Patent Application
20240066038
Wnt proteins are secreted glycoproteins which bind to cell surface receptors to activate β-catenin-dependent (canonical) and -independent (non-canonical) signaling pathways. A hallmark of the activation of Wnt/β-catenin signaling is the stabilization of cytosolic β-catenin, which enters the nucleus and subsequently binds to transcription factors of the T cell factor/lymphoid enhancing factor (TCF/LEF) family to induce the expression of specific target genes. In the absence of Wnt ligands, β-catenin is phosphorylated by a supramolecular complex containing adenomatous polyposis coli (APC), axin, and glycogen synthetase kinase 3β (GSK3β), and phosphorylated β-catenin becomes multi-ubiquitinated (Ub) and is degraded by the 26S proteasome. However, when Wnt proteins interact with both LRP6 and Fzd, signaling from the cell surface proceeds through the proteins dishevelled (Dvl), axin and GSK3β, resulting in the phosphorylation of LRP6 cytoplasmic tail, the inhibition of GSK3β and the stabilization of cytosolic β-catenin stabilization [1, 2].
In the brain, Wnt/β-catenin signaling is not only crucial for neuronal survival and neurogenesis, but it plays important roles in regulating synaptic plasticity, neuroinflammation, angiogenesis and blood-brain barrier integrity and function [3-5]. While the Wnt/β-catenin pathway is tightly regulated in the adult brain to maintain neurovascular integrity and CNS functions, it is frequently dysregulated in CNS injuries [3-5]. Therefore, restoring Wnt/βcatenin signaling represents a unique opportunity for the rational design of treatment for CNS injuries.
Provided herein are compounds that can modulate Wnt, and can be used to treat central nervous system injuries. In particular, compounds for use in the disclosed methods are as disclosed in Tables A or B, or have a structure of Formula (I) or (I′):
wherein ring B-R2 is
L1 is NH—CO—C0-3alkylene or CO—NH—C0-4alkylene; ring A is a 4-12-membered monocyclic, bicyclic, bridged, or spiro heterocycle comprising a nitrogen ring atom; each R1 is independently H, C1-6alkyl, halo, C1-6haloalkyl, C1-3alkylene-O—C0-3alkylene-C3-C8carbocycle, C0-3alkylene-3-8-membered heterocycle, C0-3alkylene-5-7-membered heteroaryl, or C0-3alkylene-C6-10aryl; R2 is H, F, OH, OMe, or NH2; each X is independently NH2, NMe2, F, or CF3, m is 1 or 2, and n is 1, 2, or 3, with the proviso that when ring A comprises piperidinyl, at least one R1 is other than H.
In some cases, the CNS injury is a stroke, or more specifically, an ischemic stroke or a hemorrhagic stroke. In some cases, the CNS injury is a traumatic brain injury. In some cases, the CNS injury is a spinal cord injury. In some cases, the CNS injury is a cerebral small vessel disease.
Provided herein are methods of treating a central nervous system (CNS) injury by administering to a subject suffering therefrom a Wnt modulator as disclosed herein. In brain, Wnt/β-catenin signaling is not only crucial for neuronal survival and neurogenesis, but it plays important roles in regulating synaptic plasticity, neuroinflammation, angiogenesis and blood-brain barrier integrity and function [3-5]. While the Wnt/β-catenin pathway is tightly regulated in the adult brain to maintain neurovascular integrity and CNS functions, it is frequently dysregulated in CNS injuries [3-5]. Therefore, restoring Wnt/βcatenin signaling represents a unique opportunity for the rational design of treatment for CNS injuries.
Stroke: Stroke is the No. 5 cause of death and a leading cause of long-term disability in adults in the United States. Early management of the poststroke complications in acute stroke patients is critical for preventing further brain damage or promoting repair. Due to the time-limited treatment window, thrombolytic therapy with tissue plasminogen activator (tPA), the only FDA approved pharmacological intervention, is limited to 3%-5% of acute stroke patients. Accumulating evidence suggests that substantial functional improvement after stroke can be achieved with subacutely pharmacological therapies [6].
