PH-responsive astemizole nanoparticles and methods for making and using the same

Abstract

In accordance with the purpose(s) of the disclosure, as embodied and broadly described herein, the disclosure, in an aspect, relates to pH-responsive nanoparticles described herein facilitate the release of astemizole to effectively release astemizole locally, preventing or reducing cancer cell growth, while minimizing effects on healthy cells. The compositions comprise a pH-responsive nanoparticle having a core comprising astemizole or a pharmaceutically acceptable salt thereof encapsulated by a pH-responsive polymer carrier. The pH-responsive nanoparticles can be administred alone or in combination with other other pharmaceutical compounds such as, for example, anticancer agent.

Description

ATTORNEY DOCKET NO.930307-2000 PH-RESPONSIVE ASTEMIZOLE NANOPARTICLES AND METHODS FOR MAKING AND USING THE SAME CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to co-pending U.S. Provisional Patent Application No.63 / 581,016, filed on September 7, 2023, the contents of which are incorporated by reference herein in their entireties. BACKGROUND

[0002] Currently, chemotherapy and radiation therapy remain the most common methods for cancer treatment. These treatments were based on targeting proliferating cells rather than cancer cells only, which is also a factor in detrimental side effects from these treatments. Targeted therapy, a new generation of cancer treatment, targets cancer-specific changes of molecules and signaling pathways to inhibit cellular proliferation or induce cancer cell death, but with limited side effects on normal cells. Considerable effort has been made in finding the targets and the ways of affecting the targets inside the cells as a treatment, in particular for targets associated with difficult to treat cancers such as pancreatic cancer. For example, pancreatic ductal adenocarcinoma (PDAC) remains one of the most deadly cancers with an average survival of 3.5 months for non- resectable tumors. Gemcitabine is the primary prescribed chemotherapy drug for PDAC, but with only limited success.

[0003] Angiopoietin Like 4 (ANGPTL4) expression has been implicated in many cancer types and other diseases both as pathogenic and beneficial. It is known to occur in at least three isoforms, each with different functions. Previous work (Kirby, M.K., et al., 2016, Molecular Oncology, 10(8): 1169–1182), has shown varying gene expression levels for PDAC tumors correlating with survival outcomes. This study assessed RNA sequencing data from 119 patient tumor tissues, and 323 survival-correlated transcripts were identified. Further analysis revealed a 19-transcript predictive model for survival rates. In vitro patient tumor cell studies revealed a subset of these transcripts, which correlated with resistance to gemcitabine.

[0004] Despite advances in research directed towards improving outcomes with cancer, there is still a scarcity of compounds that are both potent, efficacious, and selective modulators of targets in cancer cells, such as ANGPTL4. These needs and other needs are satisfied by theATTORNEY DOCKET NO.930307-2000 present disclosure. SUMMARY

[0005] In accordance with the purpose(s) of the disclosure, as embodied and broadly described herein, the disclosure, in an aspect, relates to pH-responsive nanoparticles described herein facilitate the release of astemizole to effectively release astemizole locally, preventing or reducing cancer cell growth, while minimizing effects on healthy cells. The compositions comprise a pH-responsive nanoparticle having a core comprising astemizole or a pharmaceutically acceptable salt thereof encapsulated by a pH-responsive polymer carrier. The pH-responsive nanoparticles can be administred alone or in combination with other other pharmaceutical compounds such as, for example, anticancer agents.

[0006] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class. Unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification. DESCRIPTION

[0007] Many modifications and other embodiments disclosed herein will come to mind to one skilled in the art to which the disclosed compositions and methods pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the disclosures are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.ATTORNEY DOCKET NO.930307-2000

[0008] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0009] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure.

[0010] Any recited method can be carried out in the order of events recited or in any other order that is logically possible. That is, unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0011] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided herein can be different from the actual publication dates, which can require independent confirmation.

[0012] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class.

[0013] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed compositions and methods belong. It will be further understood that terms, such as those defined in commonly used dictionaries, should beATTORNEY DOCKET NO.930307-2000 interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0014] Prior to describing the various aspects of the present disclosure, the following definitions are provided and should be used unless otherwise indicated. Additional terms may be defined elsewhere in the present disclosure. Definitions

[0015] As used herein, “comprising” is to be interpreted as specifying the presence of the stated features, integers, steps, or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps, or components, or groups thereof. Moreover, each of the terms “by”, “comprising,” “comprises”, “comprised of,” “including,” “includes,” “included,” “involving,” “involves,” “involved,” and “such as” are used in their open, non-limiting sense and may be used interchangeably. Further, the term “comprising” is intended to include examples and aspects encompassed by the terms “consisting essentially of” and “consisting of.” Similarly, the term “consisting essentially of” is intended to include examples encompassed by the term “consisting of.

[0016] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an excipient” include, but are not limited to, mixtures or combinations of two or more such excipients, and the like.

[0017] It should be noted that ratios, concentrations, amounts, and other numerical data can be expressed herein in a range format. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms a further aspect. For example, if the value “about 10” is disclosed, then “10” is also disclosed.ATTORNEY DOCKET NO.930307-2000

[0018] When a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. For example, where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure, e.g. the phrase “x to y” includes the range from ‘x’ to ‘y’ as well as the range greater than ‘x’ and less than ‘y’. The range can also be expressed as an upper limit, e.g. ‘about x, y, z, or less’ and should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘less than x’, less than y’, and ‘less than z’. Likewise, the phrase ‘about x, y, z, or greater’ should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘greater than x’, greater than y’, and ‘greater than z’. In addition, the phrase “about ‘x’ to ‘y’”, where ‘x’ and ‘y’ are numerical values, includes “about ‘x’ to about ‘y’”.

[0019] It is to be understood that such a range format is used for convenience and brevity, and thus, should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. To illustrate, a numerical range of “about 0.1% to 5%” should be interpreted to include not only the explicitly recited values of about 0.1% to about 5%, but also include individual values (e.g., about 1%, about 2%, about 3%, and about 4%) and the sub-ranges (e.g., about 0.5% to about 1.1%; about 5% to about 2.4%; about 0.5% to about 3.2%, and about 0.5% to about 4.4%, and other possible sub-ranges) within the indicated range. Thus, for example, if a component is in an amount of about 1%, 2%, 3%, 4%, or 5%, where any value can be a lower and upper endpoint of a range, then any range is contemplated between 1% and 5% (e.g., 1% to 3%, 2% to 4%, etc.).

[0020] As used herein, the terms “about,” “approximate,” “at or about,” and “substantially” mean that the amount or value in question can be the exact value or a value that provides equivalent results or effects as recited in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art such that equivalent results or effects are obtained. In some circumstances, the value that provides equivalent results or effects cannot be reasonably determined. In such cases, it is generally understood, as used herein, that “about” and “at or about” mean the nominal value indicated ±10% variation unless otherwise indicated or inferred.ATTORNEY DOCKET NO.930307-2000 In general, an amount, size, formulation, parameter or other quantity or characteristic is “about,” “approximate,” or “at or about” whether or not expressly stated to be such. It is understood that where “about,” “approximate,” or “at or about” is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise.

[0021] As used herein, nomenclature for compounds, including organic compounds, can be given using common names, IUPAC, IUBMB, or CAS recommendations for nomenclature. When one or more stereochemical features are present, Cahn-Ingold-Prelog rules for stereochemistry can be employed to designate stereochemical priority, E / Z specification, and the like. One of skill in the art can readily ascertain the structure of a compound if given a name, either by systemic reduction of the compound structure using naming conventions, or by commercially available software, such as ChemDraw®(PerkinElmer, Inc., Waltham, Massachussetts).

[0022] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a functional group,” “an alkyl,” or “a residue” includes mixtures of two or more such functional groups, alkyls, or residues, and the like.

[0023] A weight percent (wt. %) of a component, unless specifically stated to the contrary, is based on the total weight of the formulation or composition in which the component is included.

[0024] As used herein, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0025] Unless otherwise expressly stated, it is in no way intended that any method set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not actually recite an order to be followed by its steps or it is not otherwise specifically stated in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non- express basis for interpretation, including: matters of logic with respect to arrangement of steps or operational flow; plain meaning derived from grammatical organization or punctuation; and the number or type of embodiments described in the specification.

[0026] As used herein, “ANGPTL4,” “Angiopoietin-related protein 4,” or “Angiopoietin Like 4” refer to a protein encoded by a human gene located at chromosomal location 19p13.2. The termsATTORNEY DOCKET NO.930307-2000 are also inclusive of protein homologues of the human protein. The human gene encodes three isoforms which are generated by alternative mRNA splice variants. The three protein isoforms are designated in the UniProtKB / Swiss-Prot database with the identifiers: Q9BY76-1, Q9BY76-2, and Q9BY76-3. The canonical protein sequence is associated with UniProtKB / Swiss-Prot database with the identifiers, Q9BY76-1, which is a 406 amino acid protein with a molecular weight of 45,214 kD. The ANGPTL4 protein is a glycosylated, secreted protein containing a C- terminal fibrinogen domain. The secreted protein circulates as a homooligomer.

[0027] The homooligomer undergoes proteolytic processing to release its carboxyl fibrinogen- like domain, which circulates as a monomer. The C-terminal domain may be proteolytically- cleaved from the full-length secreted protein in vivo. The C-terminal domain is referred to as cANGPTL4. The natural cleavage site in vivo can occur between amino acids 162-229 of the canonical sequence.

[0028] As used herein, the term “cANGPTL4” refers to a C-terminal fragment of the canonical ANGPTL4 human protein sequence beginning at any amino acid position between 162-229, or homologues thereof. In an aspect, the terminal fragment, cANGPTL4, comprises amino acid residures 179-406 of the canonical ANGPTL4 human protein sequence, or homologues thereof. In an aspect, the terminal fragment, cANGPTL4, comprises amino acid residures 184-406 of the canonical ANGPTL4 human protein sequence, or homologues thereof. Unless specified otherwise, the use of “ANGPTL4” is inclusive of reference to both ANGPTL4 and cANGPTL4.

[0029] As used herein, the term “ANGPTL4 modulator” or “modulator of ANGPTL4” refers to a compound or agent that directly or indirectly modulates the activity of ANGPTL4, as an isolated protein, in a cell or in an animal, in particular a mammal, for example a human. For example, an ANGPTL4 modulator can alter the basal activity level of isolated ANGPTL4 in vitro or the activity of ANGPTL4 in a cell, in vivo or in vitro. In an aspect, an ANGPTL4 modulator can inhibit the activity of ANGPTL4 in a cell. The cell can be a mammalian cell line transfected with human ANGPTL4. The cell can be a mammalian cell line transfected with a rodent, e.g., rat, ANGPTL4. The cell can be a mammalian cell line transfected with a mammalian ANGPTL4. The cell can be a cancer cell-line such as PANC-1, ASPC 1, BXPC-3, or MIA-PaCa-2 cells.

