Berberine compositions with enhanced oral bioavailability and uses thereof

Abstract

Described herein are compositions comprising berberine and a bioavailability enhancer with improved absorption, oral bioavailability and / or efficacy of berberine, and methods of co-administering berberine and a bioavailability enhancer to a subject and uses thereof.

Description

Docket Number: 233013-40106BERBERINE COMPOSITIONS WITH ENHANCED ORAL BIOAVAILABILITY ANDUSES THEREOFCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims benefit under 35 U.S.C. § 119(e) of U.S.Provisional Application having Serial No. 63 / 239,907 filed January 23, 2024, theentire contents of which is / are hereby incorporated by reference herein.TECHNICAL FIELD

[0002] This invention relates generally to compositions comprising berberineand one or more bioavailability enhancers with improved absorption, oralbioavailability and / or efficacy of berberine. Embodiments discussed herein alsogenerally relate to a method of co-administering berberine and one or morebioavailability enhancers to a subject and uses thereof.BACKGROUND

[0003] Berberine, an isoquinoline plant alkaloid that is found in variousmedicinal herbs such as Coptis chinensis Franch, has been used in traditionalChinese medicine with a long history. Berberine possesses diverse pharmacologicaland therapeutic effects on human for a wide range of diseases or conditions such ashypertension, type 2 diabetes, inflammation, cardiac diseases, hyperlipidemia, andgastrointestinal diseases.

[0004] Despite its therapeutic effects and potentials, pharmacological valuesand efficacy of berberine are limited by its poor bioavailability. It has been reportedthat bioavailability of berberine in human is extremely low (0.5%). Low bioavailabilityof berberine is similarly observed in rat; it was reported that about 99.5% ofberberine is eliminated at the gastrointestinal tract, and about 28% of berberine ismetabolized in liver (Drug Metabolism and Disposition, 2010, 38:1779-1784).

[0005] The major factors contributing to the low bioavailability of berberineinclude high intestinal first-pass elimination, interaction with P-glycoprotein mediatedefflux mechanism, rapid metabolism, and high distribution in liver. Berberine and itsmetabolites (mainly thalifendine, berberrubine, demethyleneberberine, andjatrorrhizine, which are active metabolites exhibiting pharmacological effectsDocket Number: 233013-40106comparable to or less than berberine) are primarily distributed in liver andmetabolized extensively in the liver. P-glycoprotein (P-gp) is abundantly expressed inthe intestinal cells; berberine is a substrate of P-glycoprotein and secreted by P-glycoprotein into the intestinal lumen via the P-glycoprotein mediated effluxmechanism. Berberine is also a substrate of Cytochrome P450 (CYP) 2D6 and 3A4which play roles in berberine metabolism in the gastrointestinal tract and / or liver.Further, berberine exhibits a poor absorption because of its low membranepermeability, low solubility in aqueous solution and tendency of self-aggregation inthe gastrointestinal tract. Under physiological conditions, berberine exists mostly inthe ionized form which easily self-aggregates in the acidic environment of thestomach and upper small intestine. It is classified in the biopharmaceuticalclassification system (BSC) as a class III drug which include drugs that arelipophobic and having poor membrane permeability.

[0006] Berberine is usually provided in chloride form (berberine chloride) incommercial products). Below is the chemical structure of

[0007] In standard therapeutic formulations of berberine, includingsupplements, designed for diabetes, high blood pressure and hyperlipidemia,berberine is administered at a dosage of 500 mg, two to three times per day for 2-24months. On the other hand, most of the reported clinical studies showing efficacyused daily dose of 1,500 mg of berberine. While administering berberine in highdoses accounts for the poor bioavailability and improves efficacy, such high dosescan result in undesirable side effects, such as nausea, abdominal pain, bloating,constipation, diarrhea, and / or hypoglycemia. This high-dose approach is also notcost-effective.Docket Number: 233013-40106

[0008] To improve the oral bioavailability of berberine compositions, the presentinvention provides compositions comprising berberine and one or morebioavailability enhancers, as well as methods of co-administering berberine and oneor more bioavailability enhancers. The use of bioavailability enhancers not onlyresults in an improvement in the therapeutic efficacy of berberine by increasing itsoral absorption, but also reduces and eliminates side effects from high doses, aswell as increases cost effectiveness by relying on lower dosage of berberine. Thepresent invention also provides uses of berberine in conjunction with one or morebioavailability enhancers for modulating a disease or a condition in a subject.SUMMARY

[0009] It is an objective of the present invention to provide a compositioncomprising berberine and at least one bioavailability enhancers with improvedabsorption, oral bioavailability and / or efficacy of berberine.

[0010] It is also an objective of the present invention to provide a method ofenhancing absorption, oral bioavailability and / or efficacy of berberine in a subject byco-administering berberine and at least one bioavailability enhancers to the subject.

