IL296531A - Method of treating glioma cancer using tarp gamma 8-dependent ampa receptor antagonists - Google Patents

Description

METHOD OF TREATING GLIOMA CANCER USING TARP GAMMA 8-DEPENDENT AMPA RECEPTOR ANTAGONISTS FIELD OF THE INVENTION id="p-1" id="p-1" id="p-1" id="p-1" id="p-1" id="p-1"

[0001] The embodiment ofs the present invention relat toe methods for treating a malignant glioma comprising administering a therapeutically-ef amoufectintve of a TARP yS-dependent AMPA receptor antagonist.

BACKGROUND OF THE INVENTION id="p-2" id="p-2" id="p-2" id="p-2" id="p-2" id="p-2"

[0002] Glial-derived tumors (/.e., gliomas) are a common type of tumo orir ginati inng the brain rath erthan a metastati brainc cancer. Abou t33% of all brain tumors are gliomas, which originate in the glial cell thats surround and suppo rtneurons in the brain includi, ngastrocytes, oligodendrocytes and ependymal cells Gli. omas are transformed cells that display increased metabol activitic asy a result of the transformatio process.n Neoplasti transformatic canon occur in all glia celll types, there byproducin a glarg rangee of patholog icaland morphological variants.

Most primary brain tumors derive fromd glia cellsl have lost growt controh regulal tion, giving rise to astrocytomas, glioblastomas, or oligodendrocyt omas.In the 2016 Worl dHealt hclassificat ion of brain tumors, there were no less than 36 different subtypes of glioma identified based, on histology and variou biomars kers.1 id="p-3" id="p-3" id="p-3" id="p-3" id="p-3" id="p-3"

[0003] Glioblastoma multifor (GBM)me , also known as glioblastoma, recurr entGMB, and/ograder IV astrocytoma, is an extremely aggressive tumo r.Symptoms are simila tor those of othe brainr tumors, and may include seizure, nausea and vomiting, headache, memory loss, hemiparesis, and progress memory,ive personality, or neurological defici duet to tempor aland fron tallobe involvemen Althot. ugh any brain tumo canr cause seizures, thei developmentr is most commonly associate withd neuroepithe tumorlial s,includi ngGBM. The incidence of epilepsy in patien tswith GBM varies betwee 30%n and 62%, in about two thirds as present ing symptom and in one third developing during the course of the disease.2 id="p-4" id="p-4" id="p-4" id="p-4" id="p-4" id="p-4"

[0004] It is very difficult to treat glioblastoma dues to several complicati factors.ng GBM tumo cellsr are generally very resistant to conventional therapies. These tumors conta inmany different type sof cells, where insome cells may respond well to certa thein rapies, whil eothers may not be affecte atd all .Another complicat ingfact oris that normal brai ncells are relativ ely susceptib tole damage due to conventional therapy, and the brain has a very limite dcapacity to repair itsel comparedf to other organs. The fact that many drugs cannot cross the blood-brain 1 barrier to act on the tumo furtherr limit sthe range of chemotherapie suits able for treating GBM patients. id="p-5" id="p-5" id="p-5" id="p-5" id="p-5" id="p-5"

[0005] GBM has the worst prognosi ofs any central nervous system (CNS) malignancy, despite multimodal treatity ment consisti ngof surgical resecti onof as much of the tumo asr possible, with concomitan or tsequential chemothera radiothpy, erapy, antiangiogenic therapy, immune therapy, gamma knife radiosurge andry, symptomat managemic ent wit h corticostero Prognoids. sisis very poor, with a median survival time of approximat oneely year and the disease is almost invariably fatal, as only about 3% survive for more than 3 years. id="p-6" id="p-6" id="p-6" id="p-6" id="p-6" id="p-6"

[0006] Accordingly, there is a need for new therapeu targtic ets and improved modaliti es for the treatmen of tmalignant gliomas, including GBM.

BRIEF SUMMARY OF THE INVENTION id="p-7" id="p-7" id="p-7" id="p-7" id="p-7" id="p-7"

[0007] The embodiments of the present invention provid ae method for treating a malignant glioma by administering an effective amount of a selective TARP y8 dependent AMPA recept antaor goni st.In addition to its beneficial anti-seizure/anti-epil therapeuticeptic effects, the administrat ofion a selective TARP y8-dependent AMPA recept antagor onist to a subject afflict withed a malignant glioma results in one or more of: (a) an alleviati ofon one or more symptom of the glioma; (b) a dela yor slowing of glioma tumo growtr and/oh metastasir s; (c) a stabilized state of a glioma tumo orr malignancy; and (d) an increased lifespan as compared to that expected in the absence of treatment. The malignant gliomas that can be treat edby the methods of the present invention include, but are not limite dto, astrocytoma , glioblastoma, oligodendrocytoma pilocytic, astrocytoma, diffuse intrinsi ponc tin glioma,e ependymom a,oligo-astrocytom oligodendrogliocytoma,a, optic pathway glioma, and hypothalamic glioma. id="p-8" id="p-8" id="p-8" id="p-8" id="p-8" id="p-8"

[0008] In one embodiment, the method for treating a malignant glioma in a subject comprise administs ering to the patient a pharmaceut icalcompositio comprin sing an effecti ve amount of Formul I:a HO—v Ch3 I 2 or a pharmaceutical acceptablly salte thereof. id="p-9" id="p-9" id="p-9" id="p-9" id="p-9" id="p-9"

[0009] In an alternate embodiment, the method for treating a malignant glioma in a subject comprises administering to the patien at pharmaceuti compositcal ion comprising an effective amount of Formula II: CH3 II or a pharmaceutical acceptablly salte thereof, where inX is CH or N; A is ; and R1 is select edfrom the grou consistip ngof: hydroge n, deuterium, fluoro, methyl, HO-(C1-C4 )-alky l,optionall substiy tute withd one or two methyl or deuterium groups, HO-(C1-C3)-alkoxy, optional substitly uted with one or two methyl or deuterium groups, fluoro-(C1-C3)-alkyl, HO-(C1-C3)-alkoxy-methyl, optionall substity uted wit hone or two methyl groups, cyano-(C1-C3)-alkoxy, HO-(C1-C3)-alkylthi optio, onall substity uted with one or two methyl groups, HO-(C1-C3)-alkyl-NH-, 3 HO-(C1-C3)-alkyl-N(CH3)-, methylsulfinyl , acyl, aminocarbonyl , methylcarbonylmethoxy methyl, aminomethylcarbonyloxyeth oxy, triazolylmethyl, 1 -methyl-imidizol-2-ylthio, -hydroxymethyl-tetrahydrofuran-2-yl, 3-hyd roxy-3-methylazet n-1 -ylidi, 3-methoxy-azetid -ylin-1 3-methoxy-3-methylazeti -yldin-1 4-hydroxypiperidin-1-yl, 4-hydroxy-4-methyl-piperidin-1 -yl, 4-hydroxy-4-vinyl-piperidin- 1-yl, 4-hydroxymethyl-piperidin- 1-yl, 4-(2-hydroxyethyl)-piperindi n-1-yl, morpholin-4-yl, 2-hydroxymethyl-morpholin-4 -yl, morpholin-4-yl-et hoxy,and tetrahydropyran- 4-yl, provide thatd when A is then R1 is not unsubstitute HO-(Cd 1-C3)-alkoxy, deuteriu substitm uted HO-(C1- C3)-alkoxy ,or HO-(C1-C3)-alkythio. id="p-10" id="p-10" id="p-10" id="p-10" id="p-10" id="p-10"

[0010] In alternat embodimentive thes, pharmaceut icalcompositio usedn in the treatment of a malignant glioma can further comprises one or more pharmaceutically acceptabl e carriers, diluents, or excipients. id="p-11" id="p-11" id="p-11" id="p-11" id="p-11" id="p-11"

[0011] In alternat embodimentive thes, metho dof the present invention is administered concomitantl or sequentialy withly one or more of a surgical resecti on,chemotherapy, 4 radiotherapy, antiangiogenic therapy, immune therapy, gamma knife radiosurgery, and symptomat managemic ent with corticosteroids. id="p-12" id="p-12" id="p-12" id="p-12" id="p-12" id="p-12"

[0012] Other implementatio arens also described and recit edherein.

BRIEF DESCRIPTION OF THE DRAWINGS id="p-13" id="p-13" id="p-13" id="p-13" id="p-13" id="p-13"

[0013] For the purpose of illustrat certaion, inembodiment ofs the present invention are shown in the drawing descris bed below. It shoul bed understood how, ever, that the invention is not limited to the precise arrangemen diments, sions, and instrument shown.s In the drawings: id="p-14" id="p-14" id="p-14" id="p-14" id="p-14" id="p-14"

[0014] FIG. 1 shows provides a diagrammat representaic tionof the 96-well plates showing the wells containi ngMedium (labeled "M"), the wells containi ngPBS (labeled "E"), and the fina concentratil mapon (pM) for the four test compounds: ES-481 and cisplati n concentrat areions shown in FIG. 1A; temozolom ideand perampan elconcentrati areons shown in FIG. 1B. id="p-15" id="p-15" id="p-15" id="p-15" id="p-15" id="p-15"

[0015] FIG. 2 shows the dose-response curves obtained in the LN-229 cell line with Temozolomide (...<....), Perampanel (——), ES-481 (... >:... ), and cisplat (in—▼—). id="p-16" id="p-16" id="p-16" id="p-16" id="p-16" id="p-16"

[0016] FIG. 3 shows the dose-response curves obtained in the U-87 MG cell line with Temozolomide (... ...... ), Perampanel (—B—), ES-481 (............ ), and cisplat (in—▼—). id="p-17" id="p-17" id="p-17" id="p-17" id="p-17" id="p-17"

[0017] FIG. 4 shows the dose-response curves obtained in the U-251 MG cell line with Temozolomide (...S... ), Perampanel (——), ES-481 ( ), and cisplat (in—▼—). id="p-18" id="p-18" id="p-18" id="p-18" id="p-18" id="p-18"

[0018] FIG. 5 shows the survival percenta curvesge obtained in the in vivo orthotopic U-87MG human glioblastoma xenograft mouse model with the vehicle (—•—) and 100 mg/kg Temozolomide given QD x 5/week for 2 weeks, p.o. (...•... ). id="p-19" id="p-19" id="p-19" id="p-19" id="p-19" id="p-19"

[0019] FIG. 6 shows the Kaplan-Meie lifr esurvival curves obtaine ind the in vivo orthotopic U-87MG human glioblastoma xenograft mouse model in mice treat edwith the vehicle (Group 1 - 20 ,(״ - ״ mg/kg ES-481 given QD x 5/week for 3 weeks, p.o. (Group 2------ ), ES-481 and 30 mg/kg Temozolomide given QD x 5/week for 2 weeks, p.o. combined with mg/kg ES-481 given QD x 5/week for 3 weeks, p.o. (Group 3.......).

DETAILED DESCRIPTION OF THE INVENTION id="p-20" id="p-20" id="p-20" id="p-20" id="p-20" id="p-20"

[0020] The subject innovation is now described with reference to the drawings, wherei n like reference numera lsare used to refer to like element thros ugho Inut. the followin g descripti on,for purposes of explanation numero, usspecifi detaic lsare set fort inh order to provid ae thorough understandi ofng the present invention. It may be evident howe, ver, that the present invention may be practiced without these specifi detac il s.In other instances, well - known structures and devices are shown in block diagram form in order to facilitat descrie bing the present invention. It is to be appreciated that certa inaspects, modes, embodiments, variations and features of the invention are described belo win various level ofs deta ilin order to provid ae substanti undeal rstandi ofng the present invention.

Definitions id="p-21" id="p-21" id="p-21" id="p-21" id="p-21" id="p-21"

[0021] For convenience, the meaning of some term sand phrases used in the specificati on,examples, and appended claims, are provided below. Unless state otherwise,d or implicit from context the, follow terming sand phrases include the meanings provided below.

The definitions are provide tod aid in describing particula embor diments and, are not intende tod limit the claime dinvention, because the scope of the invention is limite donly by the claims.

Unless otherwise defined, all technical and scientifi termc sused herein have the same meanin g as commonl undey rstood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definiti on provide herein,d the definition provide witd hin the specificat ionshall prevail. id="p-22" id="p-22" id="p-22" id="p-22" id="p-22" id="p-22"

[0022] As used in this specificat ionand the appended claims, the singul arform "a,"s "an" and "the" include plural referents unless the content clearl dicty ates otherwise. For example, reference to "a cell" includes a combination of two or more cells, and the like. id="p-23" id="p-23" id="p-23" id="p-23" id="p-23" id="p-23"

[0023] As used herein, the term "approximately or "about"" in referen toce a valu eor parameter are generall takeny to include numbers that fall within a range of 5%, 10%, 15%, or % in either direction (greater tha nor less than) of the number unless otherwise stated or otherwise evident from the context (except where such number would be less than 0% or exceed 100% of a possibl vale ue). As used herein, reference to "approximately or "about"" a valu eor parameter includes (and describes) embodiment thats are directed to that valu eor parameter. For example, descript ionreferring to "about X" includ esdescripti ofon "X". id="p-24" id="p-24" id="p-24" id="p-24" id="p-24" id="p-24"

[0024] As used herein, the term "or" means "and/or." The term "and/or as" used in a phrase such as "A and/or B" herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/o C"r is intended to encompass each of the following embodiment A,s: B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). id="p-25" id="p-25" id="p-25" id="p-25" id="p-25" id="p-25"

[0025] As used herein, the term "comprising" means that other element cans also be present in addition to the defined element presented.s The use of "comprising" indicates inclusion rathe thar nlimitation. 6 id="p-26" id="p-26" id="p-26" id="p-26" id="p-26" id="p-26"

[0026] The term "consisting of" refers to compositio ns,methods, and respective component theres asof described herein, which are exclusive of any element not recite ind that descripti ofon the embodiment. id="p-27" id="p-27" id="p-27" id="p-27" id="p-27" id="p-27"

[0027] As used herein the term "consisting essentially of" refers to those elemen ts required for a given embodiment. The term permits the presence of additional element thats do not materially affect the basic and novel or functional characteristic(s of that) embodiment of the invention. id="p-28" id="p-28" id="p-28" id="p-28" id="p-28" id="p-28"

[0028] The term "statisticall signify icant or "sig" nificantly" refers to statistical significance and generally means a two standard deviatio (2SDn ) or greater difference. id="p-29" id="p-29" id="p-29" id="p-29" id="p-29" id="p-29"

[0029] As used herein, the term "subject refers" to a mammal, includi ngbut not limited to a dog, cat, horse, cow, pig ,sheep, goat, chicken, rodent, or primate. Subjects can be house pets (e.g. ,dogs, cats), agricult uralstock animal s(e.g. ,cows, horses, pigs, chickens, etc.) , laborat oryanimal s(e.g. ,mice, rats, rabbit s,etc.) ,but are not so limited. Subjects include human subjects. The human subject may be a pediatric, adult or, a geriatri subject.c The human subject may be of either sex. The term s"subject" and "patient" are used interchangeabl herey in. id="p-30" id="p-30" id="p-30" id="p-30" id="p-30" id="p-30"

[0030] As used herein, the term s"effecti amount"ve and "therapeutically-eff ective amount" include an amount sufficient to prevent or ameliorate a manifestat ionof disease or medical condition, such as brain cancer. It wil lbe appreciat thated there wil lbe many ways known in the art to determine the effect iveamount for a given applicati on.For example, the pharmacologi methodscal for dosage determinat mayion be used in the therapeu conttic ext.