[0030] As used herein, the term “modulation of ANGPTL4 activity” refers to modulation of either ANGPTL4 or cANGPTL4 activity or both, unless otherwise specified. Modulation of activity, as understood herein, is altering the activity of ANGPTL4 from a baseline level of activity, e.g.,ATTORNEY DOCKET NO.930307-2000 inhibition of a baseline or basal activity level. Accordingly, in some aspects, modulation can be inhibition of ANGPTL4 activity. The modulation, for example, inhibition, of ANGPTL4 activity in a cell can result in cell death or inhibition of cellular proliferation.

[0031] As used herein, the term “subject” can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term “subject” also includes domesticated animals (e.g., cats, dogs, etc.), livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mouse, rabbit, rat, guinea pig, fruit fly, etc.). The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be covered. In an aspect, the subject is a mammal. A patient refers to a subject afflicted with a disease or disorder. The term “patient” includes human and veterinary subjects. In an aspect of a disclosed method, the subject has been diagnosed with a need for treatment of one or more disorders of uncontrolled cellular proliferation associated with an ANGPTL4 dysfunction, e.g., aberrant expression of ANGPTL4. In an aspect of a disclosed method, the subject has been diagnosed with a need for modulation of ANGPTL4 activity. In an aspect of a disclosed method, the subject has been diagnosed with a disorder of uncontrolled cellular proliferation, e.g. cancer. In an aspect of a disclosed method, the subject has been identified with a disorder treatable by modulation of ANGPTL4 activity. In an aspect, a subject can be treated prophylactically with a compound or composition disclosed herein. In an aspect, the subject is a mammal such as a primate. In an aspect, the subject is a human.

[0032] As used herein, the term “treatment” refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. Treatment includes active treatment, that is, treatment directed specifically toward the improvement of the health of a subject with a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, treatment includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. In various aspects, treatment includes any treatment of aATTORNEY DOCKET NO.930307-2000 subject, including a mammal (e.g., a human), and includes: (i) preventing the disease from occurring in a subject that can be predisposed to the disease but has not yet been diagnosed as having it; (ii) inhibiting the disease, i.e., arresting its development; or (iii) relieving the disease, i.e., causing regression of the disease.

[0033] As used herein, the term “diagnosed” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by the compounds, compositions, or methods disclosed herein. For example, “diagnosed with a disorder treatable by modulation of ANGPTL4 activity” means having been subjected to a physical examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by a compound or composition that can modulate ANGPTL4 activity.

[0034] As used herein, the phrase “identified to be in need of treatment for a disorder,” or the like, refers to selection of a subject based upon need for treatment of the disorder. For example, a subject can be identified as having a need for treatment of a disorder (e.g., a disorder related to ANGPTL4 activity) based upon an earlier diagnosis by a person of skill and thereafter subjected to treatment for the disorder. It is contemplated that the identification can, in an aspect, be performed by a person different from the person making the diagnosis. It is also contemplated, in an aspect, that the administration can be performed by one who subsequently performed the administration.

[0035] As used herein, the terms “administering” and “administration” refer to any method or route of administration of a pharmaceutical preparation to a subject. Such methods are well known to those skilled in the art and include, but are not limited to, oral administration, transdermal administration, administration by inhalation, nasal administration, topical administration, intravaginal administration, ophthalmic administration, intraaural administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injectable such as intravenous administration, intra-arterial administration, intramuscular administration, and subcutaneous administration. Administration can be continuous or intermittent. In various aspects, a preparation can be administered therapeutically; that is, administered to treat an existing disease or condition. In further various aspects, a preparation can be administered prophylactically; that is, administered for prevention of a disease or condition.ATTORNEY DOCKET NO.930307-2000

[0036] The term “contacting” as used herein refers to bringing a disclosed compound and a cell, a target (e.g. ANGPTL4), or other biological entity together in such a manner that the compound can affect the activity of the target, either directly; i.e., by interacting with the target itself, or indirectly; i.e., by interacting with another molecule, co-factor, factor, or protein on which the activity of the target is dependent.

[0037] As used herein, the terms “effective amount" and "amount effective” refer to an amount that is sufficient to achieve the desired result or to have an effect on an undesired condition. For example, a “therapeutically effective amount” refers to an amount that is sufficient to achieve the desired therapeutic result or to have an effect on undesired symptoms. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of a compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, single dose compositions can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual physician in the event of any contraindications. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products. In further various aspects, a preparation can be administered in a "prophylactically effective amount"; that is, an amount effective for prevention of a disease or condition.

[0038] As used herein, “kit” means a collection of at least two components constituting the kit. Together, the components constitute a functional unit for a given purpose. Individual member components may be physically packaged together or separately. For example, a kit comprising an instruction for using the kit may or may not physically include the instruction with other individual member components. Instead, the instruction can be supplied as a separate member component, either in a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as recorded presentation.ATTORNEY DOCKET NO.930307-2000

[0039] As used herein, “instruction(s)” means documents describing relevant materials or methodologies pertaining to a kit. These materials may include any combination of the following: background information, list of components and their availability information (purchase information, etc.), brief or detailed protocols for using the kit, trouble-shooting, references, technical support, and any other related documents. Instructions can be supplied with the kit or as a separate member component, either as a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as recorded presentation. Instructions can comprise one or multiple documents, and are meant to include future updates.

[0040] As used herein, the terms “therapeutic agent” include any synthetic or naturally occurring biologically active compound or composition of matter which, when administered to an organism (human or nonhuman animal), induces a desired pharmacologic, immunogenic, and / or physiologic effect by local and / or systemic action. The term therefore encompasses those compounds or chemicals traditionally regarded as drugs, vaccines, and biopharmaceuticals including molecules such as proteins, peptides, hormones, nucleic acids, gene constructs and the like. Examples of therapeutic agents are described in well-known literature references such as the Merck Index (14th edition), the Physicians' Desk Reference (64th edition), and The Pharmacological Basis of Therapeutics (12th edition), and they include, without limitation, medicaments; vitamins; mineral supplements; substances used for the treatment, prevention, diagnosis, cure or mitigation of a disease or illness; substances that affect the structure or function of the body, or pro-drugs, which become biologically active or more active after they have been placed in a physiological environment. For example, the term "therapeutic agent" includes compounds or compositions for use in all of the major therapeutic areas including, but not limited to, adjuvants; anti-infectives such as antibiotics and antiviral agents; analgesics and analgesic combinations, anorexics, anti-inflammatory agents, anti-epileptics, local and general anesthetics, hypnotics, sedatives, antipsychotic agents, neuroleptic agents, antidepressants, anxiolytics, antagonists, neuron blocking agents, anticholinergic and cholinomimetic agents, antimuscarinic and muscarinic agents, antiadrenergics, antiarrhythmics, antihypertensive agents, hormones, and nutrients, antiarthritics, antiasthmatic agents, anticonvulsants, antihistamines, antinauseants, antineoplastics, antipruritics, antipyretics; antispasmodics, cardiovascular preparations (including calcium channel blockers, beta-blockers, beta-agonists and antiarrythmics), antihypertensives, diuretics, vasodilators; central nervous system stimulants; cough and cold preparations; decongestants; diagnostics; hormones; bone growth stimulants and bone resorption inhibitors;ATTORNEY DOCKET NO.930307-2000 immunosuppressives; muscle relaxants; psychostimulants; sedatives; tranquilizers; proteins, peptides, and fragments thereof (whether naturally occurring, chemically synthesized or recombinantly produced); and nucleic acid molecules (polymeric forms of two or more nucleotides, either ribonucleotides (RNA) or deoxyribonucleotides (DNA) including both double- and single-stranded molecules, gene constructs, expression vectors, antisense molecules and the like), small molecules (e.g., doxorubicin) and other biologically active macromolecules such as, for example, proteins and enzymes. The agent may be a biologically active agent used in medical, including veterinary, applications and in agriculture, such as with plants, as well as other areas. The term therapeutic agent also includes without limitation, medicaments; vitamins; mineral supplements; substances used for the treatment, prevention, diagnosis, cure or mitigation of disease or illness; or substances which affect the structure or function of the body; or pro- drugs, which become biologically active or more active after they have been placed in a predetermined physiological environment.

[0041] As used herein, “IC50,” is intended to refer to the concentration of a substance (e.g., acompound or a drug) that is required for 50% modulation of a biological process, or component of a process. For example, IC50refers to the half maximal (50%) inhibitory concentration (IC) of a substance as determined in a suitable assay. For example, an IC50for ANGPTL4 can be determined in an in vitro using purified or isolated ANGPTL4, or alternatively in an in vitro cell- based assay system. Frequently, assays, including suitable assays for ANGPTL4 modulation, make use of a suitable cell-line, e.g. a cell line that either expresses endogenously a target of interest, has been transfected with a suitable expression vector that directs expression of a recombinant form of the target such as ANGPTL4, or is a cell-line derived from a cancer that can be associated with ANGPTL4 over-expression. For example, the IC50for ANGPTL4 can be determined using cell-lines such as PANC-1, ASPC 1, BXPC-3, or MIA-PaCa-2 cells, and determined the effect of a compound on cell proliferation using an assay for cellular proliferation, e.g., a luminescent-based assay such as the Cell Titer-Glo® (Promega Corporation, Madison, Wisconsin).

[0042] The term “pharmaceutically acceptable” describes a material that is not biologically or otherwise undesirable, i.e., without causing an unacceptable level of undesirable biological effects or interacting in a deleterious manner.

[0043] As used herein, the term “pharmaceutically acceptable carrier” refers to sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterileATTORNEY DOCKET NO.930307-2000 powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid and the like. It can also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents, such as aluminum monostearate and gelatin, which delay absorption. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable media just prior to use. Suitable inert carriers can include sugars such as lactose. Desirably, at least 95% by weight of the particles of the active ingredient have an effective particle size in the range of 0.01 to 10 micrometers.

[0044] A residue of a chemical species, as used in the specification and concluding claims, refers to the moiety that is the resulting product of the chemical species in a particular reaction scheme or subsequent formulation or chemical product, regardless of whether the moiety is actually obtained from the chemical species. Thus, an ethylene glycol residue in a polyester refers to one or more -OCH2CH2O- units in the polyester, regardless of whether ethylene glycol was used to prepare the polyester. Similarly, a sebacic acid residue in a polyester refers to one or more -CO(CH2)8CO- moieties in the polyester, regardless of whether the residue is obtained by reacting sebacic acid or an ester thereof to obtain the polyester.

[0045] As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclicATTORNEY DOCKET NO.930307-2000 and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. Also, the terms “substitution” or “substituted with” include the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. It is also contemplated that, in certain aspects, unless expressly indicated to the contrary, individual substituents can be further optionally substituted (i.e., further substituted or unsubstituted).

[0046] The term “aliphatic” or “aliphatic group,” as used herein, denotes a hydrocarbon moiety that may be straight-chain (i.e., unbranched), branched, or cyclic (including fused, bridging, and spirofused polycyclic) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic. Unless otherwise specified, aliphatic groups contain 1-20 carbon atoms. Aliphatic groups include, but are not limited to, linear or branched, alkyl, alkenyl, and alkynyl groups, and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0047] The term “alkyl” as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. The alkyl group can be cyclic or acyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein. A “lower alkyl” group is an alkyl group containing from one to six (e.g., from one to four) carbon atoms. The term alkyl group can also be a C1 alkyl, C1-C2 alkyl, C1-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C8 alkyl, C1-C9 alkyl, C1-C10 alkyl, and the like up to and including a C1-C24 alkyl.