[0011] It is another objective of the present invention to provide a method ofmodulating a disease or a condition in a subject by co-administering berberine and atleast one bioavailability enhancers to the subject.BRIEF DESCRIPTION OF FIGURES

[0012] FIG. 1 is a table which summarizes the pharmacokinetics in Sprague-Dawley (SD) rats following oral administration of berberine chloride with variousvehicles, according to Example 4.

[0013] FIG. 2 is a table which summarizes the pharmacokinetics in Sprague-Dawley (SD) rats following oral administration of berberine chloride with variousvehicles, according to Example 5.DETAILED DESCRIPTION

[0014] As used herein and in the claims, the terms “comprising” (or anyrelated form such as “comprise” and “comprises”), “including” (or any related formssuch as “include” or “includes”), “containing” (or any related forms such as “contain”Docket Number: 233013-40106or “contains”), means including the following elements but not excluding others. Itshall be understood that for every embodiment in which the term “comprising” (orany related form such as “comprise” and “comprises”), “including” (or any relatedforms such as “include” or “includes”), or “containing” (or any related forms such as“contain” or “contains”) is used, this disclosure / application also includes alternateembodiments where the term “comprising”, “including,” or “containing,” is replacedwith “consisting essentially of” or “consisting of”. These alternate embodiments thatuse “consisting of” or “consisting essentially of” are understood to be narrowerembodiments of the “comprising”, “including,” or “containing,” embodiments.

[0015] For example, alternate embodiments of “a composition comprising A, B,and C” would be “a composition consisting of A, B, and C” and “a compositionconsisting essentially of A, B, and C.” Even if the latter two embodiments are notexplicitly written out, this disclosure / application includes those embodiments.Furthermore, it shall be understood that the scopes of the three embodiments listedabove are different.

[0016] For the sake of clarity, “comprising”, including, and “containing”, andany related forms are open-ended terms which allows for additional elements orfeatures beyond the named essential elements, whereas “consisting of” is a closedend term that is limited to the elements recited in the claim and excludes anyelement, step, or ingredient not specified in the claim.

[0017] “Consisting essentially of” limits the scope of a claim to the specifiedmaterials, components, or steps (“essential elements”) that do not materially affectthe essential characteristic(s) of the claimed invention. In some embodiments, theessential characteristics are the basic and novel characteristic(s) of the claimedinvention. For example, in some embodiments, the essential elements of acomposition of the disclosure can be “Xmg to Ymg” of compound A. Even if thecomposition includes additional excipients, as long as the additional excipients donot materially affect the essential characteristics of the compound, e.g., in compoundA’s ability to bind to XX target or to treat YY disease, then such embodiment that“consists essentially of compound A” still includes compositions with theaforementioned additional excipients.

[0018] As used herein, the singular forms “a”, “an” and “the” are intended toinclude the plural forms as well, unless the context clearly indicates otherwise.Docket Number: 233013-40106Where a range is referred in the specification, the range is understood to includeeach discrete point within the range. For example, 1-7 means 1, 2, 3, 4, 5, 6, and 7.

[0019] As used herein, the term "about" is understood as within a range ofnormal tolerance in the art and not more than ±10% of a stated value. By way ofexample only, about 50 means from 45 to 55 including all values in between. Asused herein, the phrase "about" a specific value also includes the specific value, forexample, about 50 includes 50.

[0020] As used herein and in the claims, a "therapeutically effective amount",is an amount that is effective to achieve at least a measurable amount of a desiredeffect. For example, the amount may be effective to elicit an immune response,and / or it may be effective to elicit a protective response, against a pathogen bearingthe polypeptide of interest. In some embodiments, the amount may be effective toelicit an immune response against cancer or tumor.

[0021] As used herein, the term "treat," "treating" or "treatment" refers tomethods of alleviating, abating or ameliorating a disease or condition symptoms,preventing additional symptoms, ameliorating or preventing the underlying metaboliccauses of symptoms, inhibiting the disease or condition, arresting the developmentof the disease or condition, relieving the disease or condition, causing regression ofthe disease or condition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition either prophylactically and / ortherapeutically.

[0022] As used herein and in the claims, the term “prevent”, “preventing”,“preventive”, “preventative” or “prevention” refers to the methods of reducing the riskof the onset, relapse or spread of a disease or disorder or one or more of theirsymptoms.