Ischemic stroke, which makes up about 85% of stroke, occurs when the cerebral blood flow (CBF) is abruptly blocked by an embolus or thrombus. Mounting evidence indicates that activation of Wnt/β-catenin signaling may prevent, ameliorate, or even reverse the negative effects of ischemic brain injury [4, 5, 7]. Indeed, up-regulation of Dkk1, an antagonist of the Wnt/β-catenin pathway by binding to the Wnt co-receptor LRP6, and downregulation of Wnt/β-catenin signaling were detected within the ischemic region as early as 3 h after middle artery occlusion (MCAo) [8]. Moreover, Dkk1 level is significantly increased in peripheral blood of patients with acute ischemic stroke or with stable cerebrovascular disease [9, 10]. Importantly, intranasal Wnt3A displayed significantly neuroprotective and regenerative effects after focal ischemic stroke in mice and rats [11, 12]. Interestingly, most of the beneficial effects of Wnt3a were abolished after intranasal administration of Wnt antagonist Dkk1 or small molecule Wnt inhibitors XAV-939 [11]. Together, these studies indicate that the Wnt/β-catenin pathway is an attractive therapeutic target to promote neurovascular repair following ischemic stroke.
Hemorrhagic stroke, which makes up about 15% of stroke cases, is a devastating pathological condition as it is likely to cause fatality or severe disability in survivors. Studies have demonstrated that Wnt/β-catenin signaling plays a critical role in disease pathogenesis of hemorrhagic stroke [4, 7]. Particularly, the Wnt/β-catenin signaling pathway is greatly inhibited as early as 6 h after onset of hemorrhagic stroke [13, 14], and remains downregulated up to 2 weeks after onset [4]. Importantly, exogenous intranasal delivery of Wnt3A into the rat brain with subarachnoid hemorrhage (SAH) alleviated neuronal apoptosis, improved the neurological scores, brain water content and long-term neurobehavioral functions after SAH, and these beneficial effects was totally abolished after administration of Fzd-1 siRNA [14]. Similarly, exogenous delivery of recombinant human Wnt1 into the ventricle of SAH rats alleviated subarachnoid hemorrhage-induced early brain injury, which was abolished after administration of Wnt1 siRNA or a neutralizing monoclonal antibody anti-
Fzd1 [13]. Together, these studies indicate that the Wnt/β-catenin pathway is an attractive therapeutic target to promote neurovascular repair following hemorrhagic stroke.
Traumatic brain injury: Traumatic brain injury (TBI) is an injury to the brain caused by a physical force, leading to temporary or permanent impairment in cognitive, physical, and psychosocial functions [15]. Numerous studies have demonstrated that Wnt/β-catenin signaling plays an important role in TBI pathobiology and is a novel target for intervention in secondary injury after TBI [4, 7]. It has been reported that Dkk1 level is elevated in the serum of patients with severe TBI, and is closely associated with increasing severity of the trauma and higher risk of short-term mortality [16]. Moreover, Wnt3A given via intranasal administration or intravenous injection improves functional recovery after TBI by modulating autophagic, apoptotic, and regenerative pathways in the mouse brain [17, 18]. Therefore, Wnt/β-catenin signaling plays an important role in TBI pathobiology, and this pathway is a novel target for intervention in secondary injury after TBI [4, 7].
Spinal cord injury: Spinal cord injury (SCI) is a sudden onset disruption to the neuronal tissue within the spinal canal and can be classified into primary and secondary injuries. While the primary injury is an irreversible damage caused by the initial traumatic event, the secondary injury is created by a series of biological and functional changes including spinal cord hemorrhage, edema, ischemia reperfusion, apoptosis, inflammatory reaction, etc [19]. Although Wnts are barely expressed in uninjured spinal cord, exogenous Wnt3a administration is able to promote axon conduction and regeneration, reduce neuronal cell death, and ultimately enhance the recovery of neurological function after SCI [20-22].
Cerebral small vessel disease: Cerebral small vessel disease (CSVD) is refers to different pathological processes that affect the small vessels of the brain, including small arteries, arterioles and capillaries, and can cause stroke and dementia, mood disturbance and gait problems [23, 24]. Blood-brain barrier (BBB) damage is a critical pathological feature of CSVD [24]. Studies in past ten years have established that the Wnt/β-catenin pathway is a key pathway required not only for BBB formation but also for BBB integrity and function [3, 24-26]. Indeed, dysregulation of Wnt/β-catenin may limit remyelination of white matter lesions (WMLs) and render the BBB leaky [24, 27, 28]. Therefore, restoring Wnt/β-catenin signaling brings therapeutic opportunities to repair BBB damages and reduce white matter injuries in CSVD.