[0048] Throughout the specification “alkyl” is generally used to refer to both unsubstitutedATTORNEY DOCKET NO.930307-2000 alkyl groups and substituted alkyl groups; however, substituted alkyl groups are also specifically referred to herein by identifying the specific substituent(s) on the alkyl group. For example, the term “halogenated alkyl” or “haloalkyl” specifically refers to an alkyl group that is substituted with one or more halide, e.g., fluorine, chlorine, bromine, or iodine. Alternatively, the term “monohaloalkyl” specifically refers to an alkyl group that is substituted with a single halide, e.g. fluorine, chlorine, bromine, or iodine. The term “polyhaloalkyl” specifically refers to an alkyl group that is independently substituted with two or more halides, i.e. each halide substituent need not be the same halide as another halide substituent, nor do the multiple instances of a halide substituent need to be on the same carbon. The term “alkoxyalkyl” specifically refers to an alkyl group that is substituted with one or more alkoxy groups, as described below. The term “aminoalkyl” specifically refers to an alkyl group that is substituted with one or more amino groups. The term “hydroxyalkyl” specifically refers to an alkyl group that is substituted with one or more hydroxy groups. When “alkyl” is used in one instance and a specific term such as “hydroxyalkyl” is used in another, it is not meant to imply that the term “alkyl” does not also refer to specific terms such as “hydroxyalkyl” and the like.

[0049] This practice is also used for other groups described herein. That is, while a term such as “cycloalkyl” refers to both unsubstituted and substituted cycloalkyl moieties, the substituted moieties can, in addition, be specifically identified herein; for example, a particular substituted cycloalkyl can be referred to as, e.g., an “alkylcycloalkyl.” Similarly, a substituted alkoxy can be specifically referred to as, e.g., a “halogenated alkoxy,” a particular substituted alkenyl can be, e.g., an “alkenylalcohol,” and the like. Again, the practice of using a general term, such as “cycloalkyl,” and a specific term, such as “alkylcycloalkyl,” is not meant to imply that the general term does not also include the specific term.

[0050] The terms “alkoxy” and “alkoxyl” as used herein to refer to an alkyl or cycloalkyl group bonded through an ether linkage; that is, an “alkoxy” group can be defined as —OA1where A1is alkyl or cycloalkyl as defined above. “Alkoxy” also includes polymers of alkoxy groups as just described; that is, an alkoxy can be a polyether such as —OA1—OA2or —OA1—(OA2)a—OA3, where “a” is an integer of from 1 to 200 and A1, A2, and A3are alkyl and / or cycloalkyl groups.

[0051] The term “aromatic group” as used herein refers to a ring structure having cyclic clouds of delocalized π electrons above and below the plane of the molecule, where the π clouds contain (4n+2) π electrons. A further discussion of aromaticity is found in Morrison and Boyd, Organic Chemistry, (5th Ed., 1987), Chapter 13, entitled “Aromaticity,” pages 477-497, incorporated hereinATTORNEY DOCKET NO.930307-2000 by reference. The term “aromatic group” is inclusive of both aryl and heteroaryl groups.

[0052] The term “aryl” as used herein is a group that contains any carbon-based aromatic group including, but not limited to, benzene, naphthalene, phenyl, biphenyl, anthracene, and the like. The aryl group can be substituted or unsubstituted. The aryl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, ─NH2, carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol as described herein. The term “biaryl” is a specific type of aryl group and is included in the definition of “aryl.” In addition, the aryl group can be a single ring structure or comprise multiple ring structures that are either fused ring structures or attached via one or more bridging groups such as a carbon-carbon bond. For example, biaryl to two aryl groups that are bound together via a fused ring structure, as in naphthalene, or are attached via one or more carbon-carbon bonds, as in biphenyl.

[0053] The terms “amine” or “amino” as used herein are represented by the formula —NA1A2, where A1and A2can be, independently, hydrogen or alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. A specific example of amino is ─NH2.

[0054] The term “alkylamino” as used herein is represented by the formula —NH(-alkyl) or —N(-alkyl)2, where alkyl is a described herein. Representative examples include, but are not limitedto, methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, isobutylamino group, (sec-butyl)amino group, (tert-butyl)amino group, pentylamino group, isopentylamino group, (tert-pentyl)amino group, hexylamino group, dimethylamino group, diethylamino group, dipropylamino group, diisopropylamino group, dibutylamino group, diisobutylamino group, di(sec-butyl)amino group, di(tert-butyl)amino group, dipentylamino group, diisopentylamino group, di(tert-pentyl)amino group, dihexylamino group, N-ethyl-N-methylamino group, N-methyl-N-propylamino group, N-ethyl-N-propylamino group and the like.

[0055] The term “nitro” as used herein is represented by the formula —NO2.

[0056] The term “nitrile” or “cyano” as used herein is represented by the formula —CN.

[0057] The term “sulfo-oxo” as used herein is represented by the formulas —S(O)A1, — S(O)2A1, —OS(O)2A1, or —OS(O)2OA1, where A1can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. Throughout thisATTORNEY DOCKET NO.930307-2000 specification “S(O)” is a short hand notation for S=O. The term “sulfonyl” is used herein to refer to the sulfo-oxo group represented by the formula —S(O)2A1, where A1can be hydrogen or an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “sulfone” as used herein is represented by the formula A1S(O)2A2, where A1and A2can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein. The term “sulfoxide” as used herein is represented by the formula A1S(O)A2, where A1and A2can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.

[0058] The term “thiol” as used herein is represented by the formula —SH.

[0059] “A1,” “A2,” …“An”; “L1,” “L2,” …“Ln”; and “R1,” “R2,” …“Rn”; and the like, where n is an integer, are used herein as generic symbols to represent various specific substituents and can, independently, possess one or more of the groups listed above as defined within this disclosure. These symbols can be any substituent, not limited to those disclosed herein, and when they are defined to be certain substituents in one instance, they can, in another instance, be defined as some other substituents. For example, if R1is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group can optionally be substituted with a hydroxyl group, an alkoxy group, an alkyl group, a halide, and the like. Depending upon the groups that are selected, a first group can be incorporated within second group or, alternatively, the first group can be pendant (i.e., attached) to the second group. For example, with the phrase “an alkyl group comprising an amino group,” the amino group can be incorporated within the backbone of the alkyl group. Alternatively, the amino group can be attached to the backbone of the alkyl group. The nature of the group(s) that is (are) selected will determine if the first group is embedded or attached to the second group.

[0060] The term “organic residue” defines a carbon containing residue, i.e., a residue comprising at least one carbon atom, and includes but is not limited to the carbon-containing groups, residues, or radicals defined hereinabove. Organic residues can contain various heteroatoms, or be bonded to another molecule through a heteroatom, including oxygen, nitrogen, sulfur, phosphorus, or the like. Examples of organic residues include but are not limited alkyl or substituted alkyls, alkoxy or substituted alkoxy, mono or di-substituted amino, amide groups, etc. Organic residues can preferably comprise 1 to 18 carbon atoms, 1 to 15, carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In an aspect, an organic residue can comprise 2 to 18 carbon atoms, 2 to 15, carbon atoms, 2 to 12 carbon atoms, 2 to 8 carbon atoms, 2 to 4 carbon atoms, or 2 to 4 carbon atoms.ATTORNEY DOCKET NO.930307-2000

[0061] A very close synonym of the term “residue” is the term “radical,” which as used in the specification and concluding claims, refers to a fragment, group, or substructure of a molecule described herein, regardless of how the molecule is prepared. For example, a 2,4- thiazolidinedione radical in a particular compound has the structure: O NHwhether thiazolidinedione is used to prepare the compound. In some embodiments the radical (for example an alkyl) can be further modified (i.e., substituted alkyl) by having bonded thereto one or more “substituent radicals.” The number of atoms in a given radical is not critical to the present disclosure unless it is indicated to the contrary elsewhere herein.

[0062] “Organic radicals,” as the term is defined and used herein, contain one or more carbon atoms. An organic radical can have, for example, 1-26 carbon atoms, 1-18 carbon atoms, 1-12 carbon atoms, 1-8 carbon atoms, 1-6 carbon atoms, or 1-4 carbon atoms. In an aspect, an organic radical can have 2-26 carbon atoms, 2-18 carbon atoms, 2-12 carbon atoms, 2-8 carbon atoms, 2-6 carbon atoms, or 2-4 carbon atoms. Organic radicals often have hydrogen bound to at least some of the carbon atoms of the organic radical. One example, of an organic radical that comprises no inorganic atoms is a 5, 6, 7, 8-tetrahydro-2-naphthyl radical. In some embodiments, an organic radical can contain 1-10 inorganic heteroatoms bound thereto or therein, including halogens, oxygen, sulfur, nitrogen, phosphorus, and the like. Examples of organic radicals include but are not limited to an alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, mono- substituted amino, di-substituted amino, acyloxy, cyano, carboxy, carboalkoxy, alkylcarboxamide, substituted alkylcarboxamide, dialkylcarboxamide, substituted dialkylcarboxamide, alkylsulfonyl, alkylsulfinyl, thioalkyl, thiohaloalkyl, alkoxy, substituted alkoxy, haloalkyl, haloalkoxy, aryl, substituted aryl, heteroaryl, heterocyclic, or substituted heterocyclic radicals, wherein the terms are defined elsewhere herein. A few non-limiting examples of organic radicals that include heteroatoms include alkoxy radicals, trifluoromethoxy radicals, acetoxy radicals, dimethylamino radicals and the like.

[0063] Compounds described herein can contain one or more double bonds and, thus, potentially give rise to cis / trans (E / Z) isomers, as well as other conformational isomers. Unless stated to the contrary, the disclosure includes all such possible isomers, as well as mixtures ofATTORNEY DOCKET NO.930307-2000 such isomers.

[0064] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer and diastereomer, and a mixture of isomers, such as a racemic or scalemic mixture. Compounds described herein can contain one or more asymmetric centers and, thus, potentially give rise to diastereomers and optical isomers. Unless stated to the contrary, the present disclosure includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof. Mixtures of stereoisomers, as well as isolated specific stereoisomers, are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.

[0065] Many organic compounds exist in optically active forms having the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are non-superimposable mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture. Many of the compounds described herein can have one or more chiral centers and therefore can exist in different enantiomeric forms. If desired, a chiral carbon can be designated with an asterisk (*). When bonds to the chiral carbon are depicted as straight lines in the disclosed formulas, it is understood that both the (R) and (S) configurations of the chiral carbon, and hence both enantiomers and mixtures thereof, are embraced within the formula. As is used in the art, when it is desired to specify the absolute configuration about a chiral carbon, one of the bonds to the chiral carbon can be depicted as a wedge (bonds to atoms above the plane) and the other can be depicted as a series or wedge of short parallel lines is (bonds to atoms below the plane). The Cahn-Inglod-Prelog system can be used to assign the (R) or (S) configuration to a chiral carbon.