[0023] As used herein, the term “subject” includes a mammal such as ahuman or appropriate non-human mammal, such as primate, mouse, rat, dog, cat,cow, horse, goat, camel, sheep or a pig. And the term “subject in need thereof”,which is interchangeable with the term “subject, refer to a subject having a diseaseor condition, having an increased risk of developing the disease or condition, havingan increased risk of developing such disease or condition relative to the populationat large, or having a refractory or resistant disease disclosed herein, or otherwise beDocket Number: 233013-40106regarded as affected by or manifesting (or likely to be affected by or manifesting)such disease or condition.Bioavailability Enhancer

[0024] In one aspect of this invention, there is provided at least onebioavailability enhancer that is capable of enhancing oral bioavailability and / orefficacy of berberine in a subject by causing directly or indirectly one or more of thefollowings in the subject:a) enhancing the solubility or dissolution rate of berberine;b) enhancing the absorption of berberine;c) reducing the clearance of berberine from gastrointestinal tract or blood;d) reducing the rate or extent of metabolism of berberine in liver;e) reducing the rate or extent of distribution of berberine or its metabolites tonon-target tissues; andf) otherwise enhancing the availability of berberine to the target site of action.

[0025] In one embodiment, the bioavailability enhancer is capable of causingdirectly or indirectly one or more of the following:a) inhibiting or reducing P-glycoprotein mediated efflux transport of berberine;b) inhibiting or reducing the rate or extent of metabolism of berberine or itsmetabolites in liver;c) inhibiting or reducing the rate or extent of distribution of berberine or itsmetabolites to non-target tissues; andd) inhibiting or reducing cytochrome P450 mediated metabolism of berberine orits metabolites in gastrointestinal tract and / or liver.

[0026] In one embodiment, the bioavailability enhancer is a P-glycoproteininhibitor that inhibits or otherwise blocks the function of P-glycoprotein. For example,the P-glycoprotein inhibitor may (i) block the berberine binding site of P-glycoprotein,(ii) interfere with ATP hydrolysis, or (iii) alter integrity of cell membrane lipids aroundthe P-glycoprotein, thereby preventing berberine from being transported out of thecells. Examples of P-glycoprotein inhibitors include, but are not limited to,encequidar (or known as, HM30181), phloretin, biochanin A, baicalein, naringenin,apigenin, ketoconazole, and KX2-4926. Ketoconazole is reported to be a dualinhibitor of P-glycoprotein and CYP3A4. KX2-4926 belongs to a family of compoundsDocket Number: 233013-40106of acetamido-phenylbenzamide derivative which possess P-glycoprotein modulationactivity and / or modulation activity against drug metabolizing enzyme cytochromeP450 (e.g., CYP3A4 and / or CYP3A5 isoforms) as described in WO2022076662A1.Examples of P-glycoprotein inhibitors further include, but are not limited to, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), ketoconazole, alkyl (C12−16)dimethylbenzylammonium chloride (Hyamine), cetyltrimethylammonium bromide(CTAB), magnesium stearate, poly-(ethylene glycol) hexadecyl ether (Brij 58),poly(ethylene oxide)20 sorbitan monolaurate (Tween 20), poly(ethylene oxide)20sorbitanmonooleate (Tween 80), poly(ethyleneoxide)35 modified castor oil(Cremophor EL), polyethylene glycol15-hydroxystearate (Solutol HS 15), sodium 1,4-bis (2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate (AOT), sodiumcaprylate (NaCap),sodium deoxycholate (NaDC), sucrose palmitate (Sisterna 16), sucrose stearate(Sisterna 11), hydrolyzed gelatin (HG), hydroxypropyl cellulose (HPC),hydroxypropylmethyl cellulose (HPMC), poly(ethylene oxide)101-block-poly(propyleneoxide)56-block-poly(ethylene oxide)101 (Pluronic F127), poly(ethylene oxide)80-block-poly(propylene oxide)27-block-poly(ethylene oxide)80 (Pluronic F68), poly(vinylalcohol) (80% hydrolyzed PVA), poly(vinyl alcohol)-graf t-poly(ethylene glycol)copolymer (Kollicoat), poly-(vinylpyrrolidone) (PVP K30), poly-(vinylpolypyrrolidone)(PVPP), poly(ethylene glycol) (PEG), and sodium carboxymethylcellulose (NaCMC).

