequently with relapse of cancer elsewhere in the body.

LC occurs with invasion to and subsequent proliferation of neoplastic cells in the subarachnoid space. Malignancies of diverse origins may spread to this space, which is bound by the leptomeninges.

Spread of hematologic cancers to this space and direct CSF seeding of intraparenchymal intraaxial CNS tumors are also well recognized.

The leptomeninges consist of the arachnoid and the pia mater; the space between the two contains the CSF. When tumor cells enter the CSF (either by direct extension, as in primary brain tumors, or by hematogenous dissemination, as in leukemia), they are transported throughout the nervous system by CSF flow, causing either multifocal or diffuse infiltration of the leptomeninges in a sheetlike fashion along the surface of the brain and spinal cord. This multifocal seeding of the leptomeninges by malignant cells is LC. LC is often called lymphomatous meningitis or leukemic meningitis if the primary is not a solid tumor.

Meningeal symptoms are the first manifestations in some patients including headaches (usually associated with nausea, vomiting, light-headedness), gait difficulties from weakness or ataxia, memory problems, incontinence, and sensory abnormalities. Pain and seizures are the most common presenting complaints. CNS symptoms of LC are generally divided into three anatomic groups (1) cerebral involvement including headache, lethargy, papilledema, behavioral changes, and gait disturbance; (2) cranial-nerve involvement including impaired vision, diplopia, hearing loss, and sensory deficits, including vertigo; and cranial-nerve palsies; and (3) spinal-root involvement including nuchal rigidity and neck and back pain, or invasion of the spinal roots.

The prognosis for patients with LC is generally poor because LC usually signifies the presence of metastases elsewhere, and the course of the systemic cancer is the major determinant of the patient's survival. The exception is leukemic or lymphomatous meningitis, which is often sensitive to both methotrexate and cytarabin and often can be eradicated completely from the CNS. Among patients with LC from solid tumors, the best response to chemotherapy and radiation occurs in those with LC from breast cancer, with 60% improving or stabilizing and a median survival of 7 months; 15% survive for a year, a survival rate rare in patients with LC with a primary tumor other than breast. Only 40% of LCs from small-cell lung carcinoma improve or stabilize, and patients with this disease have a median survival of only 4 months. Melanoma-derived LC has the worst prognosis with a 3.6-month median survival, and only 20% of these patients stabilize or improve with treatment. Nonresponders to chemotherapy seldom survive longer than a month. The prognosis for LC has not improved significantly in the last 20 years despite an increase in incidence and diagnosis.

Standard Therapy for LC

Leptomeningeal carcinomatosis is generally considered incurable and is difficult to treat. Treatment goals generally include improvement or stabilization of the patients neurologic status, prolongation of survival, and palliation. Most patients require a combination of surgery, radiation, and chemotherapy. Standard therapies include radiation therapy to symptomatic sites and regions where imaging has demonstrated bulk disease and intrathecal chemotherapy (e.g., methotrexate, cytarabin, thiotepa). Radiation palliates local symptoms, relieves CSF flow obstruction, and treats areas such as nerve-root sleeves, Virchow-Robin spaces, and the interior of bulky lesions that chemotherapy does not reach. Intrathecal chemotherapy treats subclinical leptomeningeal deposits and tumor cells floating in the CSF, preventing further seeding. Supportive care for patients includes analgesia with opioids, anticonvulsants for seizures, antidepressants, and anxiolytics. Attention problems and somnolence from whole-brain radiation may be treated with psychostimulants or modafinil.

Treatment of Drug Resistant or Refractory Cancer

The patient being treated in a method of the present invention may have a cancer and/or LC that is drug resistant or refractory. Because the conjugates of the invention have activity even in cancers that have demonstrated resistance to standard chemotherapeutic agents, the methods of the invention are particularly useful in treating such drug resistant cancers and/or LC.

Drug resistance typically arises following treatment with a particular chemotherapeutic. Multiple drug resistance (MDR) can arise when a cell overproduces the p-glycoprotein (P-gp) efflux transporter. As many chemotherapeutic drugs can be P-gp substrates, including vinblastine, doxorubicin, etoposide, colchicine, and paclitaxel, overexpression of P-gp in a cancer cell can lead to broad spectrum of resistance toward chemotherapeutic agents.