[0066] Compounds described herein comprise atoms in both their natural isotopic abundanceATTORNEY DOCKET NO.930307-2000 and in non-natural abundance. Disclosed compounds can be isotopically-labeled or isotopically- substituted compounds identical to those described, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine and chlorine, such as2H,3H,13C,14C,15N,18O,17O,35S,18F, and36Cl, respectively. Compounds further comprise prodrugs thereof and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this disclosure. Certain isotopically-labeled compounds of the present disclosure, for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e.,3H, and carbon-14, i.e.,14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e.,2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds of the present disclosure and prodrugs thereof can generally be prepared by carrying out the procedures below, by substituting a readily available isotopically labeled reagent for a non- isotopically labeled reagent.

[0067] The compounds described in the disclosure can be present as a solvate. In some cases, the solvent used to prepare the solvate is an aqueous solution, and the solvate is then often referred to as a hydrate. The compounds can be present as a hydrate, which can be obtained, for example, by crystallization from a solvent or from aqueous solution. In this connection, one, two, three or any arbitrary number of solvent or water molecules can combine with the compounds according to the disclosure to form solvates and hydrates. Unless stated to the contrary, the disclosure includes all such possible solvates.

[0068] The term “co-crystal” means a physical association of two or more molecules which owe their stability through non-covalent interaction. One or more components of this molecular complex provide a stable framework in the crystalline lattice. In certain instances, the guest molecules are incorporated in the crystalline lattice as anhydrates or solvates, see e.g. “Crystal Engineering of the Composition of Pharmaceutical Phases. Do Pharmaceutical Co-crystals Represent a New Path to Improved Medicines?” Almarasson, O., et al., The Royal Society of Chemistry, 1889-1896, 2004. Examples of co-crystals include p-toluenesulfonic acid and benzenesulfonic acid.ATTORNEY DOCKET NO.930307-2000

[0069] The term “nanoparticle” means a particle that has an average particle size of less than about 500 nm, as measured by dynamic light scattering (DLS), e.g., less than about 400 nm, less than about 300 nm, less than about 200 nm, less than about 190 nm, or from about 100 to about 190 nm or from about 100 to about 150 nm. In some aspects, average particle size, when measured by DLS, can represent the hydrodynamic diameter of the particle. DLS measurements typically have a margin of error around ±10%.

[0070] A “pH-responsive polymer carrier” is a polymer that when exposed to a certain pH, the polymer carrier is destabilized and the pH-responsive nanoparticle releases astemizole. In one aspect, the pH-responsive polymer carrier becomes destabilized at a pH from about 2 to about 7.5. In another aspect, the pH-responsive polymer carrier becomes destabilized at a pH of about 7.4.

[0071] It is also appreciated that certain compounds described herein can be present as an equilibrium of tautomers. For example, ketones with an α-hydrogen can exist in an equilibrium of the keto form and the enol form. OOHO OHform Likewise, amides with an N-hydrogen can exist in an equilibrium of the amide form and the imidic acid form. Unless stated to the contrary, the disclosure includes all such possible tautomers.

[0072] It is known that chemical substances form solids which are present in different states of order which are termed polymorphic forms or modifications. The different modifications of a polymorphic substance can differ greatly in their physical properties. The compounds according to the disclosure can be present in different polymorphic forms, with it being possible for particular modifications to be metastable. Unless stated to the contrary, the disclosure includes all such possible polymorphic forms.

[0073] In some aspects, a structure of a compound can be represented by a formula: ,ATTORNEY DOCKET NO.930307-2000 which is understood to be equivalent to a formula: Rn(a)Rn(b)(c), wherein n is typically an integer.to represent five independent substituents, Rn(a), Rn(b), Rn(c), Rn(d), and Rn(e). By “independent substituents,” it is meant that each R substituent can be independently defined. For example, if in one instance Rn(a)is halogen, then Rn(b)is not necessarily halogen in that instance.

[0074] As used herein, “astemizole” refers to a compound having a structure represented by the formla: O .compositions, and components disclosed herein can be obtained commercially or readily synthesized using techniques generally known to those of skill in the art. For example, the starting materials and reagents used in preparing disclosed compounds and compositions are either available from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), Acros Organics (Morris Plains, N.J.), Fisher Scientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.) or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March’s Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

[0076] Unless otherwise expressly stated, it is in no way intended that any method set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, whereATTORNEY DOCKET NO.930307-2000 a method claim does not actually recite an order to be followed by its steps or it is not otherwise specifically stated in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non- express basis for interpretation, including: matters of logic with respect to arrangement of steps or operational flow; plain meaning derived from grammatical organization or punctuation; and the number or type of embodiments described in the specification.

[0077] Disclosed are the components to be used to prepare the compositions of the disclosure as well as the compositions themselves to be used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds cannot be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed and a number of modifications that can be made to a number of molecules including the compounds are discussed, specifically contemplated is each and every combination and permutation of the compound and the modifications that are possible unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited each is individually and collectively contemplated meaning combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considered disclosed. Likewise, any subset or combination of these is also disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the compositions of the disclosure. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the disclosure.

[0078] It is understood that the compositions disclosed herein have certain functions. Disclosed herein are certain structural requirements for performing the disclosed functions, and it is understood that there are a variety of structures that can perform the same function that are related to the disclosed structures, and that these structures will typically achieve the same result.

[0079] In an aspect, the disclosure pertains to compounds useful as inhibitors of ANGPTL4. In an aspect, the compounds of the disclosure are useful in the treatment of disorders of uncontrolled cellular proliferation, e.g., a cancer, associated with an ANGPTL4 dysfunction, e.g.,ATTORNEY DOCKET NO.930307-2000 aberrant expression levels of ANGPTL4, and other diseases in which an ANGPTL4 dysfunction is involved.

[0080] It is contemplated that each disclosed derivative can be optionally further substituted. It is also contemplated that any one or more derivative can be optionally omitted from the disclosure. It is understood that a disclosed compound can be provided by the disclosed methods. It is also understood that disclosed compounds can be employed in the disclosed methods of using. pH-Responsive Nanoparticles Composed of Astemizole

[0081] The pH-responsive nanoparticles described herein facilitate the release of astemizole to effectively release astemizole locally, preventing or reducing cancer cell growth, while minimizing effects on healthy cells. The compositions comprise a pH-responsive nanoparticle having a core comprising astemizole or a pharmaceutically acceptable salt thereof encapsulated by a pH-responsive polymer carrier. The pH-responsive nanoparticles can be administred alone or in combination with other other pharmaceutical compounds such as, for example, anticancer agents.

[0082] Described herein are methods of making the pH-responsive nanoparticles that can be useful in the treatment of disorders of uncontrolled cellular proliferation associated with an ANGPTL4 dysfunction and other diseases in which ANGPTL4 is involved. In an aspect, the disclosure pertains to the disclosed synthetic manipulations. In an aspect, the disclosed compounds comprise the products of the synthetic methods described herein. In an aspect, the disclosed compounds comprise a compound produced by a synthetic method described herein. In an aspect, the disclosure comprises a pharmaceutical composition comprising a therapeutically effective amount of the product of the disclosed methods and a pharmaceutically acceptable carrier. In an aspect, the disclosure comprises a method for manufacturing a medicament comprising combining at least one compound of any of disclosed compounds or at least one product of the disclosed methods with a pharmaceutically acceptable carrier or diluent.

[0083] The compounds of this disclosure can be prepared by employing reactions as shown in the disclosed schemes, in addition to other standard manipulations that are known in the literature, exemplified in the experimental sections or clear to one skilled in the art. The following examples are provided so that the disclosure might be more fully understood, are illustrative only, and should not be construed as limiting. For clarity, examples having a fewer substituent can beATTORNEY DOCKET NO.930307-2000 shown where multiple substituents are allowed under the definitions disclosed herein.

[0084] In one aspect, the pH-responsive nanoparticles can be prepared by a nonsolvent- induced phase separation (nanoprecipitation) method using a selective solvent (e.g., DMSO) and as a nonselective solvent (e.g., PBS at pH 7.4). Initially, the pH-responsive polymer carrier and astemizole are dissolved in the selective solvent, and the resultant solution is dialyzed against the nonselective solvent. The dialyzed suspension can then be filtered to remove unencapsulated stemizole to obtain the pH-responsive nanoparticles.

[0085] It is contemplated that each disclosed method can further comprise additional steps, manipulations, and / or components. It is also contemplated that any one or more step, manipulation, and / or component can be optionally omitted from the disclosure.

[0086] In one aspect, the pH-responsive polymer carrier comprises a copolymer of poly(ethylene glycol) (PEG) and a poly(carbonate). In this aspect, pH-responsive polymer carrier comprises a copolymer of poly(ethylene glycol) (PEG) and a poly(carbonate). The copolymer can be a random copolymer of PEG and the poly(carbonate) but in some aspects the copolymer is a block copolymer featuring a PEG block and a block of the polycarbonate. The copolymers of poly(ethylene glycol) (PEG) and a poly(carbonate) disclosed in US Publication No.2022 / 0378711, which is incorporated by reference for its teachings regarding the copolymers, can be used as the pH-responsive polymer carrier as described herein.

[0087] The molecular weight of the poly(ethylene glycol) (PEG) and a poly(carbonate) can vary. In some aspects, the copolymer has a number-average molecular weight (Ma) as determined by gel-permeation chromatography (GPC) ranging from about 1,000 Daltons to about 50,000 Daltons. In a further aspect, the Mnof the copolymer ranges from about 5,000 Daltons to about 50,000 Daltons, e.g., about 10,000 Daltons to about 50,000 Daltons, about 15,000 Daltons to about 50,000 Daltons, or about 20,000 Daltons to about 50,000 Daltons. In a still further aspect, the Mnof the copolymer ranges from about 20,000 Daltons to about 40,000 Daltons, e.g., about 20,000 Daltons, about 25,000 Daltons, about 30,000 Daltons, or about 45,000 Daltons. In some aspects, when the copolymer is a block copolymer, the PEG block has a molecular weight ranging from about 5,000 Daltons to about 20,000 Daltons, while the poly(carbonate) block has a molecular weight of about 15,000 Daltons.

[0088] In one aspect, the copolymer of poly(ethylene glycol) (PEG) and a poly(carbonate) has a structure represented by the formulaATTORNEY DOCKET NO.930307-2000wherein R1is C1-C4alkoxy, R2is C1-C12alkyl, C1-C12alkylamino, C2-C12dialkylamino, C3- C12cycloalkyl, or C2-C12heterocycloalkyl, and n and m are independently integers ranging from 10 to 300.