[0027] In one embodiment, the bioavailability enhancer is a cytochrome P450inhibitor that inhibits or otherwise blocks the function of any subtype of cytochromeP450 enzymes. Examples of cytochrome P450 inhibitors include, but are not limitedto, ketoconazole, KX2-4926 and Sodium Lauryl Sulfate (SLS), Tween 80, Tween 20,amiodarone, amitriptyline, amlodipine, apigenin, aprepitant, armodafinil, artemisinin,aspirin, barbiturates (e.g., amobarbital, pentobarbital, phenobarbital, secobarbital),bergamottin (grapefruit), beta-naphthoflavone, bosentan, buprenorphine, bupropion,cafestol, caffeine, cannabis / cannabidiol, carbamazepine, carvedilol, chloramphenicol,chlorphenamine, chlorpromazine, cimetidine, cinacalcet, ciprofloxacin, citalopram,clemastine, clomethiazole, clomipramine, clopidogrel, clofibrate, clotrimazole,cobicistat, codeine, crisaborole, delavirdine, desipramine, dexamethasone,diphenhydramine, diltiazem, dithiocarbamate, domperidone, donepezil, doxepin,doxorubicin, duloxetine, efavirenz, echinacea, enzalutamide, entacapone,escitalopram, erythromycin, ethanolic compounds (e.g., alcohol), esomeprazole,Docket Number: 233013-40106felbamate, fenofibrate, flavonoids (e.g., amentoflavone (Ginkgo biloba, St. John'sWort), grapefruit), fluvoxamine, fluconazole, fluoxetine, fluvastatin, gabapentin,gemfibrozil, gestodene, glutethimide, halofantrine, haloperidol, hyperforin (St. John'sWort), imatinib, indinavir, indomethacin, insulin, interferon, isoniazid, itraconazole,JWH-018, kava, ketoconazole, lansoprazole, letermovir, letrozole, levomepromazine,lovastatin, memantine, metoclopramide, methadone, methimazole, methoxsalen,metyrapone, mibefradil, midodrine, mifepristone, milk thistle, miconazole,moclobemide, modafinil, montelukast, methylcholanthrene, methylphenidate,metronidazole, naringenin (grapefruit), nefazodone, nelfinavir, niacin (nicotinic acid)and niacinamide (nicotinamide), nicotine, nilutamide, norethisterone, norfloxacin,omeprazole, orphenadrine, pantoprazole, paroxetine, perampanel, perphenazine,phenobarbital, phenylbutazone, phenytoin, pilocarpine, piperine, prednisone,pregabalin, probenecid, promethazine, proton pump inhibitors (e.g., esomeprazole,lansoprazole, omeprazole, pantoprazole, rabeprazole), quinidine, quercetin,ranitidine, ribociclib, rifampicin, rifapentine, risperidone, ritonavir, rucaparib,saquinavir, secobarbital, selegiline, seproxetine ((s)-norfluoxetine / fluoxetinemetabolite), sertraline, simeprevir, sodium valproate (valproic acid), St. John's Wort,sulconazole, sulfamethoxazole, sulfaphenazole, telithromycin, teniposide, terbinafine,teriflunomide, thiotepa, thiazolidinediones, thioridazine, ticlopidine, tioconazole,tobacco, topiramate, tramadol, tranylcypromine, trimethoprim,trimethoprim / sulfamethoxazole, turmeric, valerian, verapamil, voriconazole, Withaniasomnifera (ashwagandha), zafirlukast, and zuclopenthixol.

[0028] In one embodiment, the bioavailability enhancer is a solubilizing agent.Examples of solubilizing agents, but are not limited to, cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin, (2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD), andsulfobutylether-β-cyclodextrin (SBE-β-CD)), water-soluble organic solvents (e.g.polyethylene glycol 300, polyethylene glycol 400, ethanol, propylene glycol, glycerin,N-methyl-2-pyrrolidone, dimethylacetamide, and dimethylsulfoxide), non-ionicsurfactants (e.g., Cremophor EL, Cremophor RH 40, Cremophor RH 60, d-α-tocopherol polyethylene glycol 1000 succinate, polysorbate 20, polysorbate 80,Solutol HS 15, sorbitan monooleate, poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol, Gellucire 44 / 14, Softigen 767, and mono- and di-fatty acid estersof PEG 300, 400, or 1750), water-insoluble lipids (e.g.., castor oil, corn oil,Docket Number: 233013-40106cottonseed oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybeanoil, hydrogenated vegetable oils, hydrogenated soybean oil, and medium-chaintriglycerides of coconut oil and palm seed oil), organic liquids / semi-solids (e.g.,beeswax, d-α-tocopherol, oleic acid, medium-chain mono- and diglycerides), andphospholipids (e.g., hydrogenated soy phosphatidylcholine,distearoylphosphatidylglycerol, l-α-dimyristoylphosphatidylcholine, l-α-dimyristoylphosphatidylglycerol).

[0029] As mentioned above, berberine exists mostly in the ionized form whicheasily self-aggregates in the acidic environment of the stomach and upper smallintestine. One of the ways that a bioavailability enhancer, as provided by thisdisclosure, may enhance the absorption, oral bioavailability and / or efficacy ofberberine is by interfering with the self-aggregation of berberine molecules ormodulating the interactions among berberine molecules. For example, non-ionicsurfactants like Tween 80 and Tween 20 may be used collectively or individually toreduce the aggregation of berberine molecules by direct interaction with hydrophobicpatch regions in the proteins or otherwise.

[0030] In one embodiment, the bioavailability enhancer enhances theabsorption, oral bioavailability and / or efficacy of berberine by inhibiting theelimination of berberine in the gastrointestinal tract. By co-administering berberineand a bioavailability enhancer that inhibits or blocks a P-glycoprotein and / orcytochrome P450, oral bioavailability and hence efficacy of berberine is improved.