As used herein, the terms “treat,” “treating,” “treatment,” and the like refer to eliminating, reducing, or ameliorating a disease or condition, and/or symptoms associated therewith. Although not precluded, treating a disease or condition does not require that the disease, condition, or symptoms associated therewith be completely eliminated. As used herein, the terms “treat,” “treating,” “treatment,” and the like may include “prophylactic treatment,” which refers to reducing the probability of redeveloping a disease or condition, or of a recurrence of a previously-controlled disease or condition, in a subject who does not have, but is at risk of or is susceptible to, redeveloping a disease or condition or a recurrence of the disease or condition. The term “treat” and synonyms contemplate administering a therapeutically effective amount of a compound as disclosed herein to an individual in need of such treatment.
The term “treatment” also includes relapse prophylaxis or phase prophylaxis, as well as the treatment of acute or chronic signs, symptoms and/or malfunctions. The treatment can be orientated symptomatically, for example, to suppress symptoms. It can be effected over a short period, be oriented over a medium term, or can be a long-term treatment, for example within the context of a maintenance therapy.
The term “therapeutically effective amount,” as used herein, refers to an amount of a compound sufficient to treat, ameliorate, or prevent the identified disease or condition, or to exhibit a detectable therapeutic, prophylactic, or inhibitory effect. The effect can be detected by, for example, an improvement in clinical condition, reduction in symptoms, or by any of the assays or clinical diagnostic tests described herein or known in the art. The precise effective amount for a subject will depend upon the subjects body weight, size, and health; the nature and extent of the condition; and the therapeutic or combination of therapeutics selected for administration. Therapeutically effective amounts for a given situation can be determined by routine experimentation that is within the skill and judgment of the clinician.
Presented herein are methods of administering a compound as disclosed herein or salt thereof as the neat compound or as a pharmaceutical composition orally, intravenously, or parenterally. In some cases, the compound or salt thereof is administered orally. Administration of a pharmaceutical composition, or neat compound, can be performed during or after the onset of the disease or condition of interest. Typically, the pharmaceutical compositions are sterile, and contain no toxic, carcinogenic, or mutagenic compounds that would cause an adverse reaction when administered.
Provided herein are methods of treating CNS injuries by administering a Wnt modulator to a subject suffering therefrom. The Wnt modulator is a compound as discussed below, or a pharmaceutically acceptable salt thereof. The compounds can have a structure of formula (I), or more specifically (I′), or pharmaceutically acceptable salts thereof:
wherein
As used herein, the term “alkyl” refers to straight chained and branched saturated hydrocarbon groups containing one to thirty carbon atoms, for example, one to twenty carbon atoms, or one to ten carbon atoms. The term Cn means the alkyl group has “n” carbon atoms. For example, C4 alkyl refers to an alkyl group that has 4 carbon atoms. C1-6 alkyl refers to an alkyl group having a number of carbon atoms encompassing the entire range (e.g., 1 to 6 carbon atoms), as well as all subgroups (e.g., 2-6, 1-5, 3-6, 1, 2, 3, 4, 5, and 6 carbon atoms). Nonlimiting examples of alkyl groups include, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl (2-methylpropyl), t-butyl (1,1-dimethylethyl), 3,3-dimethylpentyl, and 2-ethylhexyl. Unless otherwise indicated, an alkyl group can be an unsubstituted alkyl group or a substituted alkyl group. A “haloalkyl” group is an alkyl group having at least one halo substituent. In some cases, the haloalkyl comprises 1, 2, or 3 halo substituents, or can comprise a perhaloalkyl (i.e., all hydrogen atoms of the alkyl group are substituted with a halo). Non-limiting examples of haloalkyl include trifluoromethyl, fluoroethyl, difluoroethyl, and trifluoroethyl.