[0089] In another aspect, the pH-responsive polymer carrier has a structural unit selected from the group consisting of .ATTORNEY DOCKET NO.930307-2000

[0090] In another aspect, the pH-responsive polymer carrier is II

[0091] x is a single integer greater than 500 or a single integer greater than 1,000. In other aspects, x is a single integer between 1 and 100, 1 and 1000, 1 and 10,000 or 1 and 1,000,000. In other aspects, x is a single integer between 500 and a million. In yet other aspects, x is a single integer between 100 and 100,000.

[0092] In another aspect, n in structure II is a large number that varies depending on polymer polydispersity. In one aspect, n is a single integer greater than 1. In other aspects, n is a single integer greater than 500 or a single integer greater than 1,000. In some aspects, n is a single integer between 1 and 100, 1 and 1000, 1 and 10,000 or 1 and 1,000,000. In other aspects n is a single integer between 500 and a million. In yet other aspects, n is a single integer between 100 and 100,000. The polymers disclosed in US Patent No. 9,687,553, which is incorporated by reference for its teachings regarding the polymers, can be used as the pH-responsive polymer carrier as described herein.

[0093] In another aspect, the pH-responsive polymer carrier comprises (a) a first polymer comprising a polysaccharide; and a second polymer comprising an alkylaminoalkyl ester or methacrylic acid grafted to the first polymer. The polymers disclosed in US Patent No. 11,155,664, which is incorporated by reference for its teachings regarding the polymers, can be used as the pH-responsive polymer carrier as described herein. In one aspect, the first polymer comprises a polysaccharide, wherein the polysaccharide has a degree of substitution between 0.5 and 3 per monomeric unit of the polysaccharide, wherein the first polymer is a polyhydroxyl polymer; the second polymer comprises a monomer selected from the group consisting ofATTORNEY DOCKET NO.930307-2000 methacrylic acid and diethylaminoethyl methacrylic acid, wherein the second polymer is grafted to the first polymer; and a polyethoxylated sorbitan-based moiety covalently bound to the second polymer.

[0094] In another aspect, the pH-responsive polymer carrier comprises a block copolymer comprising a hydrophilic polymer segment and a hydrophobic polymer segment, wherein the hydrophilic polymer segment comprises a polymer selected from the group consisting of: poly(ethylene oxide) (PEO), poly(methacrylate phosphatidyl choline) (MPC), and polyvinylpyrrolidone (PVP), wherein the hydrophilic polymer segment has from 100 to 130 monomer units, wherein the hydrophobic polymer segment comprises wherein R′ is H or CH3,wherein R is NR1R2, wherein R1and R2are n-butyl, wherein n is 1 to about 10, and wherein x is about 20 to about 200 in total.

[0095] The polymers disclosed in US Publication No.2021 / 0198410, which is incorporated by reference for its teachings regarding the polymers, can be used as the pH-responsive polymer carrier as described herein. Pharmaceutical Compositions

[0096] In one aspect, described herein are pharmaceutical compositions comprising the pH- responsive nanoparticles described herein. That is, a pharmaceutical composition can be provided comprising an effective amount of astemizole in the pH-responsive nanoparticle, and a pharmaceutically acceptable carrier.ATTORNEY DOCKET NO.930307-2000

[0097] In an aspect, the effective amount of the astemizole in the pH-responsive nanoparticle is a therapeutically effective amount. In an aspect, the effective amount is a prophylactically effective amount of astemizole.

[0098] In one aspect, the pharmaceutical compositions described herein exhibit modulation of ANGPTL4 activity with a binding affinity Kiof less than about 50 µM, 25 µM, 10 µM, 5 µM, 1 µM, 500 nM, 250 nM, 100 nM, 50 nM, 25 nM, 10 nM, 5 nM, or 1 nM. In an aspect, modulation of ANGPTL4 activity is inhibition of ANGPTL4 activity.

[0099] In one aspect, the pharmaceutical compositions described herein exhibit inhibition of cellular proliferation with an IC50of less than about 50 µM, 25 µM, 10 µM, 5 µM, 1 µM, 500 nM, 250 nM, 100 nM, 50 nM, 25 nM, 10 nM, 5 nM, or 1 nM.

[0100] In one aspect, the pharmaceutical composition is used to treat a mammal. In an aspect, the mammal is a human. In an aspect, the mammal has been diagnosed with a need for treatment of the disorder prior to the administering step. In an aspect, the mammal has been identified to be in need of treatment of the disorder. In an aspect, the pharmaceutical composition is used to treat a disorder of uncontrolled cellular proliferation. In an aspect, the disorder is associated with an ANGPTL4 dysfunction.

[0101] In one aspect, the pharmaceutical composition is used to treat a cancer. In an aspect, the pharmaceutical composition is used to treat a cancer such as a pancreatic cancer, breast cancer, renal cancer, gastric cancer, colorectal cancer, lymphoma, cancers of the brain, genitourinary tract cancer, lymphatic system cancer, stomach cancer, larynx cancer, lung cancer, and malignant melanoma. In an aspect, the pharmaceutical composition is used to treat a pancreatic cancer.

[0102] In one aspect, the pharmaceutical composition is used to treat a gemcitabine-resistant cancer. In an aspect, the pharmaceutical composition is used to treat a gemcitabine-resistant cancer such as a gemcitabine-resistant pancreatic cancer, gemcitabine-resistant breast cancer, gemcitabine-resistant renal cancer, gemcitabine-resistant gastric cancer, gemcitabine-resistant colorectal cancer, gemcitabine-resistant lymphoma, gemcitabine-resistant cancers of the brain, gemcitabine-resistant genitourinary tract cancer, gemcitabine-resistant lymphatic system cancer, gemcitabine-resistant stomach cancer, gemcitabine-resistant larynx cancer, gemcitabine- resistant lung cancer, and gemcitabine-resistant melanoma.In an aspect, the pharmaceutical composition is used to treat a gemcitabine-resistant pancreatic cancer.ATTORNEY DOCKET NO.930307-2000

[0103] In certain aspects, the disclosed pharmaceutical compositions comprise astemizole (including pharmaceutically acceptable salt(s) thereof) as an active ingredient, a pharmaceutically acceptable carrier, and, optionally, other therapeutic ingredients or adjuvants. The instant compositions include those suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0104] As used herein, the term “pharmaceutically acceptable salts” refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When a compound is acidic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (-ic and -ous), ferric, ferrous, lithium, magnesium, manganese (-ic and -ous), potassium, sodium, zinc and the like salts. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines such as naturally occurring and synthesized substituted amines. Other pharmaceutically acceptable organic non-toxic bases from which salts can be formed include ion exchange resins such as, for example, arginine, betaine,caffeine, choline, N,N’-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

[0105] As used herein, the term “pharmaceutically acceptable non-toxic acids,” includes inorganic acids, organic acids, and salts prepared therefrom, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like. Preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.ATTORNEY DOCKET NO.930307-2000

[0106] In practice, the pH-responsive nanoparticles described herein can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present disclosure can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compounds of the disclosure, and / or pharmaceutically acceptable salt(s) thereof, can also be administered by controlled release means and / or delivery devices. The compositions can be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

[0107] Thus, the pharmaceutical compositions of this disclosure can include a pharmaceutically acceptable carrier and the pH-responsive nanoparticles described herein. The pH-responsive nanoparticles described herein can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

[0108] The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0109] In preparing the compositions for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solidATTORNEY DOCKET NO.930307-2000 pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques

[0110] A tablet containing the pH-responsive nanoparticles described herein can be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.

[0111] In one aspect, the pharmaceutical compositions of the present disclosure comprise the pH-responsive nanoparticles described herein as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents or adjuvants. The instant compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0112] Pharmaceutical compositions of the present disclosure suitable for parenteral administration can be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

[0113] Pharmaceutical compositions of the present disclosure suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.ATTORNEY DOCKET NO.930307-2000

[0114] Pharmaceutical compositions of the present disclosure can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, mouth washes, gargles, and the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations can be prepared, utilizing a compound of the disclosure, or pharmaceutically acceptable salts thereof, via conventional processing methods. As an example, a cream or ointment is prepared by mixing hydrophilic material and water, together with about 5 wt% to about 10 wt% of the compound, to produce a cream or ointment having a desired consistency.

[0115] Pharmaceutical compositions of this disclosure can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories can be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

[0116] In addition to the aforementioned carrier ingredients, the pharmaceutical formulations described above can include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing a compound of the disclosure, and / or pharmaceutically acceptable salts thereof, can also be prepared in powder or liquid concentrate form.

[0117] In the treatment conditions which require modulation of ANGPTL4 activity, an appropriate dosage level of astemizole will generally be about 0.01 to 500 mg per kg patient body weight per day and can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg / kg per day; more preferably 0.5 to 100 mg / kg per day. A suitable dosage level can be about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5.0 or 5.0 to 50 mg / kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient for the symptomatic adjustment of the dosage of the patient to be treated. The pH-responsive nanoparticles described herein can be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosing regimen can be adjusted to provide theATTORNEY DOCKET NO.930307-2000 optimal therapeutic response.

[0118] It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors. Such factors include the age, body weight, general health, sex, and diet of the patient. Other factors include the time and route of administration, rate of excretion, drug combination, and the type and severity of the particular disease undergoing therapy.

[0119] The present disclosure is further directed to a method for the manufacture of a medicament for modulating ANGPTL4 activity (e.g., treatment of one or more disorder of uncontrolled cellular proliferation associated with ANGPTL4 dysfunction) in mammals (e.g., humans) comprising combining one or more disclosed compounds, products, or compositions with a pharmaceutically acceptable carrier or diluent.

[0120] It is understood that the disclosed pharmaceutical compositions can be prepared from disclosed compounds. It is also understood that the disclosed pharmaceutical compositions can be employed in the disclosed methods of using. Methods of Use

[0121] The pH-responsive nanoparticles described herein are useful for treating, preventing, ameliorating, controlling or reducing the risk of a variety of disorders associated with uncontrolled cellular proliferation. For example, a treatment can include modulation of ANGPTL4 to an extent effective to a cancer. Thus, a disorder can be associated with ANGPTL4 activity. Thus, provided is a method of treating or preventing a disorder in a subject comprising the step of administering to the subject the pH-responsive nanoparticles described herein; at least one disclosed pharmaceutical composition; and / or at least one disclosed product in a dosage and amount effective to treat the disorder in the subject.

[0122] In one aspect, the disclosure pertains to a method for the treatment of a disorder of uncontrolled cellular proliferation, e.g., a cancer, in a mammal comprising the step of administering to the mammal the pH-responsive nanoparticles described herein, a disclosed pharmaceutical composition, or a disclosed medicament.

[0123] In another aspect, the disclosure pertains to a method for the treatment of a gemcitabine-resistant cancer, in a mammal comprising the step of administering to the mammal the pH-responsive nanoparticles described herein, a disclosed pharmaceutical composition, or a disclosed medicament.ATTORNEY DOCKET NO.930307-2000

[0124] Also provided is a method for the treatment of one or more disorders, e.g., a cancer, associated with ANGPTL4 activity in a subject comprising the step of administering to the subject the pH-responsive nanoparticles described herein; or a disclosed medicament in a dosage and amount effective to treat the disorder in the subject.