In the context of therapeu ortic prophylactic applications the, amount of a compositi on administered to the subject wil ldepend on the type and severit ofy the disease and on the characteristi of thecs individual such, as general health, age, sex, body weigh andt tolerance to drugs. It wil alsol depend on the degree, severity and type of disease. The skilled artisan will be able to determi neappropri atedosages depending on these and other factors. The compositions can also be administered in combination with one or more additional therapeutic compounds. id="p-31" id="p-31" id="p-31" id="p-31" id="p-31" id="p-31"

[0031] As used herein, the term s"treat" "treat, ment," "treating" or, "ameliorat" ionwhen used in reference to a disease, disorder or medical condition, refe tor therapeu treatmentstic for a condition, where inthe object is to reverse, alleviate ameli, orate, inhibit, slow down or stop the progression or severity of a symptom or condition. The term "treati"ng includes reducing or alleviatin at gleast one adverse effect or symptom of a condition. Treatment is generall y "effecti" veif one or more symptom ors clinica markersl are reduced. Alternatively treat, ment is 7 "effecti" veif the progression of a conditio isn reduced or halted. That is, "treatment" includes not just the improvement of symptom ors marker s,but also a cessati onor at least slowing of progress or worsening of symptoms that woul dbe expecte ind the absence of treatment .

Beneficia orl desire clind ical result includs e,but are not limite dto, alleviati ofon one or more symptom(s), diminishment of exten oft the deficit stabil, ized (/.e., not worsening) stat ofe a tumo r or malignancy, delay or slowing of tumor growt and/h or metastasis, and an increased lifespan as compared to that expected in the absence of treatment. id="p-32" id="p-32" id="p-32" id="p-32" id="p-32" id="p-32"

[0032] As used herein, the term "long-term" administrat meansion that the therapeuti c agent or drug is administered for a perio dof at least 12 weeks. This includ esthat the therapeu agenttic or drug is administere suchd that it is effective over, or for, a period of at least 12 weeks and does not necessari implyly that the administrat itselion takef splace for 12 weeks, e.g., if sustained release compositions or lon gacting therapeuti agentc or drug is used. Thus, the subject is treat edfor a period of at least 12 weeks. In many cases, long-ter admim nistrati on is for at least 4, 5, 6, 7, 8, 9 month ors more, or for at least 1,2, 3, 5, 7 or 10 years, or more. id="p-33" id="p-33" id="p-33" id="p-33" id="p-33" id="p-33"

[0033] The administrat ofion the compositions contemplat hereined may be carried out in any convenient manner, includi ngby aerosol inhalation injecti, on, ingestio transfn, usion, implantat ionor transplantatio In an. preferred embodiment, compositions are administered parenterall They. phrases "parenteral administrati" andon "administered parenterall" as usedy herein refers to modes of administrati othoner tha nenteral and topic aladministrat ionusuall, byy injectio n,and includes, without limitation, intravascu intravenous,lar, intramuscular, intraarteria l, intrathec intracapsulal, intraorbitalar, intratu, moral intracardiac,, intradermal, intraperitone al, transtracheal, subcutaneous subc, uticul intraar, articu subcapsullar, ar,subarachnoi intrd, aspin al and intrasternal injection and infusion. In one embodiment, the compositions contemplated herein are administered to a subjec byt direct injection into a tumor, lymph node, or site of infection. id="p-34" id="p-34" id="p-34" id="p-34" id="p-34" id="p-34"

[0034] The terms "decrease", "reduced", "reduction", or "inhibit" are all used herein to mean a decrease by a statistically significant amount In. some embodiments "reduce,, " "reduction" or "decrease" or "inhib"it typicall meansy a decrease by at least 10% as compared to a reference level (e.g., the absence of a given treatment or agent) and can includ e,for example, a decrea seby at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least abou 65%,t at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% , or more. As used herein, "reduct"ion or "inhibition" does not encompass a 8 complete inhibition or reduction as compare tod a reference level ".Complete inhibition" is a 100% inhibition as compared to a reference leve l.A decrease can be preferab downly to a leve l accepted as within the range of normal for an individu alwitho uta given disorder. id="p-35" id="p-35" id="p-35" id="p-35" id="p-35" id="p-35"

[0035] The terms "increased", "increase", "enhance", or "activat" aree all used herein to mean an increase by a statically significan amountt In. some embodiments the, terms "increased", "increase", "enhance", or "activat" cane mean an increase of at least 10% as compared to a reference level for, example an increase of at least about 20%, or at least about %, or at least about 40%, or at least abou 50%,t or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and includi nga 100% increase or any increase betwee n10-100% as compared to a reference leve l,or at least about a 2-fol d,or at least about a 3-fol d,or at least about a 4-fold, or at least about a 5-fol dor at least about a -fold increase, or any increase between 2-fol dand 10-fold or great eras compared to a reference level In. the context of a marker or symptom, a "increase" is a statistical signifly icant increase in such level. id="p-36" id="p-36" id="p-36" id="p-36" id="p-36" id="p-36"

[0036] As used herein, the term "cancer" relat esgenerally to a class of diseases or condition in swhich abnormal cells divide without contro andl can invad enearby tissues.

Cancer cell cans also sprea dto other part ofs the body through the bloo dand lymph systems.

There are several main type sof cancer Carci. noma is a cancer that begins in the skin or in tissues that line or cover internal organs. Sarcoma is a cance thatr begin sin bone, cartilage, fat, muscle, bloo dvessels, or other connect iveor support ivetissu e.Leukemia is a cancer that start ins blood-formi tingssue such as the bone marrow, and causes larg numberse of abnormal bloo dcells to be produced and enter the blood. Lymphoma and multipl myelomae are cancers that begin in the cells of the immune system. Central nervous system cancers are cancers that begin in the tissue ofs the brain and spinal cord. id="p-37" id="p-37" id="p-37" id="p-37" id="p-37" id="p-37"

[0037] In some embodiment ofs any of the aspects, the cance isr a primary cancer. In some embodiment ofs any of the aspect s,the cance isr a malignant cancer. As used herein, the term "malignan" referst to a cancer in which a grou ofp tumo cellr displays one or more of uncontrol growtled (/.e.h , division beyond normal limits) invasio, n(Le., intrusi onon and destruct ofion adjacent tissues), and metastasis (/.e., spread to other locations in the body via lymph or blood) As. used herein, the term "metastasi" zerefers to the spread of cance fromr one part of the body to another. A tumo formr ed by cell thats have spread is calle ad "metastat ic tumor" or a "metastasi" s.The metastat tumoic contr ains cells that are like those in the original (primary) tumor. 9 id="p-38" id="p-38" id="p-38" id="p-38" id="p-38" id="p-38"

[0038] As used herein, the term "benign" or "non-malignant" refers to tumors that may grow larger but do not sprea dto other part ofs the body. Benign tumors are self-limit anded typically do not invad eor metastasize. id="p-39" id="p-39" id="p-39" id="p-39" id="p-39" id="p-39"

[0039] A "cance cellr " or "tumo cellr " refers to an individu alcell of a cancero growtus orh tissu e.A tumo refersr generall to ya swelli ngor lesion formed by an abnorma growtl ofh cells, which may be benign, pre-malignan ort, malignan t.Most cance cellsr form tumors but, some, e.g., leukemia, do not necessari formly tumors For. those cancer cell thats form tumors the, term scancer (cell and) tumor (cell are) used interchangeably. id="p-40" id="p-40" id="p-40" id="p-40" id="p-40" id="p-40"

[0040] A subjec thatt has a cancer or a tumo isr a subjec havingt objectively measurable cance cellsr present in the subject’s body. Included in this definition are malignant, active ly proliferat cancers,ive as wel asl potentially dorma nttumors or micrometastatse Cancerss. which migrate from thei origir nal locati onand seed othe vitalr organs can eventual leadly to the death of the subject through the functional deteriorati ofon the affected organs. Hemopoieti c cancers, such as leukemia, are able to out-compet thee normal hemopoiet compartic ments in a subject, there byleading to hemopoiet failureic (in the form of anemia, thrombocytopenia and neutropeni ultia) matel causingy death. id="p-41" id="p-41" id="p-41" id="p-41" id="p-41" id="p-41"

[0041] Examples of cance inclr ude but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, leukemia, basal cell carcinoma, biliar tracty cancer; bladder cancer; bone cancer; brain and CNS cancer; breast cancer; cance ofr the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connect ivetissue cancer; cance ofr the digestive system; endometrial cancer; esophage cancer;al eye cancer; cance ofr the head and neck; gastri cancec (includir nggastrointesti cancer);nal glioblasto (GBM)ma ; hepati carcinoma;c hepatoma; intra-epithel neoplial asm; kidney or renal cancer; larynx cancer; leukemia; live r cancer; lun gcancer (e.g. ,small-cel lungl cancer, non-small cell lun gcancer, adenocarcino of ma the lung, and squamous carcinoma of the lung); lymphoma includi ngHodgkin’s and non­ Hodgkin’s lymphoma; melanoma; myeloma neurobl; astom orala; cavity cance (e.g.r ,lip, tongue, mouth, and pharynx); ovaria cancer;n pancreat cancer;ic prostate cancer; retinoblasto ma; rhabdomyosarcoma; rectal cancer; cancer of the respirat orysystem; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicula cancer;r thyro canceid r; uterin ore endometrial cancer; cancer of the urinary system; vulval cancer; as well as other carcinoma ands sarcomas; as wel asl B-cell lymphoma (includi nglow grade/follicul non­ar Hodgkin’s lymphoma (NHL); smal llymphocyt (SL)ic NHL; intermediat grade/e folli cularNHL; intermediat grade ediffuse NHL; high grade immunoblastic NHL; high grad elymphoblast NHicL; high grade smal lnon-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstro’s Macroglom buline chronicmia); lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chron myeloblastic leukeic mia; and post­ transplant lymphoproliferati disorderve (PTLD), as well as abnormal vascular proliferat ion associate withd phakomatoses, edema (such as that associate withd brain tumors and), Meigs’ syndrome. id="p-42" id="p-42" id="p-42" id="p-42" id="p-42" id="p-42"

[0042] A "cance cellr " is a cancerous, pre-cancerous, or transformed cell, either in vivo, ex vivo, or in tissue culture, that has spontaneous or induce dphenotypi changesc that do not necessari involvly thee uptake of new genetic material. Althoug transfh ormatio can arisen from infection wit ha transformin virusg and incorporat ofion new genomic nucleic acid, or uptake of exogenous nuclei acid,c it can also aris espontaneously or follow ingexposure to a carcinogen, there bymutating an endogenou gene.s Transformation/c isancer associate witd h, e.g., morphological changes, immortalizati ofon cells, aberra growtnt controlh foci, formati on, anchorage independence, malignancy, loss of contact inhibition and density limitation of growth, growt facth oror serum independence, tumor specifi markers,c invasiveness or metastasis, and tumo growtr inh suitab leanima lhost suchs as nude mice. id="p-43" id="p-43" id="p-43" id="p-43" id="p-43" id="p-43"

[0043] A subjec cant be one who has been previousl diagy nosed with or identifi edas sufferi fromng or havin ga condition in need of treatment (e.g. ,a cancer) or one or more complications relat edto such a condition, and optionall buty, need not have alread undergony e treatment for a condition or the one or more complicatio relatns edto the condition. Alternativ ely, a subjec cant also be one who has not been previously diagnosed as having a condition in need of treatment or one or more complications related to such a condition. For exampl e,a subject can be one who exhibit ones or more risk factors for a condition or one or more complications relat edto a condition or a subject who does not exhibit risk factors. A "subject in need" of treatment for a particular condition can be a subjec havingt that condition, diagnosed as having that condition, or at risk of developing that condition.

Pharmaceutical Compositions id="p-44" id="p-44" id="p-44" id="p-44" id="p-44" id="p-44"

[0044] The compositions and methods of the prese ntinvention may be utilized to treat an individual in need there of.In certa inembodiments the, individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceut icalcompositi on comprising, for example, a compound of the invention and a pharmaceutically acceptabl e carrier. Pharmaceutica acceptablelly carriers are well known in the art and include, for example, aqueous solutions such as water or physiological bufferedly saline or other solvents or vehicles such as glyco ls,glycerol, oils such as olive oil ,or injectab leorganic esters. In preferred 11 embodiments when, such pharmaceut icalcompositions are for human administrat ion, particularly for invasive routes of administratio (/.e.n, route suchs, as injection or implantation, that circumvent transport or diffusion through an epitheli barrier),al the aqueous soluti onis pyrogen-free, or substantiall pyrogy en-free. The excipient cans be chosen, for example, to effe ctdelayed release of an agent or to selectively targe onet or more cells, tissues or organs.