The present inventors have previously shown that paclitaxel conjugated to Angiopep-1 or Angiopep-2 are not P-gp substrates and thus should not be sensitive to P-gp overexpression in tumor cells; see, e.g., pages 46-47 and FIG. 9A of International Application Publication WO 2007/009229. Thus, the drug conjugates described herein are useful in treating patients having cancer and/or LC that is resistant to standard chemotherapeutic drugs.

Enhanced Uptake into LRP Expressing Cells

The methods of the invention may be especially useful in treating cancers having cells that express low density lipoprotein-related protein (LRP) receptor. The LRP receptor is expressed on the surface of cells, and is capable of binding to various substrates including aprotinin. The polypeptides described herein were designed based on the consensus kunitz-domain sequences that act as LRP receptor ligands (see, e.g., PCT Publication No. WO 2004/060403). Uptake of the conjugates including Angiopep-1 or Angiopep-2 is inhibited by LRP ligands, thus indicating involvement of LRP in this process. Specifically, the LRP ligands RAP (200 nM) and aprotinin (10 μM) are capable of reducing brain uptake of an Angiopep conjugate. Angiopep-2 (10 or 100 μM) is similarly able to reduce uptake of the conjugates into cells.

The blood-CSF barrier has been shown to express LRP (see Fujiyoshi et al. Journal of Neurochemistry, 2011, 118:407-415). Accordingly, LC is well suited for treatment using therapeutics that target LRP-expressing cells. As shown in FIG. 1, an Angiopep-2 conjugate is capable of accumulating in the meninges of a living mouse brain.

Combination Therapy

The methods of the invention may include administration of second therapeutic agent or treatment with a second therapy (e.g., a therapeutic agent or therapy that is standard in the art). Exemplary therapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anakinra, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacuzimab, bexarotene, bleomycin, bleomycin, bortezombi, bortezomib, busulfan, busulfan, calusterone, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, actinomycin D, dalteparin (e.g., sodium), darbepoetin alfa, dasatinib, daunorubicin, daunomycin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin (e.g., HCl), epoetin alfa, erlotinib, estramustine, etoposide (e.g., phosphate), exemestane, fentanyl (e.g., citrate), filgrastim, floxuridine, fludarabine, fluorouracil, 5-FU, fulvestrant, gefitinib, gemcitabine (e.g., HCl), gemtuzumab ozogamicin, goserelin (e.g., acetate), histrelin (e.g., acetate), hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib (e.g., mesylate), Interferon alfa-2b, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide (e.g., acetate), levamisole, lomustine, CCNU, meclorethamine (nitrogen mustard), megestrol, melphalan (L-PAM), mercaptopurine (6-MP), mesna, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate, panitumumab, pegademase, pegaspargase, pegfilgrastim, peginterferon alfa-2b, pemetrexed (e.g., disodium), pentostatin, pipobroman, plicamycin (mithramycin), porfimer (e.g., sodium), procarbazine, quinacrine, rasburicase, rituximab, sargramostim, sorafenib, streptozocin, sunitinib (e.g., maleate), talc, tamoxifen, temozolomide, teniposide (VM-26), testolactone, thalidomide, thioguanine (6-TG), thiotepa, thiotepa, thiotepa, topotecan (e.g., HCl), toremifene, Tositumomab/I-131 (tositumomab), trastuzumab, trastuzumab, tretinoin (ATRA), uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, zoledronate, and zoledronic acid. Exemplary derivatives of paclitaxel are described in U.S. Pat. No. 6,911,549, the entire contents of which are hereby incorporated by reference. Other agents include that can be used include antiestrogen agents such as tamoxifen (e.g., citrate), raloxifene, toremifene, and SCH 57068.

Polypeptide Conjugates

The methods of the invention include administration of a peptide-anticancer agent conjugate, such as those described in U.S. Patent Applications Publication Nos. 2006/0182684, and 2006/0189515, and U.S. Provisional Application No. 61/008,880, filed Dec. 20, 2007. Such conjugates may include any polypeptide described herein, an agent capable of treating LC such as paclitaxel or a paclitaxel analog (e.g., those described herein), and a linker (e.g., those described herein). Paclitaxel conjugates are exemplified by ANG1005, which includes the AngioPep-2 peptide (SEQ ID NO:97) conjugated to three paclitaxel molecules through ester linkages at the N-terminus, and through lysines at positions 10 and 15.