[0031] The following experimental results illustrate that oral bioavailability ofberberine is enhanced when berberine is co-administered with a bioavailabilityenhancer, such as P-glycoprotein inhibitors, cytochrome P450 inhibitors, and / orsolubilizing agents. In various examples disclosed herein, the degree of absorptionand oral bioavailability of berberine in a subject is assessed by measuring the AUC(area under the curve) of berberine and its metabolites in the liver is measured. TheAUC of berberine and its metabolites in the liver serves as an important indicator ofthe extent to which the drug escapes intestinal metabolism and reaches the liverintact.Example 1.Docket Number: 233013-40106

[0032] Five compounds which exert P-gp inhibitory effects were screened fortheir ability to enhance oral bioavailability of berberine using Caco-2 cell permeabilityassays. In a Caco-2 cell permeability assay, the transport of a compound ismeasured in the apical to basolateral direction and the basolateral to apical direction.Caco-2 cells (human colorectal adenocarcinoma epithelial cells which express P-gp)mimic the intestinal epithelium and the inhibitory effect of a compound on berberineefflux can be assessed based on the efflux ratio which estimates the magnitude of P-glycoprotein-mediated efflux. Efflux ratio is defined as the ratio of Papp in the B-to-Adirection) / Papp in the A-to-B direction, where B is the basolateral side and A is theapical side. The smaller the efflux ratio, the better the inhibition will be. The resultsin the table below suggest that encequidar was able to attain a low efflux ratio of 1.1,and exhibit much higher potency than the four natural compounds in inhibiting effluxeven at a lower molar ratio (1:0.2) than the latter (1:1).

[0033] Table 1. Efflux Ratios of Berberine in the Presence of P-gp InhibitorsEvaluated using Caco-2 Cell Permeability AssayPap(B-to-A) P (A-tpappo-B) Compound Inhibitor -60- (x 10 Efflux Ratio(x 1 6cm / sec)cm / sec) BerberineNone 1.05 16.9 16.2(10 µM) Encequidar (2 µM) 2.65 3.02 1.1None 0.17 1.92 11.11Phloretin (100 µM) 0.16 1.31 8.21Berberine Biochanin A (100 µM) 0.21 1.32 6.40(100 µM)Baicalein (100 µM) 0.13 0.87 6.48Naringenin (100 µM) 0.20 1.47 7.22Example 2.

[0034] Bioavailability of berberine was examined in rat by studying the AUC ofberberine in liver after berberine (BBR) (200 mg / kg) was administered alone or inconjunction with a P-gp inhibitory agent. The results in below table demonstratedthat encequidar significantly increased the bioavailability of berberine but the otherfour P-gp inhibitors, which are natural P-gp inhibitors, did not. Specifically, co-Docket Number: 233013-40106administration of encequidar (10 mg / kg) with berberine resulted in a 4.2-fold increasein the liver AUC of berberine compared to the vehicle group.

[0035] Table 2. Effects of P-gp Inhibitors on the Liver AUC of Berberine inRats BBR formulation with berberineAUC0-last (ng.h / g) of berberine in liver(200 mg / kg)Control Vehicle (20% DMSO,15% Tween 80, 5% Tween 20,71236 80% Water)Encequidar (10 mg / kg) 301591Phloretin (50 mg / kg) 58546Biochanin A (50 mg / kg) 50517Baicalein (50 mg / kg) 29317Naringenin (50 mg / kg) 45450Example 3.

[0036] Bioavailability of berberine was examined in rat by studying the AUC ofberberine in liver after berberine (200 mg / kg) was administered alone or inconjunction with a P-gp inhibitory agent. In this example, three P-gp inhibitors wereexamined. The AUC of berberine and its metabolites were computed to evaluatetheir distribution in rat liver after the rats taking berberine (200 mg / kg) alone or inconjunction with the P-gp inhibitors. The results in below tables demonstrated thatliver exposure of berberine and its metabolites was enhanced in the presence ofencequidar and KX2-4926, which is likely and predominantly resulted from theenhanced absorption of berberine facilitated by these P-gp inhibitors.

[0037] Table 3. Effect of P-gp Inhibitors on Liver Exposure and Fold-Improvement of Berberine in RatsP-gp inhibitor and BBR Liver exposureBBR doseFold-improvement Groupdose (PO)(PO) (mg / kg)CmaxAUC0-8hin exposure(mg / kg) (ng / g) (ng / g*h)Docket Number: 233013-40106Control Vehicle(9.2% DMSO +0.8% MSA + 90%[13% Tween 80 +7% Kolliphor® EL12497 9896 1+ 80%(30%Propylene glycol200 +70% Water +0.02% conc.HCl)])2 Encequidar (10) 65052 179561 183 KX2-4926 (10) 39492 97855 104 Apigenin (50) 704 3486 0.35

[0038] Table 4. Effect of P-gp Inhibitors on Liver Exposure of BerberineMetabolites in RatsP-gpBBR Liver exposure, AUC0-8h(ng / g*h)BBR inhibitor dose Groupand dose(PO)Berberrubine Demethyleneberberine Jatrorrhizine Thalifendine(PO) (mg / kg) (mg / kg)Docket Number: 233013-40106Propylene glycol +70% Water+ 0.02%conc.HCl)]) Encequidar 23256 1368 13125 56118(10) KX2-4926 32106 1180 8020 35651(10) Apigenin 4534 195 569 3651(50)Example 4.