The term “alkylene” used herein refers to an alkyl group having a substituent. For example, an alkylene group can be —CH2CH2— or —CH2— or —CH2CH(CH3)—. The term Cn means the alkylene group has “n” carbon atoms. For example, C1-4 alkylene refers to an alkylene group having a number of carbon atoms encompassing the entire range, as well as all subgroups, as previously described for “alkyl” groups. A C0alkylene group refers to a direct bond. Unless otherwise indicated, an alkylene group can be an unsubstituted alkylene group or a substituted alkylene group.
As used herein, the term “carbocycle” refers to an aliphatic cyclic hydrocarbon group containing three to eight carbon atoms (e.g., 3, 4, 5, 6, 7, or 8 carbon atoms). The term Cn means the carbocycle group has “n” carbon atoms. For example, C5 carbocycle refers to a carbocycle group that has 5 carbon atoms in the ring. C6-C8 carbocycle refers to carbocycle groups having a number of carbon atoms encompassing the entire range (e.g., 6 to 8 carbon atoms), as well as all subgroups (e.g., 6-7, 7-8, 6, 7, and 8 carbon atoms). Nonlimiting examples of carbocycle groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Unless otherwise indicated, a carbocycle group can be an unsubstituted carbocycle group or a substituted carbocycle group.
As used herein, the term “heterocycle” is defined similarly as carbocycle, except the ring contains one to three heteroatoms independently selected from oxygen, nitrogen, and sulfur. In particular, the term “heterocycle” refers to a ring containing a total of three to twelve atoms (e.g., 3-8, 5-8, 3-6, 4-12, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12), of which 1, 2, or 3 of the ring atoms are heteroatoms independently selected from the group consisting of oxygen, nitrogen, and sulfur, and the remaining atoms in the ring are carbon atoms. The heterocycle can be monocyclic, bicyclic, bridged, or spiro heterocycle. In some cases, the heterocycle can be a 4-12 membered ring and comprises only one ring heteroatom—and in particular embodiments, a sole nitrogen ring heteroatom. Nonlimiting examples of heterocycle groups include piperdine, pyrazolidine, tetrahydrofuran, tetrahydropyran, dihydrofuran, morpholine, and the like. In some cases, the heterocycle can be azetidinyl, piperidinyl, pyrollidinyl, decahydroquinolinyl, octahydroindolizinyl, quinuclidinyl, azaspiro[5.5]undecanyl, azabicyclo[2.1.1]hexanyl, azepanyl, or hexahydropyrrolizinyl.
Carbocycle and heterocycle groups can be saturated or partially unsaturated ring systems optionally substituted with, for example, an R1 group as disclosed herein. Heterocycle groups optionally can be further N-substituted with an R1 group, e.g., alkyl (for example, methyl or ethyl), alkylene-carbocycle, alkylene-aryl, and alkylene-heteroaryl.
As used herein, the term “aryl” refers to a monocyclic or bicyclic aromatic group, having 6 to 10 ring atoms. Unless otherwise indicated, an aryl group can be unsubstituted or substituted with one or more, and in particular one to four groups independently selected from, for example, halo, alkyl, alkenyl, OCF3, NO2, CN, NC, OH, alkoxy, amino, CO2H, CO2alkyl, aryl, and heteroaryl. Aryl groups can be isolated (e.g., phenyl) or fused to another aryl group (e.g., naphthyl), a carbocycle group (e.g. tetraydronaphthyl), a heterocycloalkyl group, and/or a heteroaryl group.
As used herein, the term “heteroaryl” refers to a monocyclic or bicyclic aromatic ring having 5 to 10 total ring atoms, and containing one to four heteroatoms selected from nitrogen, oxygen, and sulfur atom in the aromatic ring. Unless otherwise indicated, a heteroaryl group can be unsubstituted or substituted with one or more, and in particular one to four, substituents selected from, for example, halo, alkyl, alkenyl, OCF3, NO2, CN, NC, OH, alkoxy, amino, CO2H, CO2alkyl, aryl, and heteroaryl. In some cases, the heteroaryl is a 5-7 membered monocyclic ring having 1 to four ring heteroatoms. In some cases, the heteroaryl group is substituted with one or more of alkyl and alkoxy groups. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, pyridyl, pyrrolyl, oxazolyl, triazinyl, triazolyl, isothiazolyl, isoxazolyl, imidazolyl, pyrazinyl, pyrimidinyl, thiazolyl, and thiadiazolyl.