The pharmaceuti composical ti oncan be in dosage uni tform such as tablet, capsul (ine cludi ng sprinkle capsule and gelatin capsule), granule, lyophile for reconstitut powion,der, solution, syrup, suppository, injection or the like. The compositio cann also be present in a transderma l delivery system, e.g., a skin patch. The compositio cann also be present in a soluti onsuitabl e for topic aladministrati suchon, as a lotion cream,, or ointment. id="p-45" id="p-45" id="p-45" id="p-45" id="p-45" id="p-45"

[0045] A pharmaceutically acceptable carri ercan conta inphysiologically acceptabl e agents that act, for example, to stabilize increase, solubilit or yto increase the absorpti onof a compound such as a compound of the invention. Such physiologicall acceptabley agents include, for exampl e,carbohydrat suches, as glucose, sucrose or dextrans, antioxidant suchs, as ascorbic acid or glutathio chelatine, ngagents low, molecular weight proteins or other stabilizers or excipient s.The choice of a pharmaceutically acceptable carrier, including a physiologica acceptablelly agent, depends, for example, on the route of administrat ofion the composition. The preparati oron pharmaceut icalcompositio cann be a self-emulsif yingdrug delivery system or a self-micro emulsifyi ngdrug delivery system. The pharmaceuti cal composition (preparat ion)also can be a liposome or other polymer matri x,which can have incorpora thereited n,for example, a compound of the invention. Liposomes, for exampl e,which comprise phospholipi ords other lipids, are nontoxic, physiologica acceptablelly and metabolizabl carrierse that are relativel simply eto make and administer. id="p-46" id="p-46" id="p-46" id="p-46" id="p-46" id="p-46"

[0046] The phrase "pharmaceutically acceptabl ise" employed herein to refe tor thos e compounds, materials composi, tio ns,and/o dosager form whichs are, within the scope of sound medical judgment, suitable for use in conta ctwit hthe tissue ofs human being sand animals without excessive toxicit irrity, ation aller, gicresponse, or other problem or complication, commensurate with a reasonable benefit/risk ratio. id="p-47" id="p-47" id="p-47" id="p-47" id="p-47" id="p-47"

[0047] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, compositi onor vehicle, such as a liquid or soli dfiller, diluent, excipient solv, ent or encapsulating material. Each carri ermust be "acceptable" in the sense of being compatibl withe the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugar s,such as lactose, gluco seand sucrose; (2) starches, such as corn starc andh 12 potat starco h;(3) cellulose, and its derivatives, such as sodium carboxymet celluhyl lose, ethy l cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelati n;(7) talc; (8) excipient suchs, as cocoa butt erand suppository waxes; (9) oils, such as peanu oil,t cottonseed oil ,safflow oil,er sesame oil ,olive oil ,corn oil and soybean oil; (10) glyco ls,such as propylene glycol; (11) polyols, such as glyceri sorbitn, ol,mannitol and polyethylen glyce ol; (12) esters, such as ethyl oleate and ethyl laurat (13)e; agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) algini acid;c (16) pyrogen-f reewater; (17) isotoni salic ne; (18) Ringer' solutis on; (19) ethyl alcoho (20)l; phosphat buffee solutr ions; and (21) other non-toxic compatible substances employed in pharmaceut icalformulations. id="p-48" id="p-48" id="p-48" id="p-48" id="p-48" id="p-48"

[0048] A pharmaceut icalcompositi on(preparat ion)can be administered to a subjec byt any of a number of routes of administratio includin ng, for exampl e,oral ly(for example, drenches as in aqueous or non-aqueous solutio orns suspension tabls, ets, capsul es(includi ngsprinkle capsul esand gelat incapsules) boluses,, powders, granules, pastes for applicati onto the tongue); absorption through the oral mucosa (e.g., sublinguall subcuty); aneously; transdermally (for exampl eas a patc happlied to the skin); and topica lly(for example, as a cream, ointme ntor spray applied to the skin). The compound may also be formulat fored inhalation In .certai n embodiments a ,compound may be simply dissolved or suspended in steri lewater. Details of appropriat routese of administrat andion compositions suitable for same can be foun din, for example, U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231,5,427,798, 5,358,970 and 4,172,896, as well as in patent citeds therein. id="p-49" id="p-49" id="p-49" id="p-49" id="p-49" id="p-49"

[0049] The formulat ionsmay convenientl be ypresented in uni tdosage form and may be prepared by any methods wel knownl in the art of pharmacy. The amount of active ingredien t which can be combined with a carri ermaterial to produce a single dosage form wil varyl depending upon the host being treated, the particula moder of administrat ionThe. amount of active ingredient that can be combine dwit ha carrier materi alto produce a singl dosagee form wil general llybe that amount of the compound which produces a therapeuti effec ct. Generall y, out of one hundred percent, thi samount wil lrange from about 1 percent to about ninety-nin e percent of active ingredient, preferabl fromy about 5 percent to about 70 percent most, preferab fromly about 10 percent to abou 30t percent. id="p-50" id="p-50" id="p-50" id="p-50" id="p-50" id="p-50"

[0050] Methods of preparing these formulat ionsor compositions include the step of bringing into associati onan active compound, such as a compound of the invention, wit hthe carrier and, optional onely, or more accesso ingrry edients. In general, the formulat ionsare prepared by uniforml andy intimatel briny ging into associat iona compound of the presen t 13 invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product. id="p-51" id="p-51" id="p-51" id="p-51" id="p-51" id="p-51"

[0051] Formulations of the inventio suitabn lefor oral administrat mayion be in the form of capsul es(including sprinkl capsulese and gelat incapsules) cachet, s,pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-wat orer water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/o asr mouth washes and the like, each containing a predetermined amount of a compound of the prese ntinvention as an activ e ingredient. Compositio nsor compounds may also be administered as a bolus, electuary or paste. id="p-52" id="p-52" id="p-52" id="p-52" id="p-52" id="p-52"

[0052] To prepare solid dosage forms for oral administrat (capsuion les (includi ng sprinkle capsul esand gelat incapsules), tablets, pills, dragees, powders, granules and the like ), the active ingredient is mixed with one or more pharmaceutica acceptlly able carriers, such as sodium citrate or dicalcium phosphate, and/o anyr of the follow ing:(1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/o silr ici acid;c (2) binders, such as, for example, carboxymethylcellu alginlose,ates, gelati n,polyviny pyrrolidone,l sucrose and/or acacia; (3) humectants, such as glycero (4)l; disintegrat agents,ing such as agar-agar, calcium carbonate, potato or tapioca starc h,alginic acid, certa insilicates, and sodium carbonate; (5) solution retarding agents, such as paraffin (6); absorptio acceleratn ors,such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycer ol monostearat (8)e; absorbent suchs, as kaoli nand benton iteclay; (9) lubricant suchs, a talc , calcium stearate, magnesium stearate, solid polyethylene glyco ls,sodium lauryl sulfat ande, mixtur esthereof; (10) complexi ngagents, such as, modified and unmodifi edcyclodext rinsand ; (11) color ingagents. In the case of capsules (including sprinkle capsul esand gelati capsulen s), tablet ands pills, the pharmaceuti compositionscal may also comprise bufferi agentsng Solid. compositions of a similar type may also be employed as fillers in soft and hard-filled gelat in capsul esusing such excipient ass lactose or milk sugars, as well as high molecular weight polyethyl eneglycols and the like. id="p-53" id="p-53" id="p-53" id="p-53" id="p-53" id="p-53"

[0053] A tablet may be made by compression or molding, optional withly one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelat in or hydroxypropy methyll cellulose), lubrica nt,inert diluent, preservative, disintegrant (for example, sodium starch glycolat or ecross-linked sodium carboxymet cellulhyl ose), surfac­ e 14 active or dispersi ngagent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liqui ddiluent. id="p-54" id="p-54" id="p-54" id="p-54" id="p-54" id="p-54"

[0054] The tablets, and other soli ddosage forms of the pharmaceut icalcompositions, such as dragees, capsul es(includi ngsprinkle capsul esand gelat incapsule s),pills and granul es,may optionall bey scored or prepar edwith coatings and shell s,such as enteric coatings and other coatings well known in the pharmaceutical-formu art.lati Theyng may also be formulat soed as to provide slow or controlle relead se of the active ingredient therein using, for example, hydroxypropyl methyl cellulose in varying proportions to provid thee desired release profil othere, polyme matrir ces, liposom esand/o microspheres.r They may be sterilized by, for example, filtrati throon ugh a bacteria-retain filingter, or by incorporat steriing lizing agent s in the form of steri lesolid compositions that can be dissolved in steri lewater, or some other steri leinjectab lemedium immediately before use. These compositions may also optionall y conta inopacifyi ngagents and may be of a compositio thatn they release the active ingredient (s) only, or preferent iallin a y,certa inportion of the gastrointestina tract lopti, onal inly, a delayed manner. Examples of embedding compositions that can be used include polymeri substanc ces and waxes. The active ingredient can also be in micro-encapsulated form if, appropriate, wit h one or more of the above-described excipients. id="p-55" id="p-55" id="p-55" id="p-55" id="p-55" id="p-55"

[0055] Liquid dosage forms useful for oral administrat incluion de pharmaceutic ally acceptable emulsions, lyophiles for reconstitut microion, -emulsions solu, tio ns,suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form mays contai inertn diluents commonl usedy in the art such, as, for example, water or othe solvenr ts,cyclodextri ns and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcoh ol,isoprop yl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil s(in particul cottar, onse ed,groundn ut,corn, germ, olive, casto andr sesame oils), glycerol, tetra hydrofu alcohoryl polyetl, hylene glycols and fatty acid ester ofs sorbitan, and mixtures thereof. id="p-56" id="p-56" id="p-56" id="p-56" id="p-56" id="p-56"

[0056] Besides inert diluents, the oral compositions can also include adjuvant suchs as wetting agents, emulsifying and suspendin agents,g sweetenin flavog, ring, coloring, perfuming and preservati agents.ve id="p-57" id="p-57" id="p-57" id="p-57" id="p-57" id="p-57"

[0057] Suspensions, in addition to the active compounds, may conta insuspending agents as, for example, ethoxylat isosteed aryl alcohol polyos, xyethylene sorbit andol sorbit an esters, microcrystall celluine lose, aluminum metahydrox idebent, onit agar-e, agar and tragacanth and, mixtures thereof. id="p-58" id="p-58" id="p-58" id="p-58" id="p-58" id="p-58"

[0058] Dosage form fors the topical or transderm adminial strat incluion de powders, sprays, ointments, paste s,creams, lotions, gels, solutio ns,patches and inhalant s.The activ e compound may be mixed under steri leconditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers or ,propellant thats may be required. id="p-59" id="p-59" id="p-59" id="p-59" id="p-59" id="p-59"

[0059] The ointment pastes,s, creams and gels may contai n,in addition to an active compound, excipient suchs, as animal and vegetable fat s,oils, waxes, paraffin starch,s, tragacanth cell, ulose derivatives, polyethyle glycone ls,silicones, bentonites, silici acid,c talc and zinc oxide, or mixtures thereof. id="p-60" id="p-60" id="p-60" id="p-60" id="p-60" id="p-60"

[0060] Powders and sprays can contai n,in addition to an active compound, excipients such as lactose, talc sil, ici acid,c aluminu mhydroxide, calcium silicat esand polyamide powde r, or mixtures of these substances. Sprays can additional contaly incustomary propellan suchts, as chlorofluorohydrocarbons and volati unsule bstitute hydrocarbons,d such as butane and propane. id="p-61" id="p-61" id="p-61" id="p-61" id="p-61" id="p-61"

[0061] Transdermal patche haves the added advantage of providing controll delediver y of a compound of the prese ntinvention to the body. Such dosage form cans be made by dissolvi ngor dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controll byed either providing a rate control limembraneng or dispersi ngthe compound in a polyme matrixr or gel. id="p-62" id="p-62" id="p-62" id="p-62" id="p-62" id="p-62"

[0062] The phrases "parenteral administration and ""administered parenteral asly" used herein means modes of administrat othion er than enteral and topical administrat ionusuall, byy injectio n,and includes, without limitation, intravenous, intraocul (suchar as intravitre al), intramuscular, intraarteri intrathecalal, intracaps, ular, intraorbit intracardiac,al, intraderma l, intraperitonea transtl, racheal, subcutaneous, subcuticul intraar, articu subcapsular,lar, subarachnoi intd,raspina andl intrasternal injection and infusion Pharmaceut. icalcompositions suitable for parenteral administrat compriseion one or more active compounds in combination with one or more pharmaceutically acceptable steri leisotonic aqueous or nonaqueou solutis ons, dispersions, suspensions or emulsions, or steri lepowders which may be reconstituted int o steri leinjectab lesolutions or dispersions just prior to use, which may conta inantioxidants, buffers bacteriostats,, solut eswhich rende ther formulation isotoni withc the bloo dof the intended recipient or suspending or thickening agents. id="p-63" id="p-63" id="p-63" id="p-63" id="p-63" id="p-63"

[0063] Examples of suitable aqueous and nonaqueo uscarriers that may be employed in the pharmaceut icalcompositions of the invention include water, ethanol, polyol (suchs as glycerol, propylene glycol polyethyl, eneglycol, and the like), and suitab lemixtures thereof, 16 vegetable oils, such as olive oil ,and injectabl organice esters, such as ethyl oleat e.Examples of suitable aqueous and nonaqueou carris ers that may be employed in the pharmaceuti cal compositions of the invention include water etha, nol, polyol (suchs as glycerol, propylene glycol, polyethyl eneglycol, and the like), and suitable mixtures there of,vegetable oils, such as olive oil, and injectab leorgani esters,c such as ethyl oleat e.Proper fluidi canty be maintained for, example, by the use of coating material suchs, as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfacta nts.Proper fluidi tycan 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. id="p-64" id="p-64" id="p-64" id="p-64" id="p-64" id="p-64"

[0064] These compositions may also conta inadjuvant suchs as preservatives, wetting agents, emulsifyi ngagents and dispersin agents.g Prevention of the action of microorganisms may be ensured by the inclusion of variou ants ibacteri andal antifunga agents,l for example , paraben, chlorobutan phenolol, sorbic acid, and the like. It may also be desirabl toe include isotonic agents, such as sugars, sodium chloride and, the like into the compositio ns.In addition, prolonged absorptio ofn the injectable pharmaceut icalform may be broug htabout by the inclusion of agents that delay absorpti onsuch as aluminum monosteara andte gelatin. id="p-65" id="p-65" id="p-65" id="p-65" id="p-65" id="p-65"

[0065] In some cases, in order to prolong the effect of a drug, it is desirabl toe slow the absorpti onof the drug from subcutaneous or intramuscu injelarctio n.This may be accomplished by the use of a liqui dsuspension of crystalli orne amorphous material having poor water solubil ity.The rate of absorpt ionof the drug then depends upon its rate of dissolutio n, which in, turn, may depend upon cryst sizeal and crystal liform.ne Alternatively, delayed absorpti onof a parentera administlly ered drug form is accomplished by dissolvi ngor suspending the drug in an oil vehicle. id="p-66" id="p-66" id="p-66" id="p-66" id="p-66" id="p-66"

[0066] Injectabl depote forms are made by forming microencapsul atedmatric esof the subject compounds in biodegradable polymers such as polylactide-polygly Depencolide.ding on the rati oof drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be control led.Examples of othe biodegradabler polymers include poly(orthoester s) and poly(anhydrides). Depot injectable formulat ionsare also prepared by entrappi ngthe drug in liposomes or microemulsions that are compatib withle body tissue. id="p-67" id="p-67" id="p-67" id="p-67" id="p-67" id="p-67"

[0067] For use in the methods of thi sinvention, active compounds can be given perse or as a pharmaceuti compositcal ion containing, for example, 0.1 to 99.5% (more preferabl y, 0.5 to 90%) of active ingredie innt combination with a pharmaceutically-acceptabl carrier.e id="p-68" id="p-68" id="p-68" id="p-68" id="p-68" id="p-68"

[0068] Methods of introductio mayn also be provide byd rechargea orble biodegradabl e devices. Variou slows release polymeri devic ces have been develope andd tested in vivo in 17 recent years for the controlle delidvery of drugs, includi ngproteinaceou biospharmaceuti cals.