[0039] The purpose ofstudy was to determine the pharmacokineticcharacteristics of orally-administered berberine chloride following pre-treatment withvarious vehicles to Sprague-Dawley (SD) rats. A total of 160 SD rats were dividedinto 8 groups randomly (20 male rats per group). Each group were single, oralgavage (PO) administered with berberine chloride at a dose equivalent to 200 mg / kgof free berberine following pre-treatment with various vehicles.

[0040] 4.1 Materials

[0041] 4.1.1 Test ArticlesTable 5. Test Articles Used in Example 4Encequidar methanesulfonateTest Article: Berberine chloridemonohydrate Batch / Lot No: B85535 HM-1-C6-2101004

[0042] 4.1.2 VehicleTable 6. Vehicles Used in Example 45.0% (v / v) DMSO (Dimethyl sulfoxide), 28.5% (v / v) propyleneVehicle A: glycol, 66.5% (v / v) DI (deionized) water, 0.02% (v / v) conc. HCl(Hydrochloric acid)Vehicle B: 40.0% (w / v) (2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD) in DIDocket Number: 233013-40106water Vehicle C: 16.0% (w / v) DMSO, 20.0% (w / v) Tween 20, in DI water16.0% (w / v) DMSO, 5.0% (w / v) Tween 20, 15.0% (w / v) TweenVehicle D:80, in DI water16.0% (w / v) DMSO, 15.0% (w / v) Tween 20, 5.0% (w / v) TweenVehicle E:80, in DI water16.0% (w / v) DMSO, 20.0% (w / v) tocopheryl polyethyleneVehicle F:glycol succinate (TPGS), in DI water16.0% (w / v) DMSO, 20.0% (w / v) Tween 20, 1.5% (w / v)Vehicle G:Sodium lauryl sulfate (SLS), in DI waterName Supplier LotDMSO SIGMA SHBR47041, 2-Propanediol (PG) SIGMA STBK9242Deionized distilled Water Wahaha 2110ZJHydrochloric Acid GENERAL P1246465Tween 20 SIGMA-ALDRICH SLCM1935Tween 80 SIGMA WXBD4333VVE-TPGS SIGMA BCCL3146Sodium Dodecyl Sulfate Adamas P2776493HP-β-CD SEEBIO OX2102A

[0043] 4.1.3 Instruments and ReagentsTable 7. Instruments and Reagents Used in the Example 4Catalogue Name VendorNo. SCIEX, Triple-quadrupole MS systemNALiquid chromatography (LC) systemequipped with a binary pump (LC-Shimadzu 30AD), an autosampler (SIL-20ACMP),Co., NA acolumn oven (CTO-20A), a systemJapan controller (CBM-20A) and a degasser(DGU-20A).Docket Number: 233013-40106CatalogueName VendorNo.Milli-Q IQ 7010 Water PurificationMillipore NASystemTissueLyser II Qiagen NAMini-Beadbeater-16 Homogenizer Biospec NAThermoX4R Pro CentrifugeNA ScientificVortex IKA NASemi-automatic sealing Allsheng NACELL CULTURE MICROPLATE, 96Greiner 651180WELL, PS,V-BOTTOMbio-oneCoating Analytical 96B97Pipette (0.5-10 μL) Eppendorf Q60170KPipette (10-100 μL) Eppendorf M10834JPipette (20-200 μL) Eppendorf N17612HPipette (100-1000 μL) Eppendorf M36031IAPRICOT Foil-Seal 4ti-0536 DESIGNS2.0 mL tubes AXYGEN MCT200CSigma-Acetonitrile (HPLC grade)34851-4L Aldrich Sigma- 34860-4L-Methanol (HPLC grade)Aldrich R Sigma-Ammonium acetate431311 AldrichDimethyl sulfoxide (DMSO) (HPLCSigma- 34869 grade) Aldrich JingsuThermo shaker (HT-300)NA XinyunlanPBS Buffer (10x) MCE HY-K3006Sigma- β-glucuronidase 9001-45-0 AldrichDocket Number: 233013-40106Catalogue Name VendorNo. General- Formic acid (HPLC grade)G73553A reagent

[0044] 4.1.4 Animal Acquisition and Assignment to StudyTotal 160 male SD rats (6-8 weeks, 240-280 g) were purchased from JH LaboratoryAnimal Co. Ltd. (Qualification No: 20220009027235). The animals were acclimatedfor at least 5 days before being placed on the study. In all groups, the animals werefasted for at least 12 hours prior to drug administration and were fed 4 hours postberberine chloride dosing. The animals had free access to water at all times.