[0125] In another aspect, the disclosure pertains to a method for modulation of ANGPTL4 activity in a mammal comprising the step of administering to the mammal an effective amount of the pH-responsive nanoparticles described herein.

[0126] In another aspect, the disclosure pertains to a method for modulation of ANGPTL4 activity in at least one cell, comprising the step of contacting the at least one cell with an effective amount the pH-responsive nanoparticles described herein; at least one disclosed pharmaceutical composition; or a disclosed medicament in a dosage and amount effective to inhibit ANGPTL4 activity in the at least one cell.

[0127] In another aspect, the disclosure pertains to a method for inhibiting metastasis in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles described herein.

[0128] In another aspect, the pH-responsive nanoparticles described herein have utility in treating a variety of cancers, e.g., a cancer associated with ANGPTL4 activity, including one or more of the following conditions or diseases: a pancreatic cancer, breast cancer, renal cancer, gastric cancer, colorectal cancer, lymphoma, cancers of the brain, genitourinary tract cancer, lymphatic system cancer, stomach cancer, larynx cancer, lung, pancreatic cancer, and malignant melanoma. In an aspect, the pH-responsive nanoparticles described herein have utility in treating a pancreatic cancer.

[0129] In another aspect, the pH-responsive nanoparticles described herein have utility in treating a variety of gemcitabine-resistant cancers, e.g., a cancer associated with ANGPTL4 activity, including one or more of the following gemcitabine-resistant cancers: gemcitabine- resistant pancreatic cancer, gemcitabine-resistant breast cancer, gemcitabine-resistant renal cancer, gemcitabine-resistant gastric cancer, gemcitabine-resistant colorectal cancer, gemcitabine-resistant lymphoma, gemcitabine-resistant cancers of the brain, gemcitabine- resistant genitourinary tract cancer, gemcitabine-resistant lymphatic system cancer, gemcitabine- resistant stomach cancer, gemcitabine-resistant larynx cancer, gemcitabine-resistant lung cancer, and gemcitabine-resistant melanoma. In another aspect, the pH-responsive nanoparticlesATTORNEY DOCKET NO.930307-2000 described herein have utility in treating a pancreatic cancer. In another aspect, the pH-responsive nanoparticles described herein have utility in treating a gemcitabine-resistant pancreatic cancer.

[0130] In another aspect, the cell is mammalian, for example, human. In an aspect, the cell has been isolated from a subject prior to the contacting step. In an aspect, contacting is via administration to a subject. In aspect, modulation is inhibition of ANGPTL4 activity.

[0131] In another aspect, the disclosure pertains to a method for modulation of ANGPTL4 activity in a subject comprising the step of administering to the subject the pH-responsive nanoparticles described herein in a dosage and amount effective to inhibit ANGPTL4 activity in the subject. In an aspect, the subject is mammalian, for example, human. In an aspect, the mammal has been diagnosed with a need for modulation of ANGPTL4 activity prior to the administering step. In an aspect, the mammal has been diagnosed with a need for modulation of ANGPTL4 activity prior to the administering step. In an aspect, the method further comprises the step of identifying a subject in need of modulation of ANGPTL4 activity.

[0132] In another aspect, the disclosure pertains to a method for the treatment of a cancer associated with an ANGPTL4 dysfunction, for example, a cancer associated with increased ANGPTL4 expression, in a mammal comprising the step of administering to the mammal the pH- responsive nanoparticles described herein in a dosage and amount effective to treat the disorder in the mammal. In an aspect, the mammal is a human. In an aspect, the mammal has been diagnosed with a need for treatment for the disorder prior to the administering step. In an aspect, the method further comprises the step of identifying a subject in need of treatment for the disorder.

[0133] In another aspect, the pH-responsive nanoparticles described herein can be used in combination with one or more other drugs in the treatment, prevention, control, amelioration, or reduction of risk of cancers or conditions for which disclosed compounds or the other drugs can have utility, where the combination of the drugs together are safer or more effective than either drug alone. Such other drug(s) can be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with the pH-responsive nanoparticles described herein.

[0134] When the pH-responsive nanoparticles described herein are used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and a disclosed compound is preferred. However, the combination therapy can also include therapies in which the pH-responsive nanoparticles described herein and one or moreATTORNEY DOCKET NO.930307-2000 other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, disclosed compounds and the other active ingredients can be used in lower doses than when each is used singly.

[0135] In one aspect, the pH-responsive nanoparticles described herein are administered to the mammal prior to the administration of the drug (e.g., an anticancer agent). In another aspect, the pH-responsive nanoparticles are administered to mammal at least 12 hours prior to the administration of the drug. In another aspect, the pH-responsive nanoparticles are administered to mammal at least 12 hours to up to 96 hours prior to the administration of the anticancer agent, or 12 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours, 84 hours, or 96 hours, where any valuen can be a lower or upper endpoint of a range (e.g., 24 hours to 72 hours).

[0136] Disclosed methods comprise treating a subject with one or more other active ingredients, in addition to the pH-responsive nanoparticles described herein. For example, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with one or more additional drugs, such as a known anti-cancer drug. In an aspect, “coadministered” can be administration of a fixed dosage form combination drug comprising the pH-responsive nanoparticles described herein and a known anti-cancer drug. In an aspect, “coadministered” can be administration of separate dosage forms, e.g., the pH-responsive nanoparticles described herein as a solid oral dosage form and the anti-cancer drug as an intravenous dosage form. The pH-responsive nanoparticles described herein and other active agents can be administered separately or in conjunction with each other. In addition, the administration of the pH-responsive nanoparticles described herein can be prior to, concurrent to, or subsequent to the administration of another agent(s). The pH-responsive nanoparticles described herein and the other agent can be coadministered, either in concomitant therapy or in a fixed combination.

[0137] In an aspect, the pH-responsive nanoparticles described herein can be employed in combination with other known anticancer agents, and combinations thereof, and the like, or the pH-responsive nanoparticles described herein can be administered in conjunction with the use of physical methods such as with radiation therapy or surgery.

[0138] In an aspect, the pH-responsive nanoparticles described herein can be used as single agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of risk of the aforementioned diseases, disorders and conditions forATTORNEY DOCKET NO.930307-2000 which the pH-responsive nanoparticles described herein or the other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. The other drug(s) can be administered by a route and in an amount commonly used therefore, contemporaneously or sequentially with a disclosed compound. When the pH-responsive nanoparticles described herein is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the pH-responsive nanoparticles described herein is preferred. However, the combination therapy can also be administered on overlapping schedules. It is also envisioned that the combination of one or more active ingredients and the pH-responsive nanoparticles described herein can be more efficacious than either as a single agent.

[0139] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with a hormone therapy agent. In a still further aspect, the hormone therapy agent leuprolide, tamoxifen, raloxifene, megestrol, fulvestrant, triptorelin, medroxyprogesterone, letrozole, anastrozole, exemestane, bicalutamide, goserelin, histrelin, fluoxymesterone, estramustine, flutamide, toremifene, degarelix, nilutamide, abarelix, or testolactone, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0140] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with a chemotherapeutic agent. In a still further aspect, the chemotherapeutic agent coadministered in the disclosed methods is selected from one or more of the group consisting of an alkylating-like agent, an antimetabolite agent, an antineoplastic antibiotic agent, a mitotic inhibitor agent, an mTor inhibitor agent or another chemotherapeutic agent, or combinations thereof.

[0141] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with an antineoplastic antibiotic agent such as doxorubicin, mitoxantrone, bleomycin, daunorubicin, dactinomycin, epirubicin, idarubicin, plicamycin, mitomycin, pentostatin, or valrubicin, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0142] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with an antimetabolite agent such as gemcitabine, 5- fluorouracil, capecitabine, hydroxyurea, mercaptopurine, pemetrexed, fludarabine, nelarabine, cladribine, clofarabine, cytarabine, decitabine, pralatrexate, floxuridine, methotrexate, orATTORNEY DOCKET NO.930307-2000 thioguanine, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0143] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with an alkylating-like agent such as carboplatin, cisplatin, cyclophosphamide, chlorambucil, melphalan, carmustine, busulfan, lomustine, dacarbazine, oxaliplatin, ifosfamide, mechlorethamine, temozolomide, thiotepa, bendamustine, or streptozocin, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0144] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with a mitotic inhibitor agent such as etopside, vincristine, ixabepilone, vinorelbine, vinblastine, or teniposide, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0145] In an aspect, the pH-responsive nanoparticles described herein can be coadministered in the disclosed methods with a mTor inhibitor agent such as everolimus, siroliumus, or temsirolimus, or a pharmaceutically acceptable salt thereof, or combinations thereof.

[0146] The above combinations include combinations of the pH-responsive nanoparticles described herein not only with one other active compound, but also with two or more other active compounds. Likewise, the pH-responsive nanoparticles described herein can be used in combination with other drugs that are used in the prevention, treatment, control, amelioration, or reduction of risk of the diseases or conditions for which the pH-responsive nanoparticles described herein are useful. Such other drugs can be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with the pH-responsive nanoparticles described herein. When the pH-responsive nanoparticles described herein is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the pH-responsive nanoparticles described herein is preferred. Accordingly, the pharmaceutical compositions include those that also contain one or more other active ingredients, in addition to the pH-responsive nanoparticles described herein.

[0147] The weight ratio of the pH-responsive nanoparticles described herein to the second active ingredient can be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when the pH-responsive nanoparticles described herein is combined with another agent, the weight ratio of the pH- responsive nanoparticles described herein to the other agent will generally range from aboutATTORNEY DOCKET NO.930307-2000 1000:1 to about 1;1000, preferably about 200:1 to about 1:200. Combinations of the pH- responsive nanoparticles described herein and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used.

[0148] In the treatment of conditions such as cancer, an appropriate dosage level of astemizole in the pH-responsive nanoparticles described herein can be about 0.01 to 500 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level can be about 0.1 to about 250 mg / kg per day; more preferably about 0.5 to about 100 mg / kg per day. A suitable dosage level can be about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5 or 5 to 50 mg / kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10, 15.20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, and 1000 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The the pH-responsive nanoparticles described herein can be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosage regimen can be adjusted to provide the optimal therapeutic response. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient can be varied and will depend upon a variety of factors including the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

[0149] In an aspect of a disclosed method, the subject has been diagnosed with a need for treatment prior to the administering step. In an aspect of a disclosed method, the subject has been diagnosed with a disorder treatable by modulation of ANGPTL4 activity and / or a need for modulation of ANGPTL4 activity prior to the administering step. In an aspect of a disclosed method, the subject has been diagnosed with a disorder of uncontrolled cellular proliferation prior to the administering step. In an aspect, a subject can be treated prophylactically with the pH- responsive nanoparticles described herein.

[0150] Also provided are the uses of disclosed compounds and products. In an aspect, the disclosure pertains to use of at least one disclosed compound; or a pharmaceutically acceptable salt thereof. In an aspect, the compound used is a product of a disclosed method of making.ATTORNEY DOCKET NO.930307-2000

[0151] In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 50 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 25 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 10 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 5 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 500 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 250 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 100 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 50 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of less than about 10 nM.