The conjugates, in certain embodiments, can cross the blood-brain barrier (BBB), the blood-CSF barrier, or can be preferentially targeted to certain cell types, such as breast, ovary, liver, lung, kidney, muscle cells or may be targeted to tumor cells (of any cell type described herein). The agents conjugated to these peptides can exhibit increased uptake across the BBB, which is mainly formed by endothelial cells with tight junctions, and blood-CSF barriers, which in contrast to the BBB is mainly formed by epithelial cells, and into the targeted cells, for example, by receptor-mediated endocytosis (e.g., through an LRP receptor). The conjugated agents may, either alternatively or in addition, exhibit increased stability or reduced expulsion from the cell (e.g., due to P-glycoprotein mediated efflux). Conjugates may further have activity in cancer cells that are resistant to standard chemotherapies.

Conjugates

The polypeptides described herein or derivatives thereof are conjugated to an anticancer agent (e.g., any known in the art). Each polypeptide may be conjugated to at least 1, 2, 3, 4, 5, 6, or 7 agents. In other embodiments, each agent has at least 1, 2, 3, 4, 5, 6, 7, 10, 15, 20, or more polypeptides attached thereto. The conjugates of the invention may be able to promote accumulation (e.g., due to increased uptake or reduced removal) of the agent in a particular cell type or tissue such as ovary, liver, lung, kidney, spleen or muscle of a subject.

The agent may be releasable from the vector after transport into a particular cell type or across the BBB. The agent can be released, for example, by enzymatic cleavage or other breakage of a chemical bond between the vector and the agent. The released agent may then function in its intended capacity in the absence of the vector.

In particular embodiments, the agent is paclitaxel or a paclitaxel analog (e.g., those described herein). Other anticancer agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anakinra, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG Live, bevacuzimab, bexarotene, bleomycin, bleomycin, bortezombi, bortezomib, busulfan, busulfan, calusterone, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, actinomycin D, dalteparin (e.g., sodium), darbepoetin alfa, dasatinib, daunorubicin, daunomycin, decitabine, denileukin, Denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin (e.g., HCl), epoetin alfa, erlotinib, estramustine, etoposide (e.g., phosphate), exemestane, fentanyl (e.g., citrate), filgrastim, floxuridine, fludarabine, fluorouracil, 5-FU, fulvestrant, gefitinib, gemcitabine (e.g., HCl), gemtuzumab ozogamicin, goserelin (e.g., acetate), histrelin (e.g., acetate), hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib (e.g., mesylate), Interferon alfa-2b, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide (e.g., acetate), levamisole, lomustine, CCNU, meclorethamine (nitrogen mustard), megestrol, melphalan (L-PAM), mercaptopurine (6-MP), mesna, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate, panitumumab, pegademase, pegaspargase, pegfilgrastim, peginterferon alfa-2b, pemetrexed (e.g., disodium), pentostatin, pipobroman, plicamycin (mithramycin), porfimer (e.g., sodium), procarbazine, quinacrine, rasburicase, rituximab, sargramostim, sorafenib, streptozocin, sunitinib (e.g., maleate), talc, tamoxifen, temozolomide, teniposide (VM-26), testolactone, thalidomide, thioguanine (6-TG), thiotepa, thiotepa, thiotepa, topotecan (e.g., hcl), toremifene, Tositumomab/I-131 (tositumomab), trastuzumab, trastuzumab, tretinoin (ATRA), uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, zoledronate, and zoledronic acid.