In various cases, m is 1. In some cases, m is 1 and X is NH2; and in some specific cases, X is meta to the amide bond on the phenyl ring, and in other specific cases, X is para to the amide bond on the phenyl ring. In some cases, m is 1 and X is NMe2, and in some specific cases, X is ortho to the amide bond on the phenyl ring. In some cases, m is 1 and X is CF3, and in some specific cases, X is meta to the amide bond on the phenyl ring. In some cases, m is 1, and X is F, and in some specific cases, X is ortho to the amide bond on the phenyl ring. In some cases, m is 2. In various cases, m is 2 and one X is NH2and one X is F, and in some specific cases, each X is ortho to the amide bond on the phenyl ring.
In various cases, ring A of Formula (I) or (I′) can comprise azetidinyl, piperidinyl, pyrollidinyl, decahydroquinolinyl, octahydroindolizinyl, quinuclidinyl, azaspiro[5.5]undecanyl, azabicyclo[2.1.1]hexanyl, azepanyl, or hexahydropyrrolizinyl. In some cases, the substituent on ring A, R1, can be fluoro, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, CH2cyclohexyl, CH2cyclopenyl, methyl, CH2CH2OCH3, isopropyl, difluoroethyl, trifluoroethyl, tetrahydropyranyl, CH2-(methyl-isoxazolyl), methyl-pyrazolyl, CH2CH2phenyl, CH2phenyl, CH2(methoxyphenyl), or phenyl.
In various cases, ring B-R2 is
In various cases, ring B-R2 is
In some cases, R2 is H. In some cases, R2 is F. In some cases, R2 is OH, OMe, or NH2.
In various cases, L1 is NHCO—C0-4aklylene. In various cases, L1 is CONH—C0-4aklylene. In some cases, L1 is CONHCH2, or CONHCH2CH2, or CONHCH2CH(CH3).
In various cases, the compound can have a structure of Formula (IA), (IA′), (IA″), (IA′″), (IB), (IB′), (IC), or (IC′):
In some cases, the compound has a structure of Formula (ID) or (ID′):
wherein when L1 is attached to the ring nitrogen, R1 on the ring nitrogen is null. In various cases, the compound has a structure of Formula (IE) or (IE′):
wherein when L1 is attached to the ring nitrogen, R1 on the ring nitrogen is null.
Specific compounds contemplated for use in the present disclosure include those as provided in Table A, below, or a pharmaceutically acceptable salt thereof.
In some cases, the compound of Table A is 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, or 1057, or a pharmaceutically acceptable salt thereof.
In some cases, a compound for use in disclosed methods is as shown in Table B, or a pharmaceutically acceptable salt thereof.
In some cases, the compound for use in the disclosed methods is one of Compounds 1009, 1011, 1013, 1015, 1020, 1045, 1046, 1050, 1023, 1038, 1039, or 1041, or a pharmaceutically acceptable salt thereof.
In some cases, the compound for use in the disclosed methods is one of Compounds 1054, 1055, 1056, or 1057, or a pharmaceutically acceptable salt thereof.
The salts, e.g., pharmaceutically acceptable salts, of compounds disclosed herein can be prepared by reacting the appropriate base or acid with an appropriate amount of compound.
Acids commonly employed to form pharmaceutically acceptable salts include inorganic acids such as hydrogen bisulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include anions, for example sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, O-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate.
Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Pharmaceutically acceptable salts of compounds may also be prepared with a pharmaceutically acceptable cation. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations. Carbonates or hydrogen carbonates are also possible. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium, or ferric, and the like. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,Ndibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.
The compounds can be synthesized as disclosed in, e.g., International Application No. PCT/US2020/41186.
Dosages of the compounds disclosed herein can be administered as a dose measured in mg/kg. Contemplated mg/kg doses of the disclosed compounds include about 0.001 mg/kg to about 1000 mg/kg. Specific ranges of doses in mg/kg include about 0.1 mg/kg to about 500 mg/kg, about 0.5 mg/kg to about 200 mg/kg, about 1 mg/kg to about 100 mg/kg, about 1 mg/kg to about 50 mg/kg, about 1 mg/kg to about 40 mg/kg, and about 5 mg/kg to about 30 mg/kg.