[0152] In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of between from about 1 nM to about 50 µM. In an aspect, the pH- responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of between from about 1 nM to about 25 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of between from about 1 nM to about 10 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of between from about 1 nM to about 5 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit modulation of ANGPTL4 activity with an IC50of between from about 1 nM to about 1 µM.

[0153] In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 50 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 25 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 10 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 5 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 500 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 250 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibitionATTORNEY DOCKET NO.930307-2000 of cellular proliferation with an IC50of less than about 100 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 50 nM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of less than about 10 nM.

[0154] In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of between from about 1 nM to about 50 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of between from about 1 nM to about 25 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of between from about 1 nM to about 10 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of between from about 1 nM to about 5 µM. In an aspect, the pH-responsive nanoparticles described herein exhibit inhibition of cellular proliferation with an IC50of between from about 1 nM to about 1 µM. Kits

[0155] In an aspect, the disclosure pertains to kits comprising the pH-responsive nanoparticles described herein, and at least one of: (a) at least one agent known to increase ANGPTL4 activity; (b) at least one agent known to decrease ANGPTL4 activity; (c) at least one agent known to treat a disorder associated with ANGPTL4 activity; (d) instructions for treating a disorder associated with ANGPTL4 activity; (e) instructions for treating a disorder associated with ANGPTL4 activity; or (f) instructions for administering the compound in connection with treating a disorder of uncontrolled cellular proliferation.

[0156] In an aspect, the disclosure pertains to kits comprising at least one disclosed compound and at least one agent known to inhibit cellular proliferation.

[0157] In an aspect, the pH-responsive nanoparticles described herein and the at least one agent are co-formulated. In an aspect, the at least one compound and the at least one agent are co-packaged.

[0158] The kits can also comprise compounds and / or products co-packaged, co-formulated, and / or co-delivered with other components. For example, a drug manufacturer, a drug reseller, a physician, a compounding shop, or a pharmacist can provide a kit comprising a disclosed compound and / or product and another component for delivery to a patient.ATTORNEY DOCKET NO.930307-2000

[0159] In an aspect, the at least one agent is a hormone therapy agent. In an aspect, the hormone therapy agent is selected from one or more of the group consisting of leuprolide, tamoxifen, raloxifene, megestrol, fulvestrant, triptorelin, medroxyprogesterone, letrozole, anastrozole, exemestane, bicalutamide, goserelin, histrelin, fluoxymesterone, estramustine, flutamide, toremifene, degarelix, nilutamide, abarelix, and testolactone, or a pharmaceutically acceptable salt thereof.

[0160] In an aspect, the at least one agent is a chemotherapeutic agent. In an aspect, the chemotherapeutic agent is selected from one or more of the group consisting of an alkylating-like agent, an antimetabolite agent, an antineoplastic antibiotic agent, a mitotic inhibitor agent, an mTor inhibitor agent or other chemotherapeutic agent.

[0161] In an aspect, the antineoplastic antibiotic agent is selected from one or more of the group consisting of doxorubicin, mitoxantrone, bleomycin, daunorubicin, dactinomycin, epirubicin, idarubicin, plicamycin, mitomycin, pentostatin, and valrubicin, or a pharmaceutically acceptable salt thereof.

[0162] In an aspect, the antimetabolite agent is selected from one or more of the group consisting of gemcitabine, 5-fluorouracil, capecitabine, hydroxyurea, mercaptopurine, pemetrexed, fludarabine, nelarabine, cladribine, clofarabine, cytarabine, decitabine, pralatrexate, floxuridine, methotrexate, and thioguanine, or a pharmaceutically acceptable salt thereof.

[0163] In an aspect, the alkylating-like agent is selected from one or more of the group consisting of carboplatin, cisplatin, cyclophosphamide, chlorambucil, melphalan, carmustine, busulfan, lomustine, dacarbazine, oxaliplatin, ifosfamide, mechlorethamine, temozolomide, thiotepa, bendamustine, and streptozocin, or a pharmaceutically acceptable salt thereof.

[0164] In an aspect, the mitotic inhibitor agent is selected from one or more of the group consisting of etopside, vincristine, ixabepilone, vinorelbine, vinblastine, and teniposide, or a pharmaceutically acceptable salt thereof.

[0165] In an aspect, the mTor inhibitor agent is selected from one or more of the group consisting of everolimus, siroliumus, and temsirolimus, or a pharmaceutically acceptable salt thereof.

[0166] In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with surgery. In an aspect, the kit further comprisesATTORNEY DOCKET NO.930307-2000 instructions to provide the pH-responsive nanoparticles described herein in connection with surgery, wherein the instructions provide that surgery is performed prior to the administering the pH-responsive nanoparticles described herein. In an aspect, the kit further comprises instructions to provide the the pH-responsive nanoparticles described herein in connection with surgery, wherein the instructions provide that surgery is performed after the administering the pH- responsive nanoparticles described herein. In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with surgery, wherein the instructions provide that surgery is performed after the administering the pH-responsive nanoparticles described herein, and wherein the instructions provide that the administering the pH-responsive nanoparticles described herein is to effect presurgical debulking of a tumor. In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with surgery, wherein the instructions provide that surgery is performed after the administering the pH-responsive nanoparticles described herein, and wherein the instructions provide that surgery is performed at about the same time as the administering the pH-responsive nanoparticles described herein.

[0167] In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with radiotherapy. In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with radiotherapy, wherein the instructions provide that radiotherapy is performed prior to the administering of the pH-responsive nanoparticles described herein. In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with radiotherapy, wherein the instructions provide that radiotherapy is performed after the step of the administering the pH-responsive nanoparticles described herein. In an aspect, the kit further comprises instructions to provide the pH-responsive nanoparticles described herein in connection with radiotherapy, wherein the instructions provide that radiotherapy is performed at about the same time as the step of the administering the pH-responsive nanoparticles described herein.

[0168] In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent. In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent, and wherein each dose of the pH-ATTORNEY DOCKET NO.930307-2000 responsive nanoparticles described herein and the at least one agent are co-formulated. In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent, and wherein each dose of the the pH- responsive nanoparticles described herein and the at least one agent are co-packaged.

[0169] In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent, and wherein the dosage forms are formulated for oral administration and / or intravenous administration. In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the the pH-responsive nanoparticles described herein and the at least one agent, and wherein the dosage forms are formulated for oral administration. In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent, and wherein the dosage forms are formulated for intravenous administration.

[0170] In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent; and wherein the dosage form for the compound is formulated for oral administration and the dosage form for the at least one agent is formulated for intravenous administration. In an aspect, the kit further comprises a plurality of dosage forms, the plurality comprising one or more doses; wherein each dose comprises a therapeutically effective amount of the pH-responsive nanoparticles described herein and the at least one agent; and wherein the dosage form for the the pH-responsive nanoparticles described herein is formulated for intravenous administration and the dosage form for the at least one agent is formulated for oral administration.

[0171] In an aspect, the instructions for treating a disorder of uncontrolled cellular proliferation provide instructions for treating a cancer. It is understood that the disclosed kits can be prepared from the pH-responsive nanoparticles described herein, products, and pharmaceutical compositions. It is also understood that the disclosed kits can be employed in connection with the disclosed methods of using.ATTORNEY DOCKET NO.930307-2000 Aspects

[0172] Aspect 1. A composition comprising a pH-responsive nanoparticle having a core comprising astemizole or a pharmaceutically acceptable salt thereof encapsulated by a pH- responsive polymer carrier.

[0173] Aspect 2. The composition of Aspect 1, wherein the pH-responsive polymer carrier comprises a copolymer of poly(ethylene glycol) (PEG) and a poly(carbonate).

[0174] Aspect 3. The composition of Aspect 1, wherein the pH-responsive polymer carrier is a block copolymer of poly(ethylene glycol) (PEG) and the poly(carbonate).

[0175] Aspect 4. The composition of Aspect 1, wherein the pH-responsive polymer carrier has a structure represented by the formulawherein R1is C1-C4alkoxy, R2is C1-C12alkyl, C1-C12alkylamino, C2-C12dialkylamino, C3- C12cycloalkyl, or C2-C12heterocycloalkyl, and n and m are independently integers ranging from 10 to 300.

[0176] Aspect 5. The composition of Aspect 1, wherein the pH-responsive polymer carrier has one or mor structural units selected from the group consisting ofATTORNEY DOCKET NO.930307-2000 .polymer carrier is orinteger greater than 500 or a single integer greater than 1,000, or x is a single integer between 1 and 100, 1 and 1000, 1 and 10,000 or 1 and 1,000,000, or x is a single integer between 500 and a million, or x is a single integer between 100 and 100,000; andATTORNEY DOCKET NO.930307-2000 n is a single integer greater than 1, or n is a single integer greater than 500 or a single integer greater than 1,000, or n is a single integer between 1 and 100, 1 and 1000, 1 and 10,000 or 1 and 1,000,000, or n is a single integer between 500 and a million, or n is a single integer between 100 and 100,000.

[0178] Aspect 7. The composition of Aspect 1, wherein the pH-responsive polymer carrier comprises (a) a first polymer comprising a polysaccharide; and a second polymer comprising an alkylaminoalkyl ester or methacrylic acid grafted to the first polymer.

[0179] Aspect 8. The composition of Aspect 7, wherein the first polymer comprises a polysaccharide, wherein the polysaccharide has a degree of substitution between 0.5 and 3 per monomeric unit of the polysaccharide, wherein the first polymer is a polyhydroxyl polymer; the second polymer comprises a monomer selected from the group consisting of methacrylic acid and diethylaminoethyl methacrylic acid, wherein the second polymer is grafted to the first polymer; and a polyethoxylated sorbitan-based moiety covalently bound to the second polymer.

[0180] Aspect 9. The composition of Aspect 1, wherein the pH-responsive polymer carrier comprises a block copolymer comprising a hydrophilic polymer segment and a hydrophobic polymer segment, wherein the hydrophilic polymer segment comprises a polymer selected from the group consisting of: poly(ethylene oxide) (PEO), poly(methacrylate phosphatidyl choline) (MPC), and polyvinylpyrrolidone (PVP), wherein the hydrophilic polymer segment has from 100 to 130 monomer units, wherein the hydrophobic polymer segment comprises wherein R′ is H or CH3,ATTORNEY DOCKET NO.930307-2000 wherein R is NR1R2, wherein R1and R2are n-butyl, wherein n is 1 to about 10, and wherein x is about 20 to about 200 in total.

[0181] Aspect 10. A pharmaceutical composition comprising the pH-responsive nanoparticles of any one of Aspects 1-9 and a pharmaceutically acceptable carrier.

[0182] Aspect 11. A method for the treatment of a disorder of uncontrolled cellular proliferation in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of Aspects 1-9.

[0183] Aspect 12. The method of Aspect 11, wherein the mammal is a human.

[0184] Aspect 13. The method of Aspect 11, wherein the mammal has been diagnosed with a need for treatment of the disorder prior to the administering step.