Other anticancer agents include antibodies. Conjugation of such antibodies may be accomplished using any means known in the art (e.g., using the conjugation strategies described herein). Any diagnostic or therapeutic antibody may be conjugated to one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) vectors of the invention. In addition, antibody fragments (e.g., capable of binding to an antigen) may also be conjugated to the vectors of the invention. Antibody fragments include the Fab and Fc regions, heavy chain, and light chain of an antibody (e.g., of any antibody described herein). Exemplary antibodies for use in diagnosis and therapy of cancer include ABX-EGF (Panitimumab), OvaRex (Oregovemab), Theragyn (pemtumomabytrrium-90), Therex, Bivatuzumab, Panorex (Edrecolomab), ReoPro (Abciximab), Bexxar (Tositumomab), MAb, idiotypic 105AD7, Anti-EpCAM (Catumaxomab), MAb lung cancer (from Cytoclonal), Herceptin (Trastuzumab), Rituxan (Rituximab), Avastin (Bevacizumab), AMD Fab (Ranibizumab), E-26 (2^nd gen. IgE) (Omalizumab), Zevalin (Rituxan+yttrium-90) (Ibritumomab tiuxetan), Cetuximab, BEC2 (Mitumomab), IMC-1C11, nuC242-DM1, LymphoCide (Epratuzumab), LymphoCide Y-90, CEA-Cide (Labetuzumab), CEA-Cide Y-90, CEA-Scan (Tc-99m-labeled arcitumomab), LeukoScan (Tc-99m-labeled sulesomab), LymphoScan (Tc-99m-labeled bectumomab), AFP-Scan (Tc-99m-labeled), HumaRAD-HN (+yttrium-90), HumaSPECT (Votumumab), MDX-101 (CTLA-4), MDX-210 (her-2 overexpression), MDX-210/MAK, Vitaxin, MAb 425, IS-IL-2, Campath (alemtuzumab), CD20 streptavidin, Avidicin, (albumin+NRLU13), Oncolym (+iodine-131) Cotara (+iodine-131), C215 (+staphylococcal enterotoxin, MAb lung/kidney cancer (from Pharmacia Corp.), nacolomab tafenatox (C242 staphylococcal enterotoxin), Nuvion (Visilizumab), SMART M195, SMART 1D10, CEAVac, TriGem, TriAb, NovoMAb-G2 radiolabeled, Monopharm C, GlioMAb-H (+gelonin toxin), Rituxan (Rituximab), and ING-1. Additional therapeutic antibodies include 5G1.1 (Ecluizumab), 5G1.1-SC (Pexelizumab), ABX-CBL (Gavilimomab), ABX-IL8, Antegren (Natalizumab), Anti-CD11a (Efalizumab), Anti-CD18 (from Genetech), Anti-LFA1, Antova, BTI-322, CDP571, CDP850, Corsevin M, D2E7 (Adalimumab), Humira (Adalimumab), Hu23F2G (Rovelizumab), ICI4, IDEC-114, IDEC-131, IDEC-151, IDEC-152, Infliximab (Remicade), LDP-01, LDP-02, MAK-195F (Afelimomab), MDX-33, MDX-CD4, MEDI-507 (Siplizumab), OKT4A, OKT3 (Muromonab-CD3), and ReoPro (Abciximab).

Conjugation Linkers

The conjugate used in the invention may include using any cross-linking (conjugation) reagent or protocol known in the art, many of which are commercially available. Such protocols and reagents include, cross-linkers reactive with amino, carboxyl, sulfhydryl, carbonyl, carbohydrate and/or phenol groups. The amounts, times, and conditions of such protocols can be varied to optimize conjugation. Cross-linking reagents contain at least two reactive groups and are generally divided into homofunctional cross-linkers (containing identical reactive groups) and heterofunctional cross-linkers (containing non-identical reactive groups). The cross-linkers of the invention may be either homobifunctional and/or heterobifunctional. Furthermore the cross-linker may incorporate a ‘spacer’ between the reactive moieties, or the two reactive moieties in the cross-linker may be directly linked. Bonds may include ester bonds.

Exemplary linkers include BS³ [Bis(sulfosuccinimidyl)suberate], NHS/EDC (N-hydroxysuccinimide and N-ethyl-(dimethylaminopropyl)carbodimide, Sulfo-EMCS ([N-e-Maleimidocaproic acid]hydrazide), SATA (N-succinimidyl-S-acetylthioacetate), and hydrazide. BS³ is a homobifunctional N-hydroxysuccinimide ester that targets accessible primary amines. NHS/EDC allows for the conjugation of primary amine groups with carboxyl groups. Sulfo-EMCS are heterobifunctional reactive groups (maleimide and NHS-ester) that are reactive toward sulfhydryl and amino groups. Amine coupling using sulfo-NHS/EDC activation may be used to cross-link therapeutic antibodies to polypeptides. The resulting conjugate may be stable and retain the biological activity of the antibody. Moreover, it has a high conjugation capacity that can be reliably controlled and a low non-specific interaction during coupling procedures. SATA is reactive towards amines and adds protected sulfhydryls groups. The NHS-ester reacts with primary amines to form stable amide bonds. Sulfhydryl groups may be deprotected using hydroxylamine. Hydrazide can be used to link carboxyl groups to primary amines and may therefore be useful for linking glycoproteins.