A compound used in a method described herein can be administered in an amount of about 0.005 to about 750 milligrams per dose, about 0.05 to about 500 milligrams per dose, or about 0.5 to about 250 milligrams per dose. For example, a compound can be administered, per dose, in an amount of about 0.005, 0.05, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or750 milligrams, including all doses between 0.005 and 750 milligrams.
As herein, the compounds described herein may be formulated in pharmaceutical compositions with a pharmaceutically acceptable excipient, carrier, or diluent. The compound or composition comprising the compound is administered by any route that permits treatment of the disease or condition. One route of administration is oral administration. Additionally, the compound or composition comprising the compound may be delivered to a patient using any standard route of administration, including parenterally, such as intravenously, intraperitoneally, intrapulmonary, subcutaneously or intramuscularly, intrathecally, topically, transdermally, rectally, orally, nasally or by inhalation. Slow release formulations may also be prepared from the agents described herein in order to achieve a controlled release of the active agent in contact with the body fluids in the gastro intestinal tract, and to provide a substantial constant and effective level of the active agent in the blood plasma. The crystal form may be embedded for this purpose in a polymer matrix of a biological degradable polymer, a water-soluble polymer or a mixture of both, and optionally suitable surfactants. Embedding can mean in this context the incorporation of micro-particles in a matrix of polymers. Controlled release formulations are also obtained through encapsulation of dispersed micro-particles or emulsified micro-droplets via known dispersion or emulsion coating technologies.
Administration may take the form of single dose administration, or a compound as disclosed herein can be administered over a period of time, either in divided doses or in a continuous-release formulation or administration method (e.g., a pump). However the compounds disclosed herein are administered to the subject, the amounts of compound administered and the route of administration chosen should be selected to permit efficacious treatment of the disease or condition.
In an embodiment, the pharmaceutical compositions are formulated with one or more pharmaceutically acceptable excipient, such as carriers, solvents, stabilizers, adjuvants, diluents, etc., depending upon the particular mode of administration and dosage form. The pharmaceutical compositions should generally be formulated to achieve a physiologically compatible pH, and may range from a pH of about 3 to a pH of about 11, preferably about pH 3 to about pH 7, depending on the formulation and route of administration. In alternative embodiments, the pH is adjusted to a range from about pH 5.0 to about pH 8. More particularly, the pharmaceutical compositions may comprise a therapeutically effective amount of at least one compound as described herein, together with one or more pharmaceutically acceptable excipients. Optionally, the pharmaceutical compositions may comprise a combination of the compounds described herein, or may include a second active ingredient useful in the treatment or prevention of a disorder as disclosed herein.
Formulations, e.g., for parenteral or oral administration, are most typically solids, liquid solutions, emulsions or suspensions, while inhalable formulations for pulmonary administration are generally liquids or powders. A pharmaceutical composition can also be formulated as a lyophilized solid that is reconstituted with a physiologically compatible solvent prior to administration. Alternative pharmaceutical compositions may be formulated as syrups, creams, ointments, tablets, and the like.
The term “pharmaceutically acceptable excipient” refers to an excipient for administration of a pharmaceutical agent, such as the compounds described herein. The term refers to any pharmaceutical excipient that may be administered without undue toxicity.
Pharmaceutically acceptable excipients are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there exists a wide variety of suitable formulations of pharmaceutical compositions (see, e.g., Remington Pharmaceutical Sciences).
Suitable excipients may be carrier molecules that include large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactive virus particles. Other exemplary excipients include antioxidants (e.g., ascorbic acid), chelating agents (e.g., EDTA), carbohydrates (e.g., dextrin, hydroxyalkylcellulose, and/or hydroxyalkylmethylcellulose), stearic acid, liquids (e.g., oils, water, saline, glycerol and/or ethanol) wetting or emulsifying agents, pH buffering substances, and the like. Liposomes are also included within the definition of pharmaceutically acceptable excipients.
The pharmaceutical compositions described herein are formulated in any form suitable for an intended method of administration. When intended for oral use for example, tablets, troches, lozenges, aqueous or oil suspensions, non-aqueous solutions, dispersible powders or granules (including micronized particles or nanoparticles), emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents including sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a palatable preparation.