[0045] 4.2 Methods

[0046] 4.2.1 Dosing and Sampling ScheduleTable 8. Dosing and Sampling Schedule in Example 4Dose Dose Dose Dose Phase GroupTreatment VehicleTime Level Conc. Volume No. (hr) (mg / kg) (mg / mL) (mL / kg) 1Vehicle A Vehicle A -0.5 / / 2Deionized Deionized 2-0.25 / / 8PO-1 water water Berberine Deionized 30 200* 20 10chloride water 1Vehicle A Vehicle A -0.5 / / 2Deionized Deionized 2-0.25 / / 8PO-2 water water Berberine 3Vehicle B 0 200* 20 10chloride 1Vehicle A Vehicle A -0.5 / / 22 Vehicle C Vehicle C -0.25 / / 8PO-3 Berberine 3Vehicle B 0 200* 20 10chloride PO-4 1 Vehicle A Vehicle A -0.5 / / 2Docket Number: 233013-40106Dose Dose Dose Dose Phase GroupTreatment VehicleTime Level Conc. Volume No. (hr) (mg / kg) (mg / mL) (mL / kg) 2Vehicle D Vehicle D -0.25 / / 8Berberine 3Vehicle B 0 200* 20 10chloride 1Vehicle A Vehicle A -0.5 / / 22 Vehicle E Vehicle E -0.25 / / 8PO-5 Berberine 3Vehicle B 0 200* 20 10chloride 1Vehicle A Vehicle A -0.5 / / 22 Vehicle F Vehicle F -0.25 / / 8PO-6 Berberine 3Vehicle B 0 200* 20 10chloride 1Vehicle A Vehicle A -0.5 / / 22 Vehicle G Vehicle G -0.25 / / 8PO-7 Berberine 3Vehicle B 0 200* 20 10chloride 1Encequidar Vehicle A -0.5 10 5 22 Vehicle G Vehicle G -0.25 / / 8PO-8 Berberine 3Vehicle B 0 200* 20 10chloride*The dosage of berberine chloride is 200 mg of free berberine per kg of body weight.

[0047] In this study, SD rats were administered three doses at defined timepoints (-0.5 hours, -0.25 hours, and 0 hours) to investigate the effects ofbioavailability enhancers on the absorption of berberine chloride (equivalent to 200mg / kg free berberine). Groups PO-3~8 were pre-treated with Vehicle A (with orwithout Encequidar) at -0.5 hours, followed by a bioavailability enhancer (Vehicles C,D, E, F, or G) at -0.25 hours, and finally received berberine chloride solubilized inVehicle B (HP-β-CD) at 0 hours. Control Group PO-1 received Vehicle A at -0.5hours, deionized water at -0.25 hours, and berberine chloride suspension preparedin deionized water at 0 hours. Control Group PO-2 followed a similar dosing regimenDocket Number: 233013-40106but received berberine chloride solubilized in Vehicle B (HP-β-CD) instead ofdeionized water at 0 hours. Vehicle B (HP-β-CD) was used as a solubilizing agent forberberine chloride.

[0048] Samples were collected at four time points, specifically at 1, 2, 4, and 8hours post-dose. Terminal bleeding was performed to obtain plasma and liversamples.

[0049] 4.2.2 Dose Formulation Preparation

[0050] The preparation methods for the dosing solutions and vehicles used inthis study are detailed below.

[0051] For PO dosing solution of berberine chloride (Group 2~8) with a targetconcentration of 20 mg / mL (free berberine ion), 10.433 g of berberine chloride and184.995 g of HP-β-CD were accurately weighed into a 500-mL glass bottle. To thismixture, 250 mL of deionized water was added, and the solution was heated to 45°Cwhile stirring with a magnetic stir bar for 15 minutes. The solution was then sonicatedfor 5 minutes and stirred further at 45°C for 90 minutes to achieve a clear,homogeneous, and particle-free yellow solution. The volume was adjusted to462.488 mL with deionized water. The solution was kept stirring at 45°C on a hotplate until use, resulting in a final clear yellow solution at 20 mg / mL.

[0052] For PO dosing solution of berberine chloride (Group 1) with the sametarget concentration (20 mg / mL), 1467.62 mg of berberine chloride was weighedinto a glass bottle. To this, 65.059 mL of deionized water was added, followed bystirring for 40 minutes. The solution was then sonicated for 5 minutes to produce ayellow homogeneous suspension at 20 mg / mL.