Small molecules such as therapeutic agents can be conjugated to polypeptides (e.g., those described herein). The exemplary small molecule, paclitaxel, has two strategic positions (position C2′ and C7) useful for conjugation. Conjugation of a vector or vector of the invention to paclitaxel can be performed as follows. Briefly, paclitaxel is reacted with anhydride succinic pyridine for three hours at room temperature to attach a succinyl group in position 2′. The 2′-succinyl paclitaxel has a cleavable ester bond in position 2′ can simply release succinic acid. This cleavable ester bond can be further used for various modifications with linkers, if desired. The resulting 2′-O-succinyl-paclitaxel is then reacted with EDC/NHS in DMSO for nine hours at room temperature, followed by the addition of the vector or vector in Ringer/DMSO for an additional reaction time of four hours at room temperature. Each intermediate, such as paclitaxel, 2′-O-succinyl-paclitaxel and 2′-O-NHS-succinyl-paclitaxel, is purified and validated using different approaches such as HPLC, thin liquid chromatography, NMR (¹³C or ¹H exchange), melting point, mass spectrometry. The final conjugate is analyzed by mass spectrometry and SDS-polyacrylamide gel electrophoresis. This allows determining the number of paclitaxel molecules conjugated on each vector.

Dosages

The dosage of any conjugate or composition described herein depends on several factors, including: the administration method, the severity of the disease, whether the cancer is to be treated or prevented, and the age, weight, and health of the subject to be treated.

With respect to the treatment methods of the invention, it is not intended that the administration of a vector, conjugate, or composition to a subject be limited to a particular mode of administration, dosage, or frequency of dosing; the invention contemplates all modes of administration. The conjugate, or composition may be administered to the subject in a single dose or in multiple doses. For example, a compound described herein or identified using screening methods of the invention may conjugate be administered once a week for, e.g., 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, or more weeks. It is to be understood that, for any particular subject, specific dosage regimes should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the composition. For example, the dosage of a composition can be increased if the lower dose does not provide sufficient activity in the treatment of a disease or condition described herein (e.g., cancer and/or LC). Conversely, the dosage of the composition can be decreased if the disease (e.g., cancer and/or LC) is reduced or eliminated.

While the attending physician ultimately will decide the appropriate amount and dosage regimen, a therapeutically effective amount of a vector, conjugate, or composition described herein, may be, for example, in the range of 0.0035 μg to 20 μg/kg body weight/day or 0.010 μg to 140 μg/kg body weight/week. Desirably a therapeutically effective amount is in the range of 0.025 μg to 10 μg/kg, for example, at least 0.025, 0.035, 0.05, 0.075, 0.1, 0.25, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, or 9.0 μg/kg body weight administered daily, every other day, or twice a week. In addition, a therapeutically effective amount may be in the range of 0.05 μg to 20 μg/kg, for example, at least 0.05, 0.7, 0.15, 0.2, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 10.0, 12.0, 14.0, 16.0, or 18.0 μg/kg body weight administered weekly, every other week, every three weeks or once a month. Furthermore, a therapeutically effective amount of a compound may be, for example, in the range of 0.1 mg/m² to 2,000 mg/m² administered every other day, once weekly, every other week or every three weeks. For example ANG1005, may be administered at 50, 100, 200, 300, 400, 420, 500, 600, 650, 700, 800, or 1,000 mg/m² every one, two, three, four weeks, or every month or every other month. In one particular example, ANG1005 is administered at between 300 mg/m² and 650 mg/m² every three weeks. In another embodiment, the therapeutically effective amount is in the range of 1000 μg/m² to 20,000 μg/m², for example, at least 1000, 1500, 4000, or 14,000 μg/m² of the compound administered daily, every other day, twice weekly, weekly, or every other week.

Formulation of Pharmaceutical Compositions

The administration of a conjugate described herein or a composition containing the conjugate may be by any suitable means that results in a concentration of the compound that treats LC. The conjugate may be in any appropriate amount of any suitable carrier substance, and is generally present in an amount of 1-95% by weight of the total weight of the composition. The composition may be provided in a dosage form that is suitable for the oral, parenteral (e.g., intravenously or intramuscularly), rectal, cutaneous, nasal, vaginal, inhalant, skin (patch), topical, ocular, or intracranial administration route. Thus, the composition may be in the form of, e.g., tablets, capsules, pills, powders, granulates, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, drenches, osmotic delivery devices, suppositories, enemas, injectables, implants, sprays, or aerosols. The pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see, e.g., Remington: The Science and Practice of Pharmacy, 20th edition, 2000, ed. A. R. Gennaro, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York).