Pharmaceutically acceptable excipients particularly suitable for use in conjunction with tablets include, for example, inert diluents, such as celluloses, calcium or sodium carbonate, lactose, calcium or sodium phosphate; disintegrating agents, such as cross-linked povidone, maize starch, or alginic acid; binding agents, such as povidone, starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc.
Tablets may be uncoated or may be coated by known techniques including microencapsulation to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax may be employed.
Formulations for oral use may be also presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example celluloses, lactose, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with non-aqueous or oil medium, such as glycerin, propylene glycol, polyethylene glycol, peanut oil, liquid paraffin or olive oil.
In another embodiment, pharmaceutical compositions may be formulated as suspensions comprising a compound of the embodiments in admixture with at least one pharmaceutically acceptable excipient suitable for the manufacture of a suspension.
In yet another embodiment, pharmaceutical compositions may be formulated as dispersible powders and granules suitable for preparation of a suspension by the addition of suitable excipients.
Excipients suitable for use in connection with suspensions include suspending agents (e.g., sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia); dispersing or wetting agents (e.g., a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethyleneoxycethanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate)); and thickening agents (e.g., carbomer, beeswax, hard paraffin or cetyl alcohol). The suspensions may also contain one or more preservatives (e.g., acetic acid, methyl or n-propyl p-hydroxy-benzoate); one or more coloring agents; one or more flavoring agents; and one or more sweetening agents such as sucrose or saccharin.
The pharmaceutical compositions may also be in the form of oil-in water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid paraffin, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth; naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids; hexitol anhydrides, such as sorbitan monooleate; and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring or a coloring agent.
Additionally, the pharmaceutical compositions may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous emulsion or oleaginous suspension. This emulsion or suspension may be formulated by a person of ordinary skill in the art using those suitable dispersing or wetting agents and suspending agents, including those mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,2-propane-diol.
The sterile injectable preparation may also be prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer solution, and isotonic sodium chloride solution. In addition, sterile fixed oils may be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids (e.g., oleic acid) may likewise be used in the preparation of injectables.
To obtain a stable water-soluble dose form of a pharmaceutical composition, a pharmaceutically acceptable salt of a compound described herein may be dissolved in an aqueous solution of an organic or inorganic acid, such as 0.3 M solution of succinic acid, or more preferably, citric acid. If a soluble salt form is not available, the compound may be dissolved in a suitable co-solvent or combination of co-solvents. Examples of suitable co-solvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from about 0 to about 60% of the total volume. In one embodiment, the active compound is dissolved in DMSO and diluted with water.
The pharmaceutical composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle, such as water or isotonic saline or dextrose solution. Also contemplated are compounds which have been modified by substitutions or additions of chemical or biochemical moieties which make them more suitable for delivery (e.g., increase solubility, bioactivity, palatability, decrease adverse reactions, etc.), for example by esterification, glycosylation, PEGylation, etc.
In some embodiments, the compounds described herein may be formulated for oral administration in a lipid-based formulation suitable for low solubility compounds. Lipid-based formulations can generally enhance the oral bioavailability of such compounds.
As such, pharmaceutical compositions comprise a therapeutically or prophylactically effective amount of a compound described herein, together with at least one pharmaceutically acceptable excipient selected from the group consisting of medium chain fatty acids and propylene glycol esters thereof (e.g., propylene glycol esters of edible fatty acids, such as caprylic and capric fatty acids) and pharmaceutically acceptable surfactants, such as polyoxyl 40 hydrogenated castor oil.
wherein
wherein when L1 is attached to the ring nitrogen, R1 on the ring nitrogen is null.
wherein when L1 is attached to the ring nitrogen, R1 on the ring nitrogen is null.
or a pharmaceutically acceptable salt thereof.
Wnt/β-catenin signaling plays a critical role in neuronal survival, neurogenesis, synaptogenesis, neuronal plasticity and synaptic plasticity [3-5]. While Wnt/β-catenin signaling is greatly suppressed in central nervous system injuries, restoring Wnt/β-catenin signaling represents a unique opportunity for the rational design of treatment for central nervous system injuries.