A variet ofy biocompati blepolymers (including hydrogels inclu), ding both biodegradable and non-degradab polymers,le can be used to form an implant for the sustained release of a compound at a particular target site. id="p-69" id="p-69" id="p-69" id="p-69" id="p-69" id="p-69"

[0069] Actual dosage levels of the active ingredients in the pharmaceuti compositcal ions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desire therapd euti responsec for a particular patient, composition, and mode of administrat ion, without being toxi cto the patient. id="p-70" id="p-70" id="p-70" id="p-70" id="p-70" id="p-70"

[0070] The select eddosage level wil dependl upon a variet ofy factors includi ngthe activi tyof the particula compoundr or combination of compounds employed, or the ester, salt or amide thereo thef, route of administrat ion,the time of administrat ionthe ,rate of excret ionof the particula compound(s)r being employed, the duration of the treatment, other drugs, compounds and/o materir als used in combination wit hthe particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts. id="p-71" id="p-71" id="p-71" id="p-71" id="p-71" id="p-71"

[0071] A physician or veterinaria havingn ordinar skilly in the art can readily determi ne and prescribe the therapeutical effelyctive amount of the pharmaceuti compositcal ion required.

For example, the physician or veterinaria couln dstart doses of the pharmaceut icalcompositi on or compound at level lows er than that required in order to achieve the desired therapeu effticect and gradually increase the dosage until the desired effe ctis achieved. By "therapeutically effective amount" is meant the concentrati ofon a compound that is sufficient to elicit the desired therapeu effetic ct. It is generally understoo thatd the effective amount of the compound wil varyl according to the weight, sex, age, and medica histol ryof the subject. Other factors which influence the effective amount may includ e,but are not limite dto, the severity of the patient's condition, the disorder being treate thed, stabili ofty the compound, and, if desire d,another type of therapeu agenttic being administered with the compound of the inventio n.A larger tota dosel can be delivered by multipl administratie onsof the agent. Methods to determi neefficacy and dosage are known to those skilled in the art .See, e.g., Isselbacher et al. (1996).3 id="p-72" id="p-72" id="p-72" id="p-72" id="p-72" id="p-72"

[0072] In general, a suitab ledaily dose of an active compound used in the compositions and methods of the invention wil lbe that amount of the compound that is the lowe stdose effective to produce a therapeu effticect. Such an effective dose wil generallyl depend upon the factors described above. id="p-73" id="p-73" id="p-73" id="p-73" id="p-73" id="p-73"

[0073] If desire d,the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separatel at yappropriat e 18 intervals througho theut day, optionall in y,unit dosage forms. In certa inembodiment ofs the present invention, the active compound may be administered two or thre tiemes daily. In other embodiments the, active compound will be administered once daily. id="p-74" id="p-74" id="p-74" id="p-74" id="p-74" id="p-74"

[0074] The patient receiving this treatment is any animal in need, includi ngprimates, in particula humansr and; other mammals such as equines bovine, porcine, sheep, feline and, canine; poultry; and pets in general. id="p-75" id="p-75" id="p-75" id="p-75" id="p-75" id="p-75"

[0075] In certa inembodiments compou, nds of the invention may be used alon eor conjointly administered with another type of therapeut agent.ic id="p-76" id="p-76" id="p-76" id="p-76" id="p-76" id="p-76"

[0076] The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certa inembodiments cont, emplate saltsd of the invention include, but are not limite dto, alkyl, dialkyl, trialkyl or tetra-al ammonikyl um salt s.In certa inembodiment conts, emplate saltd ofs the invention include, but are not limite dto, L-arginine, benenthamine, benzathin betaie, ne, calcium hydroxide, cholin e,deanol, diethanolamine, diethylami ne,2-(diethylamino)eth anol, ethanolami ne,ethylenediamin N-mete, hylglucamine, hydrabamine 1H-, imidazole, lithium, L-lysine, magnesium 4-(2-hydroxyethyl), morpholine piperazin, e,potassium, 1-(2- hydroxyethyl)pyrrolidi sodium,ne, triethanolamin trome, ethami ne,and zinc salt s.In certain embodiments contemp, lat saltsed of the invention include, but are not limite dto, Na, Ca, K, Mg, Zn or other metal salt s.In certa inembodiments, contemplat saltsed of the invention include, but are not limite dto, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoi acid,c 4-aminosalicylic acid, acet icacid, adipic acid , l-ascorbi acid,c I-asparti acid,c benzenesulfon acid,ic benzoic acid, (+)-camphori acidc , (+)-camphor-10-sulfonic acid, capric acid (decanoi acid),c capro icacid (hexanoic acid), capryli c acid (octanoi acid),c carbonic acid, cinnamic acid, citri acid,c cyclami acid,c dodecylsulf aciduric , ethane-1,2-disulfoni acid,c ethanesulf onicacid, formic acid, fumari acid,c galactari acidc , gentisic acid, d-glucohepton acid,ic d-gluconic acid, d-glucuronic acid, glutamic acid, glutar ic acid, glycerophospho acid,ric glycol acid,ic hippuri acid,c hydrobromi acid,c hydrochloric acid , isobutyri acid,c lacti acid,c lactobioni acid,c laur icacid, maleic acid, l-mal icacid, maloni acidc , mandeli acid,c methanesulfonic acid , naphthalen1,5-die- sulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitr icacid, oleic acid, oxali acid,c palmiti acid,c pamoic acid, phosphoric acid, proprioni acid,c l-pyrogluta acid,mic salicylic acid, sebaci cacid, stearic acid, succinic acid , sulfuri acid,c I-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroa acid,ceti andc undecylenic acid salts. 19 id="p-77" id="p-77" id="p-77" id="p-77" id="p-77" id="p-77"

[0077] The pharmaceutical acceply tabl acide addition salts can also exist as various solvat es,such as with water, methanol, ethanol, dimethylformam andide, the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallizati inherenton, in the solvent of preparati oron crystallizati or on,adventitious to such solvent. id="p-78" id="p-78" id="p-78" id="p-78" id="p-78" id="p-78"

[0078] Wetting agents, emulsifiers and lubricant suchs, as sodium lauryl sulfate and magnesium stearat ase, well as coloring agents rele, ase agents, coating agents sweet, ening, flavoring and perfuming agents preser, vati andves antioxidants can also be present in the compositions. id="p-79" id="p-79" id="p-79" id="p-79" id="p-79" id="p-79"

[0079] Examples of pharmaceuticall acceptabley antioxidan inclts ude: (1) water-solubl e antioxidan suchts, as ascorbi acid,c cystein hydrochlorie sodiumde, bisulfate sodium, metabisulfi sodite, um sulfi teand the like; (2) oil-soluble antioxidan suchts, as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylat hydroxytoled uene(BHT), lecithin, prop ylgallat alphe, ­a tocopherol, and the like; and (3) metal-chelatin agents,g such as citri acid,c ethylenediamine tetraaceti acidc (EDTA), sorbit ol,tartaric acid, phosphori acid,c and the like. id="p-80" id="p-80" id="p-80" id="p-80" id="p-80" id="p-80"

[0080] Unless otherwise defined herein, scientifi andc technical terms used in connection with the present applicat ionshall have the meanings that are commonl understy ood by those of ordinar skilly in the art to which thi sdisclosur beloe ngs. It shoul bed understood that thi sinvention is not limite dto the particular methodology, protoco andls, reagent etc.,s, described herein and as such can vary. The terminol ogyused herein is for the purpose of describi ngparticula embodimentr onlys and is not intended to limi tthe scope of the prese nt inventio whichn, is defined solel byy the claims. Definitions of common terms in immunology and molecular biology can be foun din The Merck Manual of Diagnosis and Therapy;4 The Encyclopedi ofa Molecular Cell Biolog andy Molecul Mediciar ne;5 Molecul Bioarlog andy Biotechnology: a Comprehensive Desk Reference;6 Immunolog7 y;Janeway's Immunobiolo8gy; Lewin's Genes XI;9 Molecul Cloning:ar A Laborato Manury al;10 Basic Methods in Molecul ar Biology;11 Laborato Methodsry in Enzymology;12 Current Protocols in Molecul Biolar ogy (CPMB);13 Curre ntProtocols in Prote inScience (CPPS);14 and Current Protocol in sImmunology (CPI).15 id="p-81" id="p-81" id="p-81" id="p-81" id="p-81" id="p-81"

[0081] Other term sare defined herein within the descripti ofon the various aspects of the invention.

Glutamate Receptors id="p-82" id="p-82" id="p-82" id="p-82" id="p-82" id="p-82"

[0082] Glutamate is the primar excity ator neurotransmity in mammaliter an brain.

Glutamatergi signalic ngparticipa tesin a wide range of neural functions includi nglearning and memory, long-te rmpotentiat andion synaptic plasticity. Glutamate receptors can be divided into two families The. ionotropic glutama receptorste form ion channels that activate upon binding agonist opening, a pore through the plasma membrane through which cations can flow. The metabotropic glutamate receptor ares G-protein-coup receptorled actis, vati ngintracellular signal transduct cascades.ion The ionotropic glutamate receptor cans be further subdivided into fou r sub-familie baseds, upon sequence homolo gyand selectivi toty exogenou agons ists. These sub-families are the AMPA a-amino-3-hydroxyl-5-methyl-4-isoxazole- acid),prop NMDionAic (N-methyl-D-aspart ate),kainate, and delta receptors. id="p-83" id="p-83" id="p-83" id="p-83" id="p-83" id="p-83"

[0083] The AMPA subtype of glutamat recepe tor ares glutama sensitite ve ion channels on postsynaptic membranes of excitatory synapses in the central nervous system (CNS) and are largely responsib forle mediatin fastg neurotransmissi acrosson synapt icgaps. AMPA receptor assembls ase tetramers of subunits. Mammals express four AMPA-receptor subunits, calle Glud A1-GluA 4.Each GluA subun itcan be expressed in multipl splicee variants; the two most prominent splice varian tsare called flop and flip .GluA subunits freel formy functional homo- and hetero-tetramers. The majorit ofy RNA encoding GluA2 subunits is edited post- transcription alterially, nga genetically-encoded glutamine to arginine. This RNA editing causes AMPA receptor to spreferenti formally wit htwo GluA2 units, and also prevents calciu mentr y through the activat edreceptor. id="p-84" id="p-84" id="p-84" id="p-84" id="p-84" id="p-84"

[0084] In thei natir ve environment, the pore-forming GluA tetrame direcrs tly or indirectl y associat withe numero usauxiliary protei nswhich modify the trafficking, localization, gating characteristics, and pharmacology of the AMPA recept (AMPAR)or . These auxiliary subunit s include cytoskelet andal anchori ngproteins, other signaling proteins, and several intracellul ar and transmembrane proteins with unknow functn ion. The wide variet ofy protei nswhich can participate in AMPA recept complexesor vastly increases the ability of a neuron to tune the response characteristi of itscs synapses. id="p-85" id="p-85" id="p-85" id="p-85" id="p-85" id="p-85"

[0085] Transmembrane AMPA Recept orRegulatory Proteins (TARPs) are a fairly recen tlydiscovered family of proteins that have been found to associate wit hand modula tethe activi tyof AMPA receptor Severals. TARPs are fairly regiospecifi in thec brain, leading to physiolog icaldifferentiatio of then AMPA recept activior ty. For example, TARP y2 (stargazi n) dependent AMPA receptor ares primar illocaly ized in the cerebellum and cerebral corte andx TARP y8 dependen AMPAt receptor ares localized primari inly the hippocampus, which region is 21 particularly relevant to seizure originati and/oon propagatr ion.It has been theorized that target ingindividu alTARPs may enable selective modulation of specifi brainc circu itswithout globa llyaffecti synaptng ictransmissio16 n.

AMPA Receptor Antagonists id="p-86" id="p-86" id="p-86" id="p-86" id="p-86" id="p-86"

[0086] AMPA recept antagonistsor are known anticonvulsant agents and thei abir lity to down modula teexcitator neuroty ransmissio is keyn to thei antir -epilepti therapc euti potc ential.

However, since AMPA recept activior tyis so ubiquito usin the CNS, general antagoni smaffec ts most areas of the CNS resulting in undesir edeffects, such as ataxia, sedation, and/or dizziness, which are shared by all known general AMPA receptor antagonists Typical. ly, these general antagonist haves very narrow therapeuti dosinc gwindows, meaning that typically the doses neede dto obtain anti-convulsant activit arey close to or overl apwith doses at which undesired effects are observed.17 id="p-87" id="p-87" id="p-87" id="p-87" id="p-87" id="p-87"

[0087] Levetiracetam ((S)-2-(2-oxopyrrolidin-1-yl)butanamide), gabapenti (2-[1-n (aminomethyl)cyclohexyl]acet acid), topiic ramate (2,3:4,5-Bis-O-(1-methylethylidene)-beta -D- fructopyranose sulfamate), and carbamazepine (5H-dibenzo[b,f]azepine-5-carboxami are de) current leading therapeu drugstic for epileptic seizures. None of the curren approvedtly drug s appear to act entirely through modulatio ofn AMPA receptors. id="p-88" id="p-88" id="p-88" id="p-88" id="p-88" id="p-88"

[0088] Talampan el((8R)-7-Acetyl-5-(4-aminophenyl)-8,9-dihydro-8-met,3-hyl-7H -1 dioxolo[4,5-h] [2,3]benzodiazepine), selurampanel (BGG492) (N-[7-isopropyl-6-(2-methyl-2H- pyrazol-3-yl)-2,4-dioxo-1,4-dihydro-2H-quinazolin-3-yl]methanesul and perafonampanmide el), (5'-(2-cyanophenyl)-1'-phenyl-2,3'-bipyridinyl-6'(TH)-one are general) (non-TARP dependent/non-selecti AMPAve) recepto antagr onists being tested as anti-epileptics. id="p-89" id="p-89" id="p-89" id="p-89" id="p-89" id="p-89"

[0089] Several selectiv none, -competitive TARP y8-dependent AMPA recept or antagonist weres subsequently developed to be effective anti-seizure/anti-epile therapeuticptic drugs without the side effects (e.g., sedation, ataxia, and/o dizzinr ess) of general (non-TARP dependent/non-selecti AMPAve) antagonists or TARP y2-dependent AMPA antagonists. Some selective, non-competi tiveTARP y8-dependent AMPA receptor antagonists are described in, for example, U.S. Patent Nos. 8,765,960 and 9,469,632. id="p-90" id="p-90" id="p-90" id="p-90" id="p-90" id="p-90"

[0090] AMPA recept antagonistsor are known anticonvulsant agents, and thei abilir ty to down-modulate excitatory neurotransmission is critical to thei antiepilr eptic therapeuti potentic al.