Pharmaceutical compositions may be formulated to release the conjugate(s) immediately upon administration or at any predetermined time or time period after administration. The latter types of compositions are generally known as controlled release formulations, which include (i) formulations that create substantially constant concentrations of the conjugate(s) within the body over an extended period of time; (ii) formulations that after a predetermined lag time create substantially constant concentrations of the conjugate(s) within the body over an extended period of time; (iii) formulations that sustain the conjugate(s) action during a predetermined time period by maintaining a relatively constant, effective level of the conjugate(s) in the body with concomitant minimization of undesirable side effects associated with fluctuations in the plasma level of the conjugate(s) (sawtooth kinetic pattern); (iv) formulations that localize action of conjugate(s), e.g., spatial placement of a controlled release composition adjacent to or in the diseased tissue or organ; (v) formulations that achieve convenience of dosing, e.g., administering the composition once per week or once every two weeks; and (vi) formulations that target the action of the conjugate(s) by using carriers or chemical derivatives to deliver the compound to a particular target cell type. Administration of the conjugate(s) in the form of a controlled release formulation is especially preferred for conjugate(s) having a narrow absorption window in the gastro-intestinal tract or a relatively short biological half-life.

Any of a number of strategies can be pursued in order to obtain controlled release in which the rate of release outweighs the rate of metabolism of the conjugate(s) in question. In one example, controlled release is obtained by appropriate selection of various formulation parameters and ingredients, including, e.g., various types of controlled release compositions and coatings. Thus, the conjugate(s) is formulated with appropriate excipients into a pharmaceutical composition that, upon administration, releases the conjugate(s) in a controlled manner. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, molecular complexes, microspheres, nanoparticles, patches, and liposomes.

EXAMPLES

Example 1. Treatment of LC in Breast Cancer Subjects

Method:

Twenty-eight subjects diagnosed with LC were administered 600 mg/m² of ANG1005 intravenously on a tri-weekly schedule. The subjects received between one and nine cycles of ANG1005. The receptor status of the subjects is shown in Table

TABLE 1
Receptor Status of Subjects
            Positive Patients/
Marker      Negative Patients
HER2+ 17/28 17/28
HER/− 11/28 11/28
ER+ 16/28   16/28
ER− 12/28   12/28
PgR+ 13/28  13/28
PgR− 15/28  15/28

Results:

The results of the treatment of the subjects according to CNS Response Evaluation Criteria in Solid Tumors (RECIST) based on brain parenchymal tumor response is shown in Table 2.

TABLE 2
Subject Outcomes
	All
	Patients	HER2+	HER2−
Outcome	(n = 23)	(n = 15)	(n = 8)
Partial Response (PR)	5	(22%)	4	(27%)	1	(12.5%)
Stable Disease (SD)	12	(52%)	8	(53%)	4	(50%)
Progressive Disease	6	(26%)	3	(20%)	3	(37.5%)
Patient Benefit (PR + SD)	17	(74%)	12	(80%)	5	(62.5%)

As shown in Table 2, the patient benefit response rate, which includes subjects displaying a partial response or stable disease was 74%. The patient benefit response rate was 80% for subjects with HER2 positive breast cancer.

Example 2. Treatment of Subject 2

Subject 2 is a 59 year old female diagnosed in October 2012 with HER2+/ER−/PgR+infiltrating ductal carcinoma. In January 2014, Subject 2 was diagnosed with brain metastases, and in October 2014 the brain metastases relapsed with leptocarcinomatosis. Subject 2 previously received several courses of therapy including cytoxan, taxotere, and herceptin from November 2012 to March 2013, herceptin from March 2013 to October 2013, a right mastectomy in May 2013, craniotomy and SRS in February 2014, craniotomy in July 2014, and nevatinib and capecitabine in August 2014.

Subject 2 presented at the time of treatment with ANG1005 with an active tumor extending from the deep portion of the surgical cavity to the right tentorial surface and along the lateral right temporal occipital dural surface. Subject 2 started treatment with ANG1005 in October 2014 and received 7 cycles (once every three weeks). Treatment was terminated due to clinical disease progression.