The blood-brain barrier (BBB) is a highly selective semipermeable border of endothelial cells that protects CNS neurons from exposure to neurotoxic blood-derived debris, cells and microbial pathogens. Studies in past ten years have established that the Wnt/β-catenin pathway is a key pathway required not only for BBB formation but also for BBB integrity and function [3]. Moreover, BBB breakdown following middle cerebral artery occlusion can be potently rescued in mice in which Wnt/β-catenin is specifically activated in brain endothelial cells [29]. On the other hand, dysregulation of Wnt/β-catenin may limit remyelination of white matter lesions (WMLs) and render the BBB leaky [24, 27, 28]. Compound 1015 is able to activate Wnt/β-catenin signaling in hCMEC/D3 human cerebral microvascular endothelial cells (
The effect of Wnt modulators in neuroprotection and reduction of the infarct in animals is tested using middle cerebral artery occlusion (MCAO) in mice. This model involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the middle cerebral artery territory [30]. Therefore, C57BL6 mice with distal MCAO at 2-3 months of age are given the Wnt modulator daily via intraperitoneal injection (IP) or oral gavage (PO) at 5 mg/kg to 50 mg/kg, 5 injections per week, for 3 weeks. After treatment, the mice are examined by behavioral tests, and then sacrificed for biological analyses on neuronal cell death, neurogenesis, angiogenesis, neuroinflammation, BBB integrity and permeability, and infarct volume.
The effect of Wnt modulators in neuroprotection in animals is tested using subarachnoid hemorrhage (SHA) in rat models. Experimental subarachnoid hemorrhage rat models are induced by injecting autologous blood into the prechiasmatic cistern [31]. Wnt modulator interventions are applied once SAH models are established. Wnt modulators are given daily via intraperitoneal injection (IP) at 5 mg/kg to 50 mg/kg for 2 days. Brain tissue samples are obtained at 48 hours in the different groups, and cortical cell apoptosis, neuroinflammation, brain edema and neurological impairment are examined.
The effect of Wnt modulators in neuroprotection in animals is tested using controlled cortical impact (CCI) of TBI model in mice. The CCI model is one of the most commonly used models of pre-clinical TBI, and the deficits caused by CCI models have been found to mimic neurobehavioral and cognitive deficits, as they are typically seen after human TBI. CCI in mice is induced with a PCI3000 precision cortical impactor (Hatteras Instruments, Cary, NC) and a 2.8-mm diameter impact tip (velocity=3.0 m/sec, depth=0.5 mm, and contact duration=150 msec) [17]. Wnt modulator interventions are applied 2 hours post-TBI, and then are given daily via intraperitoneal injection (IP) or oral gavage (PO) at 5 mg/kg to 50 mg/kg, 5 injections per week, for 4 weeks. After treatment, the mice are examined by behavioral tests on depression, anxiety, impairment of spatial learning and memory, and multiple motor deficits. The mice are then sacrificed for biological analyses on neuronal cell death, neurogenesis and neuroinflammation.
The effect of Wnt modulators in neuroprotection in animals is tested using contusion model of spinal cord injury (SCI) in rats. The contusion model is established with the modified weight-drop method in rats at 11-13 weeks old and weighing between 220 and 250 g as previously described [32, 33]. Wnt modulator interventions are applied immediately after SCI, and then are given daily via intraperitoneal injection (IP) or oral gavage (PO) at 5 mg/kg to 50 mg/kg, 5 injections per week, for 4 weeks. Behavioral assessment is performed prior to SCI at 24 and 72 h after SCI and then weekly for 6 weeks. Specifically, locomotor function is graded using the Basso-Beattie-Bresnahan (BBB) locomotor rating scale [34], and functional recovery of motor locomotion activity is used to evaluate animalsaibility to maintain their body position on an inclined plate [35]. At the end experiment, the rats are then sacrificed for biological analyses on motor neuronal survival, neurogenesis, neuroinflammation, BBB integrity and permeability, angiogenesis, and lesion size after SCI.
This application claims the benefit of priority to U.S. Provisional Patent Application No. 63/134351 filed Jan. 6, 2021, the disclosure of which is incorporated herein by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US22/11456 | 1/6/2022 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63134351 | Jan 2021 | US |