However, because AMPA recept activor ity is so ubiquito usin the central nervous system (CNS), a non-select AMPAive recept antaor goni approachst affects many areas of the CNS, resulting in undesired effects such as ataxia, sedation fal, ls, and/or dizziness, which are share dby all 22 known general or broad-spectrum AMPA recept antagonior sts. Typically, the doses of these medications needed to obtain anticonvuls activiant tyare close to, or overl apwith, doses at which undesired effects are observed. id="p-91" id="p-91" id="p-91" id="p-91" id="p-91" id="p-91"

[0091] TARPs are a fair lyrecentl discovey red family of proteins that have been foun dto associat withe and modulat thee activit ofy AMPA receptor TARP-s. y8-depende ntAMPA receptor ares localized primari inly the hippocampus a ,region of importance in complex partial seizures and particul arlrelevany tot seizur origie natio and/on propagation.r Research suggest s that selectively targeting individual TARPs may enable selecti vemodulation of specifi braic n circui witts hout global affly ecti ngsynaptic transmissio resuln, ting in improved efficacy, safety, and tolerability. id="p-92" id="p-92" id="p-92" id="p-92" id="p-92" id="p-92"

[0092] ES-481 (previousl knowny as CERC-611, LY3130481, LSN3130481,3130481) was discovered with the goal of retaini ngefficacy in epilept patientsic but avoiding the motori c adverse event (AEs)s that are a known liabili tyof global AMPA recept antaor gonists.

Important ES-481ly, is the first molecul knowne to selectively target and functional blockly region-spec AMPAific receptors after oral dosing. This selectivity was engineered into ES-481 by chemical structure-activit relatiyonshi (SAR)p studies to achieve selecti veblockade of AMPA receptor associates withd the auxiliary protei TARP-y8n (high densit iny hippocampus, a regio n of importance in partial epilepsies) whil esparin AMPAg receptor assocs iate witd hTARP-y2 (high density in cerebellum, which regulates the ataxi aand falli ngassociate witd hFycompa® (perampanel)). id="p-93" id="p-93" id="p-93" id="p-93" id="p-93" id="p-93"

[0093] ES-481 is the first molecule to selectively target TARP-y8-contain ingAMPA receptor Gis.ven the predominant hippocampal location of TARP-y8-dependent AMPA receptor thes, efficacy and side effect profile ES-481 may represent an improvement compared to current antiepilep drugstic and may potentiall treay higt h-grade gliomas..

Gliomas id="p-94" id="p-94" id="p-94" id="p-94" id="p-94" id="p-94"

[0094] Glial-derived tumors (/.e., gliomas) are a common type of tumo orir ginati inng the brain rath erthan a metastatic brain cancer. Abou t33% of all brain tumors are gliomas, which originate in the glial cell thats surround and suppo rtneurons in the brain, includi ngastrocytes, oligodendrocy andtes ependymal cells Gli. omas are transformed cells that display increas ed metabol activiic tyas a result of the transformat process.ion They comprise a diverse grou ofp neoplasms that diff erin thei morphology,r thei CNSr location, thei degr ree of invasiveness, their tenden cyfor progressio andn, thei growthr characteristi Neoplastcs. transfic ormatio can occurn in all glia celll types, thereby producin a glarg rangee of patholog icaland morphological variant s.

Most primary brain tumors derive fromd glial cell haves lost growt controh regulal tion, giving rise 23 to astrocytomas, glioblastomas, or oligodendrocytoma In thes. 2016 Worl dHealt hclassificat ion of brain tumors the, re were no less than 36 different subtypes of glioma identified based, on histology and various biomarkers.18 id="p-95" id="p-95" id="p-95" id="p-95" id="p-95" id="p-95"

[0095] Astrocytomas are glial cell tumors develope fromd connect ivetissue cells called astrocytes and are the most common primar intra-axialy brain tumo r,accountin forg nearl halfy of all primary brain tumors They. are most often foun din the cerebrum (the large, outer part of the brain) but, also in the cerebellum (locat edat the base of the brain). id="p-96" id="p-96" id="p-96" id="p-96" id="p-96" id="p-96"

[0096] Astrocytomas can develop in adult ors in children. High-grade astrocytomas, calle gliobd lasto mulma tiforme (GBM), are the most malignant of all brain tumors Gl. ioblastoma symptom ares often the same as those of other gliomas. Pilocytic astrocytomas are low-grade cerebell umgliomas commonl founy din children. In adults, astrocytomas are more common in the cerebrum. id="p-97" id="p-97" id="p-97" id="p-97" id="p-97" id="p-97"

[0097] Brain stem gliomas, also called diffuse infiltrati brainng stem gliomas or diffuse intrinsic pontin glie omas (DIPGs), are rare tumors found in the brain stem. They usually canno t be surgical removely becaused of thei remoter location, where they intertw inewit hnormal brai n tissue and affect the delicate and complex functio thins sarea controls. These tumors occur most often in school-age children where they are responsib forle the greatest number of childhood deaths from primary brain tumors. id="p-98" id="p-98" id="p-98" id="p-98" id="p-98" id="p-98"

[0098] Ependymomas develop from ependymal cells lining of the ventricles or in the spina cord.l Ependymomas are rare, accountin forg just 2% to 3% of primary brain tumors.

However, they account for about 8% to 10% of brain tumors in children and are more likely to affe ctthose younge thanr 10 years old. The most locati onfor ependymomas in children is near the cerebellum, where the tumo canr block the flow of the cerebral spina fluil dand cause increased pressure inside the skull (obstructive hydrocephal us).These tumors can sprea dto other parts of the brain or spina lcord (drop-metastases) due to the flo wof spina fluil d. id="p-99" id="p-99" id="p-99" id="p-99" id="p-99" id="p-99"

[0099] Mixed gliomas (also called oligo-astrocytomas) are made up of more than one type of glial cell Thei. r diagnosis as a distinct tumo typer is controversi andal may be resolved with genetic screening of tumo tisr sue. These tumors are often found in the cerebrum and are most common in adul men.t id="p-100" id="p-100" id="p-100" id="p-100" id="p-100" id="p-100"

[0100] Oligodendrogliomas form from oliogodendrocytes, the support ivetissue cell ofs the brain and are usually foun din the cerebrum. Abou t2% to 4% of primary brain tumors are oliogodendroglioma Theys. are most common in young and middle-ag edadults and more likely to occur in men. Seizures are a very common symptom of these gliomas (affecti 50%ng to 80% of patients) as, well as headache, weakness, or problems wit hspeech. Oligodendrogli omas 24 typically have a bette prognosisr than most other gliomas. They are significant morely favorable in term sof their biological behavior and thei prognosisr for patien tsand can have survivorshi p measured of decades for folks with these tumors that have the most favorabl biomarkere profiles. id="p-101" id="p-101" id="p-101" id="p-101" id="p-101" id="p-101"

[0101] Optic pathway gliomas are a type of low-grade tumo foundr in the optic nerve or chiasm. They often infiltrate the opti cnerves, which send messages from the eyes to the brain.

Peopl ewith neurofibromat areosis more likely to develop them. Optic nerve gliomas can cause vision loss and hormone problems, since these tumors are often locate atd the base of the brai n where hormonal contr isol located. Gliomas affecti nghormone functio mayn be known as hypothalamic gliomas. id="p-102" id="p-102" id="p-102" id="p-102" id="p-102" id="p-102"

[0102] The most importan determit nant of survival for gliomas is the "grade" of the glioma. Increasing grades represent increasing malignancy and decreasing differentiati on, which is associate witd hincreased mitoti activic tyand enhanced cell migration.19■20 Grade I tumors grow slowl andy can sometimes be totall removey byd surgery, while grad eIV tumors are fast-grow aggressiing, ve,and difficu tolt trea t.High-grade gliomas account for 30% of primary brain tumors in adults and are the second most common cause of cancer death in children under 15 years of age. High-grade gliomas are divided by grad einto two categori es: anaplast astrocic ytomas (WHO Grade III) and glioblasto mulma tiforme (GBM; WHO Grade IV).21 id="p-103" id="p-103" id="p-103" id="p-103" id="p-103" id="p-103"

[0103] GBM (Grade IV) tumors are characterize by thed presence of areas of necrotizi tissung thate are surround byed anaplast celic ls with numerous mitosi sand endotheli al proliferat Thision. characterist as ic,wel asl the presence of hyperplastic abnorma bloodl vessels, differenti atesthe tumo fromr Grade III astrocytoma whichs, do not have these features.

GBM tumors usually appea rin the cerebral whit ematter, grow quickly, and can become very larg befe ore causing symptoms. Less than 10% form more slowly, following dedifferentiation of low-grade astrocytoma or anaplast astrocytic oma. These are called secondary GBM tumors and are more common in younger patient (means age 45 versus 62 years). The tumo mayr extend into the meninges or ventricular wall, leading to high protei conten ntin the cerebrospin al flui d(CSF) (>100 mg/dL), as well as an occasional pleocytosis of 10 to 100 cells, mostly lymphocytes. Malignant cells carri edin the CSF may spread (rarely) to the spina lcord or cause meningeal gliomatosis. However, metastasi ofs GBM beyon dthe central nervous system is extremely unusual. Abou t50% of GBM tumors occupy more than one lobe of a hemisphere or are bilatera Tumorsl. of this type usually aris efrom the cerebrum and may rarel exhibity the classic infiltrati acrosson the corpus callosum, producing a butterf (billyateral gliom) a. id="p-104" id="p-104" id="p-104" id="p-104" id="p-104" id="p-104"

[0104] GBM has the worst prognosi ofs any central nervous system (CNS) malignancy, despite multimodal treatity ment consisti ngof surgical resecti onof as much of the tumo asr possible, with concomitan or tsequential chemothera radiothpy, erapy, antiangiogenic therapy, immune therapy, gamma knife radiosurge andry, symptomat managemic ent wit h corticosteroids. Progno sisis very poor, with a median survival time of approximatel oney year and the disease is almost invariably fatal, as only about 3% survive for more than 3 years.

Glioma -Associated Epilepsy id="p-105" id="p-105" id="p-105" id="p-105" id="p-105" id="p-105"

[0105] Glioma-associat epiled epsy is a type of symptomatic focal epilepsy, which may manifest with focal seizures, generalized seizures, or both .Epilepsy occurs in >80% of patients with a low-grade glioma and 40-60% of patient withs glioblastoma.22■23 It is often the fir stclear presentati ofon the glioma Anti. -neoplasti treatmentsc often cause amelioratio of nepilepsy, possibly by revert ingthe pathophysiolo gicalpro-epileptoge processesnic in the tumor. id="p-106" id="p-106" id="p-106" id="p-106" id="p-106" id="p-106"

[0106] Excessive glutamat releae se by glioma cells has been reporte tod induce pharmacologically-accessible neurona hyperel xcitati includion, ngepilepsy.24 Two recent reports by Venkataramani et al. (2019)25 and Venkatesh et al. (2019)26 suggest that neuron al hyperexcitat stimuion lat bonaes fide glutamater synapsgic es on glioma cells. Ionotropi c glutamat recepe tor actis vate intercellul calciumar signali ngnetworks to orchestrate glioma cell growt andh invasio n,presumably by facilitati oncogenicng signali ngcascades and cytoskel etal remodeling. id="p-107" id="p-107" id="p-107" id="p-107" id="p-107" id="p-107"

[0107] Research has recently start edto focu ons these pathophysiologi mechanismscal and on the possibiliti fores new mechanism-based anti-epilept treaic tmen ts.Two recen t investigations assessed the effects of perampanel (FYCOMPA®), the first antiepilep drugtic in the class of selecti venon-competiti antave goni ofst AMPA receptors,27 on glioma. Izumot oet al. (2019)28 assessed the effects of perampanel on the response to seizure and tumo­r progression in 13 patients wit hgliomas and uncontrollabl seizures.e Perampanel (4 mg/da yor 8 mg/day) was tested after surgery, radiation, and chemotherapy treatment Epils. epti seizuresc were reporte tod be eliminat edin eight of the 13 patients and volume reduction was detected during the 6 month period. Unfortunat theely, abstract does not specify if the radiatio andn chemotherapy overlapped wit hthe perampanel treatment. The authors concluded that perampanel treatment was effective at control litheng epilepsy and inhibiting tumor progression.

However, the abstract does not discuss the potentia contrl ibu tieffectsng of surgery, radiation, and chemotherapy treatment to sthe observed beneficial effects attributed to perampanel In. contrast to the alleg edbeneficial effects reported by Izumoto etal. (2019), a subsequent report by Mayer etal. (2020)29 indicate thatd perampanel inhibited epileptifor dischargesm in 26 organotypic brain slice ofs glioma but failed to attenuat tumoe growtr orh promote animal survival in rat C6 glioma model. id="p-108" id="p-108" id="p-108" id="p-108" id="p-108" id="p-108"

[0108] Even if perampanel was determined to be effective at attenuati tumorng growth and/o promr otin theg survival of subjects afflict withed a glioma, it shoul bed noted that this drug has significan advert se effects. The drug label has an FDA box warning that the drug cause may cause serious psychiat ricand behavioral changes; it may cause homicidal or suicidal thoughts.30 Other side effects have included dizziness, somnolence, vertigo, aggression, anger, loss of coordinat ionblur, red vision, irritabi litandy, slurred speech. Perampanel also reduces the effectivenes of levos norge streoral contraceptivesl by about 40%.31 Women who may get pregna ntshould not take it as studies in animals show it may harm a fetu s.Perampanel is liable to be abused; very high doses produced euphoria responses simila tor ketamine. As such, it is designate asd a Schedu leIII contro lledsubstance by the Drug Enforceme nt Administration32 Accordin. gly, these significan adverset effect wouls dlimi tthe therapeutic usefulness of perampanel for treating gliomas.