The leptomeningeal carcinomatosis of Subject 2 responded to ANG1005 as indicated by a lesion in the lateral right middle cranial fossa dural surface being no longer apparent post-treatment, and a nodule of leptomeningeal metastasis just superior to the right tentorial leaflet in the inferior surface of the right hippocampal gyrus has decreasing in size.

Example 3. Treatment of Subject 3

Subject 3 is a 44 year old female diagnosed in May 2009 with HER2+/ER−/PgR− infiltrating ductal carcinoma. In March 2012, Subject 3 was diagnosed with brain metastases, and in September 2014 suffered a recurrence of the brain metastases. Subject 3 has previously received several courses of therapy including abraxane and lapatinib from May to August 2009, vinoblastine and trastuzumab from August 2012 to April 2013, WRBT in September 2012, TDM1 from April to July 2013, capecitabine and lapatinib from April to July 2014, and capecitabine and TDM1 in September 2014.

Subject 3 presented at the time of treatment with ANG1005 with numerous (>10) brain metastases all of which had increased in size despite prior treatment. Extensive scattered osseous metastatic disease in head/neck. After presenting with a numb chin (left trigeminal cranial nerve deficits), Subject 3 was also found to have leptomeningeal disease. Subject 3 started treatment with ANG1005 in September 2014 and received 5 cycles (once every three weeks). Treatment was terminated due to adverse event (pneumonia).

The leptomeningeal carcinomatosis of Subject 3 improved both radiographically and clinically. Subject 3 was able to ambulate better and had no cranial nerve or bowel/bladder symptoms after the first two cycles. As shown in FIGS. 2A and B, both the brain metastases and LC of Subject 3 responded to treatment with ANG1005.

Example 4. Treatment of Subject 4

Subject 4 is a 58 year old female diagnosed in October 2011 with HER2+/ER−/PgR− ductal carcinoma in situ. In November 2012, Subject 4 was diagnosed with brain metastases, and in March 2015 the brain metastases relapsed with leptomeningeal carcinomatosis. Subject 4 previously received several courses of therapy including carboplatin from October to November 2011, denosumab from November 2011 to November 2012, paclitaxel from December 2011 to March 2012, WBRT, transtuzumab from May 2012 to November 2012, and kadcyla from April 2013 to December 2014. Subject 4 presented at the time of treatment with ANG1005 with parenchymal metastases present in the cerebellum and widespread LC of the cerebellum and cerebral hemispheres. In addition, Subject 4 presented with mild ventricular enlargement with compression upon the fourth ventricle and cutflow tracts. Subject 4 started treatment with ANG1005 in March 2015 and received 8 cycles (once every three weeks). Subject withdrew from treatment due to low performance scores.

The leptomeningeal carcinomatosis of Subject 4 responded to ANG1005 as indicated by diminished size and extent of nodular leptomeningeal enhancement related to the cerebellum with result effacement of the fourth ventricle. Furthermore, Subject 4 displayed interval decrease in volume of leptomeningeal tumor burden post-treatment.

Example 5. Treatment of Subject 8

Subject 8 is a 42 year old female diagnosed in April 2014 with HER2+/ER+/PgR− infiltrating ductal carcinoma. In December 2014, Subject 8 was diagnosed with brain metastases and in March 2015 the brain metastases recurred with leptomeningeal carcinomatosis. Subject 8 previously received several courses of therapy including docetaxel from May to October 2014, trastuzumab and pertuzumab from May to December 2014, and capecitabine and lapatinib from January to April 2015.

Subject 8 presented at the time of treatment with ANG1005 with numerous enhancing lesions throughout the supratentorial and infratentorial brain, presence of supratentorial white matter lesions, suggestive of demyelinating disease of unknown origin, CSF negative for malignant cells, and multiple stable metastases in liver. Subject 8 started treatment with ANG1005 in April 2015 and received 2 cycles (once every three weeks). Subject 8 terminated from treatment in July 2015 due to clinical progression.

The leptomeningeal carcinomatosis of Subject 8 responded to ANG1005 as indicated by improvement in extensive LC in the cerebellum.

Example 6. Kaplan-Meier Estimates of Survival for Subjects with LC

The Kaplan-Meier method was used to estimate the overall survival of LC subjects treated with ANG1005. As shown in FIG. 4, treatment with ANG1005 was estimated to increase median overall survival compared to the historical median overall survival of 3-6 months, following aggressive treatment.