[0185] Aspect 14. The method of Aspect 11, further comprising the step of identifying a mammal in need of treatment of the disorder.

[0186] Aspect 15. The method of Aspect 11, wherein the disorder is associated with ANGPTL4 expression.

[0187] Aspect 16. The method of Aspect 11, wherein the disorder is a cancer.

[0188] Aspect 17. The method of Aspect 16, wherein the cancer is a pancreatic cancer, breast cancer, renal cancer, gastric cancer, colorectal cancer, lymphoma, cancers of the brain, genitourinary tract cancer, lymphatic system cancer, stomach cancer, larynx cancer, lung, pancreatic cancer, and malignant melanoma.

[0189] Aspect 18. The method of Aspect 16, wherein the cancer is pancreatic cancer.

[0190] Aspect 19. The method of Aspect 18, wherein the pancreatic cancer is a gemcitabine- resistant pancreatic cancer.

[0191] Aspect 20. A method for inhibiting ANGPTL4 activity in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of Aspects 1-9.ATTORNEY DOCKET NO.930307-2000

[0192] Aspect 21. A method for inhibiting metastasis in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of Aspects 1-9.

[0193] Aspect 22. The method of any one of Aspects 11-21, wherein the mammal is further administered an anticancer agent.

[0194] Aspect 23. The method of Aspect 22, wherein the anticancer agent comprises a chemotherapeutic agent, an antineoplastic antibiotic agent, an antimetabolite agent, an alkylating agent, a mitotic inhibitor agent, or a mTor inhibitor agent.

[0195] Aspect 24. The method of Aspects 22 or 23, wherein the anticancer agent is gemcitabine.

[0196] Aspect 25. The method of any one of Aspects 22-24, wherein the pH-responsive nanoparticles are administered to the mammal prior to the administration of the anticancer agent.

[0197] Aspect 26. The method of any one of Aspects 22-24, wherein the pH-responsive nanoparticles are administered to mammal at least 12 hours prior to the administration of the anticancer agent.

[0198] Aspect 27. A kit comprising the pH-responsive nanoparticles of any one of Aspects 1- 9 and at least one of: (a) at least one agent known to increase ANGPTL4 activity; (b) at least one agent known to decrease ANGPTL4 activity; (c) at least one agent known to treat a disorder associated with ANGPTL4 activity; (d) instructions for treating a disorder associated with ANGPTL4 activity; (e) instructions for treating a disorder associated with ANGPTL4 activity (f) instructions for administering the compound in connection with treating a disorder of uncontrolled cellular proliferation.

[0199] Aspect 28. The kit of Aspect 27, wherein the at least one compound, the at least one product, or the at least one agent are co-formulated.

[0200] Aspect 29. The kit of Aspect 27, wherein the at least one compound, the at least oneATTORNEY DOCKET NO.930307-2000 product, or the at least one agent are co-packaged. EXAMPLES

[0201] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric.

[0202] PROPHETIC EXAMPLE 1 - Synthesis of pH-Responsive Block Polymers

[0203] PEG-bpoly(carbonate) is the major polymer precursor that was synthesized according to a previously described method using PEG (Mn = 5000 g / mol) as the macroinitiator. Briefly, ring- opening polymerization of penta fluorophenol-protected bis(methoxy propionic acid) derivative will be prepared according to Scheme 1 below. The resulting macromolecular precursor, PEG- bpoly(carbonate), will be post-functionalized stoichiometrically using N,N′-dibutylethylenediamine (pKa = 4.0) to yield the pH-responsive block copolymer PEG-DB (4) [PEG-b-poly- (carbonate) appended with N,N′-dibutylethylenediamine side chains.ATTORNEY DOCKET NO.930307-2000Scheme 1

[0204] PROPHETIC EXAMPLE 2 - Preparation of Astemizole-Loaded Nanoparticles from the pH-Responsive Block Copolymer

[0205] The self-assembled structures of amphiphilic block copolymers PEG-DB in the form of nanoparticles bearing astemizole will be prepared by a nonsolvent-induced phase separation (nanoprecipitation) method using DMSO as the selective solvent and PBS (pH 7.4) as the nonselective solvent. To form nanoparticles, 10 mg of PEG-DB and astemizole will be dissolved in 250 μL of DMSO. The resulting solution will be added dropwise to 750 μL of PBS buffer (pH 7.4), and the resultant solution will be transferred to a float-a-lyzer (MWCO = 3.5−5 kDa) to dialyze against ∼700 mL of PBS buffer (pH 7.4) overnight. The dialyzed suspension will be passed through a 0.2 μm PES filter to remove unencapsulated drugs to obtain PEG-DB nanoparticles (NPs). The particle size of the filtered, drug-loaded nanoparticle suspension will be measured using dynamic light scattering (DLS) studies at a scattering angle of 90°. The surface charge (ζ potential) of the carriers either in the presence or in the absence of 10% fetal bovine serum (FBS)ATTORNEY DOCKET NO.930307-2000 to determine the nanoparticles’ plasma stability as a function of time will also be measured.

[0206] It will be apparent to those skilled in the art that various modifications and variations can be made in the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.

Claims

ATTORNEY DOCKET NO.930307-2000 CLAIMS 1. A composition comprising a pH-responsive nanoparticle having a core comprising astemizole or a pharmaceutically acceptable salt thereof encapsulated by a pH- responsive polymer carrier.

2. The composition of claim 1, wherein the pH-responsive polymer carrier comprises a copolymer of poly(ethylene glycol) (PEG) and a poly(carbonate).

3. The composition of claim 1, wherein the pH-responsive polymer carrier comprises a block copolymer of poly(ethylene glycol) (PEG) and the poly(carbonate).

4. The composition of claim 1, wherein the pH-responsive polymer carrier has a structure represented by the formulawherein R1is C1-C4alkoxy, R2is C1-C12alkyl, C1-C12alkylamino, C2-C12dialkylamino, C3- C12cycloalkyl, or C2-C12heterocycloalkyl, and n and m are independently integers ranging from 10 to 300.

5. The composition of claim 1, wherein the pH-responsive polymer carrier has one or more structural units selected from the group consisting ofATTORNEY DOCKET NO.930307-2000 . 6.carrier is orn is a single integer between 100 and 100,000.ATTORNEY DOCKET NO.930307-2000 7. The composition of claim 1, wherein the pH-responsive polymer carrier comprises (a) a first polymer comprising a polysaccharide; and a second polymer comprising an alkylaminoalkyl ester or methacrylic acid grafted to the first polymer.

8. The composition of claim 7, wherein the first polymer comprises a polysaccharide, wherein the polysaccharide has a degree of substitution between 0.5 and 3 per monomeric unit of the polysaccharide, wherein the first polymer is a polyhydroxyl polymer; the second polymer comprises a monomer selected from the group consisting of methacrylic acid and diethylaminoethyl methacrylic acid, wherein the second polymer is grafted to the first polymer; and a polyethoxylated sorbitan-based moiety covalently bound to the second polymer.

9. The composition of claim 1, wherein the pH-responsive polymer carrier comprises a block copolymer comprising a hydrophilic polymer segment and a hydrophobic polymer segment, wherein the hydrophilic polymer segment comprises a polymer selected from the group consisting of: poly(ethylene oxide) (PEO), poly(methacrylate phosphatidyl choline) (MPC), and polyvinylpyrrolidone (PVP), wherein the hydrophilic polymer segment has from 100 to 130 monomer units, wherein the hydrophobic polymer segment comprises wherein R′ is H or CH3,wherein R is NR1R2, wherein R1and R2are n-butyl, wherein n is 1 to about 10, and wherein x is about 20 to about 200 in total.

10. A pharmaceutical composition comprising the pH-responsive nanoparticles of any one of claims 1-9 and a pharmaceutically acceptable carrier.ATTORNEY DOCKET NO.930307-2000 11. A method for the treatment of a disorder of uncontrolled cellular proliferation in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of claims 1-9.

12. The method of claim 11, wherein the mammal is a human.

13. The method of claim 11, wherein the mammal has been diagnosed with a need for treatment of the disorder prior to the administering step.

14. The method of claim 11, further comprising the step of identifying a mammal in need of treatment of the disorder.

15. The method of claim 11, wherein the disorder is associated with ANGPTL4 expression.

16. The method of claim 11, wherein the disorder is a cancer.

17. The method of claim 16, wherein the cancer is a pancreatic cancer, breast cancer, renal cancer, gastric cancer, colorectal cancer, lymphoma, cancers of the brain, genitourinary tract cancer, lymphatic system cancer, stomach cancer, larynx cancer, lung, pancreatic cancer, and malignant melanoma.

18. The method of claim 16, wherein the cancer is pancreatic cancer.

19. The method of claim 18, wherein the pancreatic cancer is a gemcitabine-resistant pancreatic cancer.

20. The method of any one of claims 11-19, wherein the mammal is further administered an anticancer agent.

21. The method of claim 20, wherein the anticancer agent comprises a chemotherapeutic agent, an antineoplastic antibiotic agent, an antimetabolite agent, an alkylating agent, a mitotic inhibitor agent, or a mTor inhibitor agent.

22. The method of claim 20, wherein the anticancer agent is gemcitabine.

23. The method of any one of claims 11-19, wherein the pH-responsive nanoparticles are administered to the mammal prior to the administration of the anticancer agent.

24. The method of any one of claims 11-19, wherein the pH-responsive nanoparticles are administered to mammal at least 12 hours prior to the administration of the anticancer agent.

25. A method for inhibiting ANGPTL4 activity in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of claims 1-9.

26. A method for inhibiting metastasis in a mammal comprising the step of administering to the mammal a therapeutically effective amount of the pH-responsive nanoparticles of any one of claims 1-9.ATTORNEY DOCKET NO.930307-2000 27. The method of claim 26, wherein the mammal is further administered an anticancer agent.

28. The method of claim 27, wherein the anticancer agent comprises a chemotherapeutic agent, an antineoplastic antibiotic agent, an antimetabolite agent, an alkylating agent, a mitotic inhibitor agent, or a mTor inhibitor agent.

29. The method of claim 27, wherein the anticancer agent is gemcitabine.

30. The method of claim 26, wherein the pH-responsive nanoparticles are administered to the mammal prior to the administration of the anticancer agent.

31. The method of claim 26, wherein the pH-responsive nanoparticles are administered to mammal at least 12 hours prior to the administration of the anticancer agent.

32. A kit comprising the pH-responsive nanoparticles of any one of claims 1-9 and at least one of: (a) at least one agent known to increase ANGPTL4 activity; (b) at least one agent known to decrease ANGPTL4 activity; (c) at least one agent known to treat a disorder associated with ANGPTL4 activity; (d) instructions for treating a disorder associated with ANGPTL4 activity; (e) instructions for treating a disorder associated with ANGPTL4 activity (f) instructions for administering the compound in connection with treating a disorder of uncontrolled cellular proliferation.

33. The kit of claim 32, wherein the at least one compound, the at least one product, or the at least one agent are co-formulated.

34. The kit of claim 32, wherein the at least one compound, the at least one product, or the at least one agent are co-packaged.

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