TARP y8-DEPENDENT AMPA RECEPTOR ANTAGONIST FOR THE TREATMENT OF GLIOMAS id="p-109" id="p-109" id="p-109" id="p-109" id="p-109" id="p-109"

[0109] Severe brain cance rshave been shown that integrat intoe the brain’s wiring.33■34 High-grade gliomas, includi ngglioblastoma, form synapses that hijack electr icalsignal froms healthy nerve cells to drive thei ownr growth. High-grade gliomas form synapses with healthy neurons that transmit electri calsignal tos the cancerous tissu e.The tumors also contai cell-to-n cell electri calconnections known as gap junctions. Togethe ther, two types of connections allow electrical signals from healthy nerve cells to be conducted into and amplified within the tumors. id="p-110" id="p-110" id="p-110" id="p-110" id="p-110" id="p-110"

[0110] High-grade gliomas include glioblastoma, a brain tumo seenr in adults that has a five-yea survivalr rate of 5%; diffuse intrinsi ponc tin glioe ma, a pediatric brain tumo withr a five- year survival rate belo w1%; and other diagnos essuch as pediatri glic oblastoma and diffus e midline gliomas occurring in the spina lcord and thalamus. Previous studies have indicated that high-gra degliomas use normal brain activit toy drive theigrowtr 35h.■36 id="p-111" id="p-111" id="p-111" id="p-111" id="p-111" id="p-111"

[0111] Existing drugs that block electr icalcurrent haves been foun dto reduce growt ofh high-gra degliomas. Perampanel, a seizur medicae tion that blocks activity of neurotransmit ter receptor on sthe receiving end of a synapse, was reported to reduce proliferation of pediatri c gliomas implant edinto mice by 50%. Meclofenamat a e,drug that blocks the action of gap junctions, result edin a simila decreaser in tumor proliferati37 on. id="p-112" id="p-112" id="p-112" id="p-112" id="p-112" id="p-112"

[0112] AMP-type glutamat recepte ors (AMPARs) are the principa transl duce ofrs fast synapt ictransmission in the mammalian CNS. When high-grade gliomas form synapses with 27 healthy neurons that transmit electr icalsignal tos the cancero tissues,us these connections allow electrical signals from healthy nerves to be conducted and amplified within the tumor; allowing tumo growth.r It was thus hypothesized that the use of selective TARP y8-dependent AMPA recept antagonor ists has the potent ialfor anti-tumorigenic effe ctby slowing down or inhibiting these electri calconnections. id="p-113" id="p-113" id="p-113" id="p-113" id="p-113" id="p-113"

[0113] The present invention provides that select iveTARP y8-dependent AMPA recept antor agonist cans be effective therapeu ticsfor the treatment of gliomas witho utthe undesired effects (e.g., sedation, ataxia, and/o dizzinr ess) of general (non-TARP dependent/non-selecti AMPAve) antagonists or TARP y2-dependent AMPA antagonists.

In addition to its beneficial anti-seizure/anti-epi therapeuleptic effticects, the administrat ofion a selecti veTARP y8-dependent AMPA recept antagonistor to a subject with a glioma wil lresult in one or more of: (a) an alleviatio of none or more symptom of the glioma; (b) a delay or slowing of glioma tumo growtr and/oh metar stasis; (c) a stabilized state of a glioma tumo orr malignancy; and (d) an increased lifespan as compared to that expecte ind the absence of treatment. id="p-114" id="p-114" id="p-114" id="p-114" id="p-114" id="p-114"

[0114] The present invention provides that TARP y8-depende ntAMPA recept or antagonist usefuls for the treatment of gliomas include a compound of Formul I:a HO—، ch3 I i.e., 6-((S)-1-{1-[5-(2-hydroxy-ethoxy)-pyridin-2-yl]-1/7-pyrazol-3-yl}-ethyl)-3/7-1,3-benzot hiazol- 2-one), or a pharmaceutically acceptable salt there of.This antagonist also, known as ES-481 (previousl knowny as CERC-611, LY3130481, LSN3130481, 3130481), is a potent and selecti veantagoni ofst transmembrane alpha-amino-3-hydroxy-5-met isoxazolehyl-4- prop ionic acid (AMPA) recept regulator oryprotei (n"TARP")-y8-dependent AMPA recept thator is being develope asd an adjunctive therapy for the treatmen of tpartial-onset seizure wits hor without secondarily generalized seizures in patien tswith epilepsy and for high-grade gliomas whose progression is robus tlreguly ated by neuronal activity. The present invention also provides the use of a pharmaceuti composical tio comprisingn a compound of Formul I,a or a 28 pharmaceutically-acceptable salt thereof and, one or more pharmaceutical acceptaly ble carriers, diluents, or excipients. id="p-115" id="p-115" id="p-115" id="p-115" id="p-115" id="p-115"

[0115] Additional TARP y8-dependent AMPA receptor antagonists useful for the method of the present invention for the treatment of gliomas include a compound of Formula II: CH3 II or a pharmaceutical acceptablly salte thereof, where inX is CH or N; A is ; and R1 is select edfrom the grou consistip ngof: hydroge n, deuterium, fluoro, methyl, HO-(C1-C4 )-alky l,optionall substiy tute withd one or two methyl or deuterium groups, HO-(C1-C3)-alkoxy, optional substitly uted with one or two methyl or deuterium groups, fluoro-(C1-C3)-alkyl, HO-(C1-C3)-alkoxy-methyl, optionall substity uted wit hone or two methyl groups, cyano-(C1-C3)-alkoxy, HO-(C1-C3)-alkylthi optio, onall substity uted with one or two methyl groups, HO-(C1-C3)-alkyl-NH-, 29 HO-(C1-C3)-alkyl-N(CH3)-, methylsulfinyl , acyl, aminocarbonyl , methylcarbonylmethoxy methyl, aminomethylcarbonyloxyeth oxy, triazolylmethyl, 1 -methyl-imidizol-2-ylthio, -hydroxymethyl-tetrahydrofuran-2-yl, 3-hyd roxy-3-methylazet n-1 -ylidi, 3-methoxy-azetid -ylin-1 3-methoxy-3-methylazeti -yldin-1 4-hydroxypiperidin-1-yl, 4-hydroxy-4-methyl-piperidin-1 -yl, 4-hydroxy-4-vinyl-piperidin- 1-yl, 4-hydroxymethyl-piperidin- 1-yl, 4-(2-hydroxyethyl)-piperindi n-1-yl, morpholin-4-yl, 2-hydroxymethyl-morpholin-4 -yl, morpholin-4-yl-et hoxy,and tetrahydropyran- 4-yl, provide thatd when A is then R1 is not unsubstitute HO-(Cd 1-C3)-alkoxy, deuteriu substitm uted HO-(C1- C3)-alkoxy, or HO-(C1-C3)-alkythio. id="p-116" id="p-116" id="p-116" id="p-116" id="p-116" id="p-116"

[0116] The present invention also provides the use of a pharmaceut icalcompositi on comprising a compound of Formula II, or a pharmaceutically acceptable salt there of,and one or more pharmaceutically-acceptable carriers, diluents, or excipients. id="p-117" id="p-117" id="p-117" id="p-117" id="p-117" id="p-117"

[0117] Accordi ngto the present invention, selecti veTARP yS-dependent AMPA recept or antagonist cans be used for the treatment of various gliomas including but not limite dto, astrocytoma, glioblastoma, oligodendrocytoma, pilocyti astrc ocytoma, diffuse intrinsi pontic ne glioma, ependymom a,oligo-astrocytom oligoda, endrogliocyt opticoma, pathway glioma, and hypothalamic glioma. id="p-118" id="p-118" id="p-118" id="p-118" id="p-118" id="p-118"

[0118] The main initial therapi esfor malignant gliomas and for glioblastoma typics ally start with a craniotomy and surgical resection for non-diffus glieomas involving pre-op and intra­ operative imaging, awake craniotomies or real-time monitoring of neurological condition. A tissue sample the ngoes to a pathologist and a name and histology is given for the tumo andr a grade .Important biomarkersly, are identified that not only help classif tumorsy but, also are predicti ofve response to certa therin apies. Treatment is tailor ed,as much as it can be, to the particula typer of glioma but most patients wil receivel external beam radiatio therapy.n Patients with many of the tumo typer swil lreceive the radiation combined wit hthe oral, alkylati agentng temozolomide, an oral alkylati agent.ng id="p-119" id="p-119" id="p-119" id="p-119" id="p-119" id="p-119"

[0119] Accordingly, the metho dof the present invention can include a combinati on therapy. The administrati ofon a selective TARP yS-dependent AMPA receptor antagonist for the treatment of a glioma can be done concomitant or lysequentially with one or more of a surgical resecti on,chemotherapy, radiotherapy antiangi, ogenic therapy, immune therap y, gamma knife radiosurgery, and symptomati managemc ent with corticosteroids. id="p-120" id="p-120" id="p-120" id="p-120" id="p-120" id="p-120"

[0120] Temozolomide (TMZ; bran dnames Temodar and Temodal and Temcad) is an oral chemotherapy drug. It is an alkylat ingagent widely used in the treatment of recurren t malignant gliomas. It is used as a second-l inetreatment for astrocytoma and as a first-l ine treatment for glioblasto multima forme. id="p-121" id="p-121" id="p-121" id="p-121" id="p-121" id="p-121"

[0121] The descript ionof embodiment ofs the disclosure is not intended to be exhaustive or to limi tthe disclosure to the precise form disclosed. While specifi embodic ment s of, and examples for, the disclosure are described herein for illustrati purpove ses, variou s equivalent modifications are possible withi nthe scope of the disclosure, as those skilled in the relevant art wil lrecogni ze.For example, while method steps or function ares presented in a given orde r,alternative embodiment mays perform function in sa different orde r,or functio ns may be perform substed anti allyconcurrent Thely. teachin gsof the disclosur providede herei n can be applied to other procedures or methods as appropria te.The various embodiment s described herein can be combined to provide further embodiments Aspec. ts of the disclosure can be modified, if necessary, to emplo ythe compositions, functions and concepts of the above references and applicat ionto provide yet further embodiment ofs the disclosure. Moreover due, to biologic functial onal equivalency consideratio somens, changes can be made in protein 31 structur withoute affecti theng biological or chemical action in kind or amount These. and other changes can be made to the disclosur in elight of the detailed descripti on.All such modifications are intended to be included within the scope of the appended claims. id="p-122" id="p-122" id="p-122" id="p-122" id="p-122" id="p-122"

[0122] Specific element ofs any of the foregoin embodimg ents can be combined or substituted for element ins other embodiment s.Furthermore, while advantages associat edwith certa inembodiment ofs the disclosu havere been described in the context of thes e embodiments othe, embodimentr mays also exhibit such advantages, and not all embodimen ts need necessari exhibly itsuch advantages to fall within the scope of the disclosure. id="p-123" id="p-123" id="p-123" id="p-123" id="p-123" id="p-123"

[0123] The technology describe hereind is further illustrat byed the follow ingexamples which in no way should be construed as being further limiting. Althoug methh ods and materia ls simila orr equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and material ares described below.

Human Exposure to ES-481 id="p-124" id="p-124" id="p-124" id="p-124" id="p-124" id="p-124"

[0124] ES-481 is current beinly g develope ford the treatment of seizure by the Applicant of the present invention. In the first Phase 1 study performed, a single ascending dose (3 mg, 6 mg, 12 mg, 25 mg, 37 mg, 50 mg, 62 mg, 75 mg, 87 mg, and 100 mg) is being administered in healthy adul volunteert in orders to asses sthe safet y,tolerabil andity, pharmacokinet ofics ES-481. To date, the single dose cohort ofs 3 mg, 6 mg, 12 mg, 25 mg, 37 mg, and 50 mg of ES-481 have completed dosin gin six active treatment (ES-481) and two placebo voluntee perrs cohort. All patients in the stud yare receiving continuous EEG monitoring prior to the administrat ofion the dose and up to 24 post-dos e.Only two mild adverse event weres possibly relat edto the administrat ofion ES-481: mild emesis was reported by one volunteer after receiving the 12 mg dose and dysgeusia (altered tast e)by one volunteer after receiving the 37 mg dose.

EXAMPLES id="p-125" id="p-125" id="p-125" id="p-125" id="p-125" id="p-125"

[0125] The invention now being generall described,y it wil lbe more readily understood by reference to the follow ingexamples which are included merely for purposes of illustration of certa inaspects and embodiment ofs the present invention and are not intended to limi tthe invention. 32 Example 1 Cell Viability Assessment of Three Cell Lines Following Treatment with ES-481 and Three Other Test Compounds: Temozolomide, Perampanel, and Cisplatin id="p-126" id="p-126" id="p-126" id="p-126" id="p-126" id="p-126"

[0126] The tumo growtr atth enuati poton ent ialof ES-481 and thre othe er test compounds, temozolomide, perampan el,and cisplat in,were assessed using a CellTiter-G lo® Luminescent Cell Viabilit Assayy (Promeg a)in thre celle lines: LN-229, U-87 MG, and U-251 MG. The CellTiter-Gl Lumino® escen Cellt Viabilit Assayy is a homogeneous method to determi nethe number of viable cell ins culture based on quantitat ofion the ATP present whi, ch signals the presence of metabolically active cells.

Materials Cell Lines id="p-127" id="p-127" id="p-127" id="p-127" id="p-127" id="p-127"

[0127] Three cell line swere used for the cell viability assessment LN-229,s: U-87 MG, and U-251 MG. These thre celle lines were derived from malignant gliomas and are commonly used for research on gliomas. id="p-128" id="p-128" id="p-128" id="p-128" id="p-128" id="p-128"

[0128] The LN-229 cell line (ATCC® CRL-2611 ™)38 is a human glioblasto cellma line that was established in 1979 from cells taken from a patient with right fronta paril eto-occipi tal glioblastoma. U-87 MG (ATCC® HTB-14™)39 is one of a number of cell lines derive fromd malignant gliomas by J. Ponte nand associate froms 1966 to 1969.40 U-251 MG cell line (Sigma product no. 09063001) is a human cell line derive fromd a malignant glioblastoma tumo byr explant technique. The U-251 cell line was established at the Wallenberg laborator Uppsaly, a, Sweden, more than 40 years ago and was derived from a male patien witht malignant astrocyto41ma. id="p-129" id="p-129" id="p-129" id="p-129" id="p-129" id="p-129"

[0129] The cell lines were cultured and, the cell viabili tywas assessed in 96-wells plate (Cat. No: 3610, Corning ).The culture medium used, and the seeding numbers per well are provide ind Table 1. All the cell line sare cultured at 37°C with 5% CO2.

Table 1 Medium Cell Name Seeding #/Well DMEM + 5% FBS 5000 LN-229 (MEM + 0.01 mM NEAA) + 10% FBS 7000 U-87 MG RPMI1640+ 10% FBS 3000 U-251 MG Reagents id="p-130" id="p-130" id="p-130" id="p-130" id="p-130" id="p-130"

[0130] The follow ingreagents were used in the cell viability assays: CellTiter-G (CTG)lo (Cat .No: G7573, Promega) 33 DMEM (Cat. No: SH30243.01, HyClone) RPMI1640 (Cat .No: SH30809.01, Hyclone) FBS (Cat. No: FND500, ExCell) DMSO (Cat. No: 0231-500ml, Amresco) NEAA (Cat. No: SH30238.01, Hyclone) MEM (Cat. No: SH30024.01) 0.25% Trypsin-EDTA (Gibco ,Cat. No: 25200-072, Lot. No: 2063861).