Example 7. Treatment Protocol

All patients will receive ANG1005 at a starting dose of 600 mg/m² by intravenous (IV) infusion once every 3 weeks (1 cycle). Dose reductions or delays will be allowed at any dosing cycle if toxicity is observed. Patients will be monitored during infusion and for a minimum of 1 hour after the completion of each infusion.

Intracranial and extracranial tumor evaluation by MRI/CT will be performed at baseline and after every 2 cycles (i.e., every 6±2 weeks). If a partial or complete response is seen, the subsequent MRI should be conducted at 4 weeks but 6 weeks for confirmation of response. Patients who develop extracranial disease progression in the absence of intracranial disease progression, should be removed from the study, unless the following conditions are met: (1) There is evidence of clinical benefit attributed to ANG1005 therapy, such as: (a) clinical improvement in symptoms from brain metastases, (b) radiographic improvement of brain metastases, and (2) the systemic progression is asymptomatic. However, if the Investigator decides to initiate a non-protocol systemic anti-cancer therapy as needed, then ANG1005 will be discontinued. Patients whose study treatment is discontinued before the 1 year maximum treatment period and whose disease has not progressed will continue to be followed for disease progression. Disease assessments including radiographic assessments will be done at approximately 8-week intervals, or per the institutional standard practice, starting from the date of the last dose of study treatment until intracranial and extracranial disease progression is documented. Survival follow-up will be done at approximately 8-week intervals from the date of the last dose. Patients will remain on study treatment under this protocol for a maximum period of one year unless they develop disease progression or develop unacceptable toxicity. Further treatment beyond the one year maximum period will be considered on a case-by-case basis.

OTHER EMBODIMENTS

While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth.

Claims

1. A method for the treatment of leptomeningeal carcinomatosis comprising administering to a subject in need thereof an effective amount of a compound having the structure:

2. The method of claim 1, wherein the primary origin of said leptomeningeal carcinomatosis is a solid tumor.

3. The method of claim 2, wherein said solid tumor is a breast tumor, a lung tumor, a gastrointestinal tumor, or malignant melanoma.

4. The method of claim 3, wherein said solid tumor is a breast tumor.

5. The method of claim 4, wherein said breast tumor has been identified as a HER2 positive tumor.

6. The method of claim 4, wherein said breast tumor has been identified as a triple negative tumor.

7. The method of claim 2, wherein said tumor comprises cells that express MDR1.

8. The method of claim 2, wherein said subject previously received another anticancer therapy for the leptomeningeal carcinomatosis and/or the solid tumor.

9. The method of claim 8, wherein said anticancer therapy comprises a chemotherapeutic agent.

10. The method of claim 9, wherein said chemotherapeutic agent is a taxane, a platinum-based agent, an anthracycline, an anthraquinone, an alkylating agent, a HER2 targeting therapy, vinorelbine, a nucleoside analog, ixabepilone, eribulin, cytarabine, a hormonal therapy, capecitabine, lapatinib, 5-FU, vincristine, etoposide, or methotrexate.

11. The method of claim 8, wherein the solid tumor and/or leptomeningeal carcinomatosis failed to respond to said previously received anticancer therapy and/or relapsed after treatment with said previously received anticancer therapy.

12. The method of claim 1, wherein said method further comprises administration of an additional anticancer therapy.

13. The method of claim 12, wherein said additional anticancer therapy comprises radiation therapy and/or a chemotherapeutic agent.

14. The method of claim 13, wherein said additional anticancer therapy comprises radiation therapy.

15. The method of claim 13, wherein said chemotherapeutic agent is a taxane, a platinum-based agent, an anthracycline, an anthraquinone, an alkylating agent, a HER2 targeting therapy, vinorelbine, a nucleoside analog, ixabepilone, eribulin, cytarabine, a hormonal therapy, or methotrexate.

16. The method of claim 15, wherein said chemotherapeutic agent is methotrexate, an alkylating agent, cytarabine, or a HER2 targeting therapy.

17. The method of claim 1, wherein said method further comprises administration of a palliative therapy.

18. The method of claim 17, wherein said palliative therapy is an analgesic, an anticonvulsant, an antidepressant, an anxiolytic, a psychostimulant, modafinil, palliative radiation, corticosteroids, an H1 antagonist, a hematopoietic growth factor, and/or a blood transfusion.

19. The method of claim 4, wherein said breast tumor is a HER2 negative tumor.