Equipment id="p-131" id="p-131" id="p-131" id="p-131" id="p-131" id="p-131"

[0131] The follow ingequipmen wast used in the cell viability assays: Vi-Cel lXR, Beckman Coult er(TACEL0030) Envision 2104 Multilabel Reader, PerkinElmer (USA) CO2 Incubato SANYOr, Reverse microscope, Chongguang XDS-1B, Chongqing Guangdian Corp..

Test Compounds id="p-132" id="p-132" id="p-132" id="p-132" id="p-132" id="p-132"

[0132] As mentioned above, ES-481 wil lbe compared to thre othere test compounds, temozolomide, perampan el,and cisplatin in a cell viability assessment assay. ES-481 is the selecti veTARP yS-dependent AMPA recept antaor goni st:6-((S)-1-{1-[5-(2-hydroxy-ethoxy)- pyridin-2-yl]-1/7-pyrazol-3-yl}-ethyl)-3/7-1,3-benzoth previouslyiazol-2-one characterize for dits anti-seizure/anti-epile effptiects.c42 id="p-133" id="p-133" id="p-133" id="p-133" id="p-133" id="p-133"

[0133] Temozolomide (TMZ; bran dnames Temodar and Temodal and Temcad) is an oral chemotherapy drug. It is an alkylat ingagent widely used in the treatment of recurren t malignant gliomas. It is used as a second-l inetreatment for astrocytoma and as a first-line treatment for glioblasto multima forme. id="p-134" id="p-134" id="p-134" id="p-134" id="p-134" id="p-134"

[0134] Perampanel (sold under the brand name FYCOMPA®) is an antiepileptic drug that is used in addition to other drugs to trea part tia seizul res and generalized tonic-clonic seizures for people older tha n12 years. It was the first antiepilepti drugc in the class of selecti venon-competiti antagonistve of AMPA receptor Ass. described above, its therapeutic potent ialto attenuate tumo growtr orh promote animal survival has recentl beeny investigat ined human and rats with mixed success. However, perampane’s well l-document signifed icant adverse effects limi tits potent ialusefulness in the treatment of glioma. id="p-135" id="p-135" id="p-135" id="p-135" id="p-135" id="p-135"

[0135] Cisplati wasn employed as a positive control for thi sstudy. Cisplatin is one of the most effective chemotherapeutic agents to date used for the treatment of many malignancies, includi ngglioma.43 Cisplati causesn tumor cell death by direct DNA damage and by generat ing 34 reactive oxygen intermediates. Cisplat inis the most common platinum chemotherapy drug; it was first approved in the U.S. in 1978. But cisplatin is not without its problem s.It has an extensive list of horrible side effects including severe nausea and vomitin g.These two side effects are, in fact so, bad that the drug's development was almost stopped when it was first tested on peopl e.In addition, tumors can become resistant to platinum-based chemotherapy drugs. id="p-136" id="p-136" id="p-136" id="p-136" id="p-136" id="p-136"

[0136] Relevant informati onon the four test compounds is provided in Table 2.

Table 2 MW Solvent Stock cone Work cone Compound Storage Temp 400 mM 1 mM RT 382.44 DMSO ES-481 100 mM 500 pM -20°C 194.15 DMSO Temozolomide 1 mM 349.38 DMSO 200 mM -20°C Perampanel 300.05 PBS 3.33 mM 3.33 mM 4°C Cisplatin Methods Day 1: Cell seeding id="p-137" id="p-137" id="p-137" id="p-137" id="p-137" id="p-137"

[0137] Each well of a 96-well plate contained 90 pL of cell suspension with standard medium. The number of cells to be seeded was determined on the basis of Cel lDensit y Optimizati onAssays.

Day 2: Drug treatment id="p-138" id="p-138" id="p-138" id="p-138" id="p-138" id="p-138"

[0138] Test compounds were serially-dilut (1:3)ed in 96-well plates at the concentrations indicated in FIG. 1. The test compounds (10 pL/well were) added to the corresponding plates (ES-481 and cisplat arein shown in FIG. 1A; temozolomide and perampanel are shown in FIG. 1B). The fina volul me was 100 pL/well for all the plates The. cell line swere cultured at 37°C with 5% CO2.

Day 5: Plate reading id="p-139" id="p-139" id="p-139" id="p-139" id="p-139" id="p-139"

[0139] After thre dayse of incubati onwith the test compounds, cell weres observed under a microsco tope ensure that cells treat edwith the vehicle contro werel in good condition.

Once confirmed, 50 pL of CellTiter-Gl Reao® gent was added to each well The. content weres mix for 2 minutes on an orbital shaker to facilitat celle lysis. The plat weree the nallowed to incubat ate room temperat urefor 10 minutes to stabilize luminescent signal ands the luminescence was recorded using an EnVision Mult Labeli Reader.

Data Analysis id="p-140" id="p-140" id="p-140" id="p-140" id="p-140" id="p-140"

[0140] Data analysis was performed using GraphPad Prism 7.0. In order to calculate the hal maxif mal effective concentrat (EC50)ion values, a dose-respo nsecurve was generate d using a nonlinea regressionr mode lwit ha sigmoidal dose response. The formul ofa survivi ng rate is shown below and, the EC50 values were automatica generally ted by GraphPad Prism 7.0.

/ LumTest article — LumM edium control \ The surviving rate (%) = ----- - ---------------- ;---- ------ —— -------------- ; x 100% \LumNone treated — LumMedium control/ Results id="p-141" id="p-141" id="p-141" id="p-141" id="p-141" id="p-141"

[0141] The dose response curves for ES-481, temozolomide, perampan el,and cisplati n in the cell lines LN-229, U-87 MG, and U-251 MG are shown in FIG. 2, FIG. 3, and FIG. 4, respectivel andy, the half maximal effective concentrati (EC50)on values are provide ind Table 3.

Table 3 EC50 (uM) Cell line ES-481 Perampanel Temozolomide Cisplatin LN229 89.99 174.59 NA 17.59 72.47 213.17 NA 6.32 U-87-MG 221.1 NA U251 117.33 5.45 id="p-142" id="p-142" id="p-142" id="p-142" id="p-142" id="p-142"

[0142] As expected, the most effect ivetest compound was the positive control, cisplatin, followed by ES-481, and the nperampanel and temozolomide. Althou ghcisplat displain yed the best tumo growtr attenh uati potenton ial(Le., the lowest EC50 values), this compound has well - known extensive list of horribl sidee effects. ES-481 was the second most effective test compound in all thre celle lines, ahead of perampanel and temozolomide. The least effecti ve test compound was temozolomide; EC50 values coul dnot be calculat ated the doses tested. id="p-143" id="p-143" id="p-143" id="p-143" id="p-143" id="p-143"

[0143] These data suggest that in, addition to its beneficial anti-seizure/anti-epileptic therapeu effticects, the administrat ofion a selecti veTARP yS-dependent AMPA recept or antagoni likest ES-481 woul dattenua tumote growtr inh subjects afflicted with a glioma thereb y producin oneg or more of the follow ingbenefici aleffects: (a) an alleviation of one or more symptom of the glioma; (b) a dela yor slowing of glioma tumor growt and/oh metastasir s; (c) a stabilized state of a glioma tumo orr malignancy; and/or (d) an increased lifespan as compared to that expected in the absence of treatment. 36 Example 2 In Vivo Efficacy Assessment of ES-481 and a combination of ES 481 and Temozolomide in the Orthotopic U-87 MG Human Glioblastoma Cancer Xenograft Model in Female BALB/c Nude Mice id="p-144" id="p-144" id="p-144" id="p-144" id="p-144" id="p-144"

[0144] The present stud yevaluat edpreclinically the in vivo therapeu effictic acy of ES-481 and a combination of ES-481 and temozolomi inde the orthotopic U-87 MG human glioblastoma cance xenogrr aftmodel in BALB/c nude mice.

Experimental Methods Animals id="p-145" id="p-145" id="p-145" id="p-145" id="p-145" id="p-145"

[0145] Female Mus musculus BALB/c nude mice (n=30) were obtained from Beijing Anikeeper Biotech Co., Ltd (Beijing, China). At the initiation of the study, mice were 7-8 weeks old (estimat edage at inoculati on)wit ha body weigh oft 17.0-21.9 g. Mice were housed in Polysulfon IVCe cage (325 mm * 210 mm * 180 mm) up to 4 mice per cage with cardboard cylinder tissu, epaper, and PC tube/club house for environment enrichment and crushed cornc obbedding, autoclaved changed; weekl y.The housing temperature was 20-26°C wit ha 40-70% humidit levely and a light cycle of 12 hour light (7:00 am-7:00 pm) and 12 hours dark.

Mice were provided 0.2 pm filtered reverse, osmosis autoclaved, water and fed standard rodent chow, irradiated, ad libitum.

Cell Culture id="p-146" id="p-146" id="p-146" id="p-146" id="p-146" id="p-146"

[0146] The U-87 MG tumo cellr weres maintained in vitro as a monolaye cultr ure in DMEM medium supplemented with 10% heat inactivat feted al bovin eserum at 37°C in an atmosphe ofre 5% CO2 in air. The cells, when in exponent ialgrowt phase,h were harvest edand quantitated by cell count erbefore tumo inoculatr ion.

Selection of Dose id="p-147" id="p-147" id="p-147" id="p-147" id="p-147" id="p-147"

[0147] The dose of ES-481 selected was based on the anti-seizure doses previousl y documented in a mice seizure model s.In addition, the dose of 20 mg/kg ES-481 was also chosen because the expecte antid -seizu dosere in humans is though tot range from approximat 80ely mg to 150 mg per day of ES-481 and thi sdose shoul bed sufficient to inhibit hyperactive electrical conduction in cancerous glioma cells To. assess the potent ialcontribution of ES-481 in combination with temozolom idein this orthotopi xenograftc model a, dose of mg/kg x 5/week x 2 week of temozolom idewas select edin combination with 20 mg/kg x /week x 3 weeks of ES-481. 37 Tumor Inoculation id="p-148" id="p-148" id="p-148" id="p-148" id="p-148" id="p-148"

[0148] After anesthesia by intraperitoneal injection of Ketamine (0.1mL/mouse), head skin of the mouse was sterilized with the 75% alcoh oland then a 2 to 3 mm length incision was made just at the right of the midline and anteri toor the interaur line.al Mice were inoculated intracranially with the tumo cellsr (2 x1q5 cells in 2 pL PBS) at the right fron tallobe 2 mm lateral from the bregma and 0.5 mm from the anteri orat a dept hof 3.5 mm. The incision was stitched using No.6 suture and then sterilize witd hpovidone iodine solution. The mice were kept warm unti theyl recovered from anesthesia The. treatments were start edat day 5 after randomizat ion.

Each grou consistedp of 10 mice.

Randomization id="p-149" id="p-149" id="p-149" id="p-149" id="p-149" id="p-149"

[0149] Before grouping and treatment, all animal swere weighed, body weigh wast used as numeric parameter to randomize select edanimals into specified groups. Thus, the systematic error was minimized. id="p-150" id="p-150" id="p-150" id="p-150" id="p-150" id="p-150"

[0150] The randomizati startedon at Day-5 after inoculat ionThi. rty (30) mice were enrolled in the study. All animal swere randomly allocate to dthre treate ment groups described in Table 4. Randomization is performed based on "Matched distribut" ionmethod/ "Stratifi" ed metho d(StudyDirecto softwr™ are, version 3.1.399.19).

Table 4 Dose Dosing Vol Dosing Frequency Group n Treatment ROA (mg/kg) (pL/g) & Duration Vehicle* 0 10 P.O. 5x/week, 3 weeks 1 ES 481 25 10 P.O. 5x/week, 3 weeks 2 Temozolomide 75 10 P.O. 5x/week, 3 weeks 3 10 ES 481 25 10 P.O. 5x/week, 3 weeks * Vehicle: 20% (w/v) HPBCD in 25 mM phosphate buffer (pH 9) ROA: Route of Administratio is oraln or per os (P.O.).

Observation and Data Collection id="p-151" id="p-151" id="p-151" id="p-151" id="p-151" id="p-151"

[0151] After tumo cellsr inoculatio then, animals were checked dail yfor morbidit andy mortali ty.During routin monie tori ng,the animal swere checked for any effects of tumo growr th and treatments on behavior such as mobility, food and water consumption body, weight gain/loss (body weights were measured every other day afte randomr ization), eye/hair matting and any othe abnormalir ti es.Mortalit andy observe clinid cal signs were recorded for individu al animals in detail. Body weights and tumo volumesr were measured using StudyDirect or™ software (version 3.1.399.19). 38 Experimental Termination Study Endpoints id="p-152" id="p-152" id="p-152" id="p-152" id="p-152" id="p-152"

[0152] The major endpoin wast animal survival. The survival of all animal swere monitored and the median survival time (MST) was calculat fored each grou p.The increase in life-span (ILS) was calculat ased follows: _ 1nn x [Median Survival Time of drug treat edgroup _ 10, נ/, Median Survival Time of vehicle grou p ' °' Study Termination id="p-153" id="p-153" id="p-153" id="p-153" id="p-153" id="p-153"

[0153] The observation time for the survival study was 40 days after inoculat ionThe. brain tumo wasr remove afterd euthanasia and weighed. The differences in tumo weir ghts among groups were analyzed for significance using the ANOVA test.

Body Weight Loss id="p-154" id="p-154" id="p-154" id="p-154" id="p-154" id="p-154"

[0154] The body weigh oft all animal swere monitore throughoutd the stud andy animals were euthanized if they lost over 20% of thei bodyr weight relative to the weight at the first day of treatment.

General Animal Welfare Surveillance id="p-155" id="p-155" id="p-155" id="p-155" id="p-155" id="p-155"

[0155] All animal swere monitored throughout the stud andy animal swere euthanized if any of the following signs are observed: severe dehydration hypothermia,, abnormal/labored respirati on,lethargy, obvious pain, diarrhea, skin lesions, neurological symptoms, impaired mobili ty(not able to eat or drink due) to significant ascites and enlarged abdomen asta, sia , continuous prone or lateral position, signs of muscular atrophy, paralyti gait,c cloni c convulsions, toni cconvulsions, persistent bleeding from body orifice.

Statistical Analysis id="p-156" id="p-156" id="p-156" id="p-156" id="p-156" id="p-156"

[0156] For survival analysis, Kaplan-Mei ersurvival curves were generat ed,and a Log Rank test was performed The. event of interest was the anima ldeath. The survival time was defined as the time from the day of tumo cellr inoculati untilon one day before animal death or ethical endpoint. For each grou p,the median survival time (MST), corresponding 95% confidence interval and the increased in life-span (ILS) were calculated. All data were analyzed using SPSS 18.0 and/or GraphPa dPrism 5.0. P < 0.05 was considered statisticall signiy ficant. 39 Results Historical Data with this Model id="p-157" id="p-157" id="p-157" id="p-157" id="p-157" id="p-157"

[0157] Prio tor initiati ngthe prese ntin vivo orthotopic U-87MG human glioblastoma xenograft mouse model study with ES-481, it was well documented in thi smodel tha t,with a temozolomide dose of 75 mg/kg (data not shown) and 100 mg/kg x 5days/week x 2 week (FIG. 5), no animal survived beyond Day 30.

ES 481 Alone or In Combination with Temozolomide id="p-158" id="p-158" id="p-158" id="p-158" id="p-158" id="p-158"

[0158] Althou ghthere was no statistically-signif differicantence in overa survill val betwee vehiclen (Group 1) and ES-481 (Group 2), there was a statistically-signif dificanferencet in the weigh oft the tumors betwee Groupn 1 and Group 2. More impressive was the fact that all of the animal sin ES-481 and temozolomi (Groupde 3) survived and had no tumors presented when sacrificed at Day 41. Table 5 provides the tumo weigr hts and FIG. 6 shows the overal l survival curves for the thre groue ps.

Table 5: Tumor Weights (mg) Group 1 Group 2 Group 3 Vehicle ES-481 ES-481 and Temozolomide Number of Animals 10 10 10 Missing Data 1 0 0 Mean SD 122.6(134.7) 58.8 (84.53) 0(0) Median 110.2 18.2 0 Min, Max 0, 357.1 0, 255 0,0 95% Cl 39.1, 206 6.5, 111 0,0 Abbreviations: SD = Standar Devid ation; Cl = Confidence Interval id="p-159" id="p-159" id="p-159" id="p-159" id="p-159" id="p-159"

[0159] These data suggest that in, addition to its beneficial anti-seizure/anti-epileptic therapeu effticects, the administrat ofion a selecti veTARP y8-dependent AMPA recept or antagoni likest ES-481 attenuates tumor growt inh subject affls icted with a glioma thereb y producin oneg or more of the follow ingbenefici aleffects: (a) an alleviation of one or more symptom of the glioma; (b) a dela yor slowing of glioma tumor growt and/oh metastasir s; (c) a stabilized state of a glioma tumo orr malignancy; and/or (d) an increased lifespan as compared to that expected in the absence of treatment. id="p-160" id="p-160" id="p-160" id="p-160" id="p-160" id="p-160"

[0160] Surprisingl they, data further indicated that combining the treatment of ES-481 with temozolomide, an alkylati agentng widely used in the treatment of recurrent malignant gliomas, significant andly drastical enhancedly the tumo growtr atth enuati effectson of ES-481 alone. Indeed, all of the animal streat edwith the combination of ES-481 with temozolomi de 40 survived the entire observati periodon of 40 days and none of these animal shad any measurabl tumoe weir ght.

References: 1 Louis, D.N., etaL (2016). "The 2016 World Health Organizati Classion ficat ofion Tumors of the Central Nervous System a: summary." Acta Neuropathol Published. online 09: May 2016.

DOI 10.1007/500401-016-1545-1. 2 Kerkhof, M. and Vecht C.J. (2013). "Seizure characteristics and prognostic factors of gliomas." Epilepsi a 54(Suppl 9): 12-17. 3 Isselbacher, et al. (1996). Harrison's Principles of Internal Medicine, 13 ed., 1814-1882. 4 The Merck Manual of Diagnosis and Therapy, (2011). 19th Edition, published by Merck Sharp & Dohme Corp., (ISBN 978-0-911910-19-3).

The Encyclopedia of Molecular Cell Biology and Molecular Medicine, Robert S. Porter et al. (eds.) ,published by Blackwel Sciencel Ltd. ,1999-2012 (ISBN 9783527600908). 6 Molecular Biology and Biotechnology: a Comprehensive Desk Reference, (1995). Robert A.

Meyer s(ed.), published by VCH Publishers, Inc. (ISBN 1-56081-569-8). 7 Immunology, (2006). Werner Luttmann, published by Elsevier. 8 Janeway's Immunobiology, (2014). Kenneth Murphy, Allan Mowat, Casey Weave r(eds.) ,Taylor & Francis Limited, (ISBN 0815345305, 9780815345305). 9 Lewin's Genes XI, (2014). published by Jones & Bartl ettPublishers (ISBN-1449659055).

Michael Richar dGree nand Joseph Sambrook, (2012). Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laborator Press,y Cold Spring Harbor, N.Y., USA (ISBN 1936113414). 11 Davis et al., (2012). Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (ISBN 044460149X). 12 Laboratory Methods in Enzymology: DNA, (2013). Jon Lorsch (ed.) Elsevier (ISBN 0124199542). 13 Current Protocols in Molecular Biology (CPMB), (2014). Frederick M. Ausubel (ed.), John Wiley and Sons (ISBN 047150338X, 9780471503385). 14 Current Protocols in Protein Science (CPPS), (2005). John E. Coligan (ed.), John Wiley and Sons, Inc.

Current Protocols in Immunology (CPI) (2003). John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach Warr, en Strobe (ed, s.) John Wiley and Sons, Inc. (ISBN 0471142735, 9780471142737). 16 Gill ,M.B. and Bredt, D.S. (2011). "An Emerging Role for TARPs in Neuropsychiatric Disorders." Neuropsychopharmacology 36(1): 362-363. 17 Michael A. Rogawski. (2011). "Revisiting AMPA Receptors as an Antiepileptic Drug Target." Epilepsy Current 11.2.s 18 Louis, D.N., et al. (2016). "The 2016 World Health Organizati Classion ficat ofion Tumors of the Central Nervous System a: summary." Acta Neuropathol Published. online 09: May 2016.

DOI 10.1007/500401-016-1545-1. 41 19 Levin, A. L., G. E. Shelin, and P. H. Gutin. (1989). "Neoplasms of the central nervous system." In: Devit aS., Hellma n,S., Rosenberg, S. A., editor Cs.ancer: Principles and Practice of Oncology. 3rd Ed., Philadelphia Pa.:, Lippincott; p. 1557-1611.

Kleihues, P., and H. Ohgaki. (1999). "Primary and secondary glioblastomas: from concept to clinical diagnosis." Neuro-Oncol. 1:44-51. 21 Schoenberg, B. S. (1983). "Epidemiology of central nervous system tumor." In: Walker, M D, editor.

Oncology of the Nervous System. Boston: Nijhoff; p. 1-30. 22 van Breemen MS, Wilms EB, Vecht CJ. (2007). "Epilepsy in patients with brain tumours: epidemiology, mechanisms, and management." Lancet Neurol 6:42., 1-430. 23 Berendsen S, Varkil M,a Kroonen J, et al. (2016). "Prognostic relevance of epilepsy at presentation in glioblastoma patients." Neur oOncol., 18:700-706. 24 Wirsching, H.G. and Weller M., (2020). "Does Neuronal Activity Promote Glioma Progression?" Trends Cancer, 6(1 ):1-3.

Venkataraman! V., et al. (2019). "Glutamatergic synaptic input to glioma cells drives brain tumour progression." Natur 573:e 532-538. 26 Venkatesh H.S., et al. (2019). "Electrical and synaptic integration of glioma into neural circuits." Nature 573: 539-545. 27 Chong, D.J. and Lerman, A.M. (2016). "Practice Update: Review of Anticonvulsant Therapy." Curren t Neurology and Neuroscience Repor ts16(4): 39. 28 Izumoto S., et al. (2019). "Response to seizure and tumor-progression by treatment of perampanel in patients with gliomas." Neuro-Oncolo Advancgy es, Abstract ACT-13: ii 14. 29 Mayer J., et al. (2020). "Perampanel attenuates epileptiform phenotype in C6 glioma." Neurosci Lett. :715.

FDA Fycompa Label Tablets and Oral Suspension Labe lupdate Aprild 2016, accessed August 2016. 31 Id. 32Id. 33 Venkatesh, H.S., et al. (2019). "Electrical and synaptic integration of glioma into neural circuits." Nature, 573: 539-545. 34 Venkataraman!, V., et al. (2019). "Glutamatergic synaptic input to glioma cells drives brain tumors." Nature, 573: 532-538.

Venkatesh, H.S., et al. (2015). "Neuronal activity promotes glioma growth through neuroligin-3 secretion." Cell, 161(4): 803-816. 36 Venkatesh, H.S., et al. (2017). "Targeting neuronal activity-regulated neuroligin-3 dependency in high­ grade gliomas." Nature, 549: 533-537. 37 Venkatesh, H.S., et al. (2019). "Electrical and synaptic integration of glioma into neural circuits." Nature, 573: 539-545. 38 https://www.atcc.org/Products/AI I/CR.aspxL-2611 39 https://www.atcc.org/products/all/HTB-14.aspx 40 Allen M, et al. (2016). "Origin of the U87MG glioma cell line: Good news and bad news." Sci. Trans.

Med. 8(354): 1-4. 41 Westermark B, Ponten J. Hugosson R. (1973). "Determinants for the establishment of permanent tissue culture lines from human gliomas." Acta Pathol Microbiol.. Scand. A. 81:791-805. 42 42 U.S. Patent No. 8,765,960. 43 Sangr aM, Thorp N, May P, Pizer B, Mallucci C. (2009). "Management strategies for recurrent ependymoma in the paediatric population." Child sNerv. Syst. 25:1283-1291. id="p-161" id="p-161" id="p-161" id="p-161" id="p-161" id="p-161"

[0161] All patents and othe publir cations; includi ngliterature references, issued patents, publishe patentd applications and, co-pending paten applt ications cited; throughout this applicati onare expressly incorporat hereied nby reference for the purpose of describing and disclosing for, exampl e,the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solel fory thei disclosurer prior to the filing date of the present applicati on.Nothing in thi sregard should be construed as an admission that the inventor ares not entitle tod antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representati on as to the content ofs these document iss based on the informatio availablen to the applicants and does not constitut anye admission as to the correctness of the dates or content ofs thes e documents. id="p-162" id="p-162" id="p-162" id="p-162" id="p-162" id="p-162"

[0162] The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the present aspects and embodiments The. present aspects and embodiment ares not to be limite din scope by examples provide d,since the examples are intended as a single illustra tionof one aspect and other functional equily valen embodimentt s are within the scope of the disclosure. Various modifications in addition to those shown and described herein wil lbecome apparent to those skilled in the art from the foregoing descripti on and fall within the scope of the appended claims. The advantages and objects described herein are not necessaril encomy passe byd each embodiment. Those skilled in the art wil lrecognize, or be able to ascerta usingin no more than routin experimente ati manyon, equivalent to sthe specifi embodimentc describeds herein. Such equivalent ares intende tod be encompassed by the follow ingclaims. 43

Claims (8)

CLAIMS CLAIMED IS:

1. A method for treating a malignant glioma in a subject, the method comprising administering to the subject a pharmaceutical composition comprising an effective amount of a selective TARP y8 dependent AMPA receptor antagonist.

2. The method of claim 1, wherein administration of the compound to the patient results in one or more of: (a) an alleviation of one or more symptom of the glioma; (b) a delay or slowing of glioma tumor growth and/or metastasis; (c) a stabilized state of a glioma tumor or malignancy and (d) an increased lifespan as compared to that expected in the absence of treatment.

3. The method of claim 1 or 2, wherein administration of the malignant glioma is selected from the group consisting of: astrocytoma, glioblastoma, oligodendrocytoma, pilocytic astrocytoma, diffuse intrinsic pontine glioma, ependymoma, oligo-astrocytoma, oligodendrogliocytoma, optic pathway glioma, and hypothalamic glioma.

4. A method for treating a malignant glioma in a subject, the method comprising administering to the patient a pharmaceutical composition comprising an effective amount of Formula I: HO—\ CH3 I or a pharmaceutically acceptable salt thereof.

5. A method for treating a malignant glioma in a subject, the method comprising administering to the patient a pharmaceutical composition comprising an effective amount of Formula II: 44 WO 2021/188431 PCT/US2021/022356 II or a pharmaceutically acceptable salt thereof, wherein X is CH or N; A is ; and R1 is selected from the group consisting of: hydrogen, deuterium, fluoro, methyl, HO-(C1-C4 )-alkyl, optionally substituted with one or two methyl or deuterium groups, HO-(C1-C3)-alkoxy, optionally substituted with one or two methyl or deuterium groups, fluoro-(C1-C3)-alkyl, HO-(C1-C3)-alkoxy-methyl, optionally substituted with one or two methyl groups, cyano-(C1-C3)-alkoxy, HO-(C1-C3)-alkylthio, optionally substituted with one or two methyl groups, HO-(C1-C3)-alkyl-NH-, HO-(C1 -C3)-alkyl-N(CH3)-, methylsulfinyl, acyl, aminocarbonyl, 45 WO 2021/188431 PCT/US2021/022356 methylcarbonylmethoxymethyl, aminomethylcarbonyloxyethoxy, triazolylmethyl, 1 -methyl-imidizol-2-ylthio, 5-hydroxymethyl-tetrahydrofuran-2-yl, 3-hyd roxy-3-methylazetid in-1 -yl, 3-methoxy-azetidin-1 -yl 3-methoxy-3-methylazetidin-1 -yl 4-hydroxypiperidin-1-yl, 4-hydroxy-4-methyl-piperidin-1-yl, 4-hydroxy-4-vinyl-piperidin-1-yl, 4-hydroxymethyl-piperidin-1-yl, 4-(2-hydroxyethyl)-piperindin-1-yl, morpholin-4-yl, 2-hydroxymethyl-morpholin-4-yl, morpholin-4-yl-ethoxy, and tetrahydropyran-4-yl, provided that when A is then R1 is not unsubstituted HO-(C1-C3)-alkoxy, deuterium substituted HO-(C1- C3)-alkoxy, or HO-(C1-C3)-alkythio.

6. The method of any one of claims 4-5, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers, diluents, or excipients.

7. The method of any one of claims 1-6, wherein the administration of the pharmaceutical composition is done concomitantly or sequentially with one or more of a surgical resection, chemotherapy, radiotherapy, antiangiogenic therapy, immune therapy, gamma knife radiosurgery, and symptomatic management with corticosteroids. 46 WO 2021/188431 PCT/US2021/022356

8. The method of any one of claims 1-6, wherein the administration of the pharmaceutical composition is done concomitantly or sequentially with the administration of temozolomide. 47