[0053] The PO dosing solution for encequidar methanesulfonate monohydrate(5 mg / mL encequidar free base) was prepared by dissolving 114.73 mg ofencequidar methanesulfonate monohydrate in 0.978 mL of DMSO in a glass vial.The solution was vortexed for 2 minutes at room temperature, sonicated for 10minutes, and briefly heated in a 50°C water bath to obtain a completely clear andparticle-free yellow solution. Subsequently, 0.820 mL of this DMSO solution wasadded dropwise into a vial containing 15.580 mL of dilution buffer (4.674 mLpropylene glycol, 10.906 mL deionized water, and 3 µL of 0.02% concentrated HCl).The final mixture was vortexed at room temperature to yield a clear, homogeneous,and particle-free yellow solution at 5 mg / mL.Docket Number: 233013-40106

[0054] For Vehicle A, a solution of 7.36 mL DMSO was added to a 200-mLglass bottle containing 140 mL of dilution buffer (42 mL propylene glycol, 98 mLdeionized water, and 28 µL of 0.02% concentrated HCl). The solution was vortexedat room temperature until it became clear, homogeneous, and particle-free.

[0055] For Vehicle C, in a 100-mL glass bottle, 9.60 g of DMSO wascombined with 12.00 g of Tween 20 and stirred for 5 minutes at room temperatureuntil homogeneous. Deionized water was then added slowly to the 60 mL mark whilestirring. The solution was stirred for 30 minutes at room temperature or until itbecame clear, homogeneous, and particle-free.

[0056] For Vehicle D, a solution was prepared by mixing 9.60 g of DMSO,3.00 g of Tween 20, and 9.00 g of Tween 80 in a 100-mL glass bottle. The mixturewas stirred for 5 minutes at room temperature until homogeneous. Deionized waterwas added slowly to the 60 mL mark while stirring, and the solution was stirred for 30minutes until it became clear, homogeneous, and particle-free.

[0057] For Vehicle E, in a 100-mL glass bottle, 9.60 g of DMSO was combinedwith 9.00 g of Tween 20 and 3.00 g of Tween 80, stirred for 5 minutes at roomtemperature until homogeneous, and diluted with deionized water to the 60 mL mark.The solution was stirred for 30 minutes at room temperature until it became clear,homogeneous, and particle-free.

[0058] For Vehicle F, a solution was prepared by melting 15.00 g of TPGS at50°C to obtain a clear, particle-free liquid. Then, 9.00 g of melted TPGS was addedto 9.60 g of DMSO in a 100-mL glass bottle and stirred for 10 minutes at roomtemperature until homogeneous. Deionized water was added slowly to the 60 mLmark while stirring, and the solution was stirred for 1.5 hours at room temperatureuntil it became clear, homogeneous, and particle-free.

[0059] For Vehicle G, in a 200-mL glass bottle, 19.20 g of DMSO was mixedwith 24.00 g of Tween 20 and stirred for 5 minutes at room temperature untilhomogeneous. Then, 1.80 g of SLS was added, and the solution was stirred foranother 5 minutes until clear and particle-free. Deionized water was added slowly tothe 120 mL mark while stirring, and the solution was stirred for 30 minutes at roomtemperature until it became clear, homogeneous, and particle-free.

[0060] 4.2.3 Sample Collection and StorageDocket Number: 233013-40106

[0061] Plasma Collection: The animals were euthanized via CO2 inhalation.Approximately 500 μL of blood was collected by cardiac puncture during terminalbleeding into pre-chilled polypropylene (PP) tubes containing EDTA-K2 as ananticoagulant. The blood samples were kept on ice and centrifuged at approximately4°C and 3000 g for 15 minutes to separate plasma within 15 minutes of collection.

[0062] Liver Collection: Following blood collection, approximately 1.0 g of livertissue was harvested. To ensure consistency, the same part of the liver wascollected from each animal.

[0063] The plasma and liver samples were rapidly frozen on dry ice andstored in a freezer at -70°C ± 10°C until further processing for LC / MS / MS analysis.

[0064] 4.2.4 Sample Preparation and Analysis

[0065] 4.2.4.1 Preparation of Liver Standards and Quality Controls (STD / QC)

[0066] For preparation of liver STD / QC, all procedures are conducted underlight-protected conditions. Working solutions were prepared in a DMSO:10 mM PBS(1:4) mixture. To prepare STDs, 5 µL of the working solution was mixed with 95 µL ofblank liver homogenates (prepared by homogenizing tissue in 5 volumes (v / w) of 10mM PBS). For enzymatic incubation, 100 µL of blank homogenates or STDs wascombined with 5 µL of β-glucuronidase (EC 3.2.2.31, type B-1 from bovine liver, 20units / µL in 10 mM PBS buffer) and incubated at 37°C overnight (16 hours). Proteinprecipitation was performed by adding 600 µL of IS (Glipizide, 100 ng / mL) inacetonitrile. For double blanks, 600 µL of acetonitrile was added instead. Themixture was shaken for 5 minutes using an IKA shaker and centrifuged at 14,800rpm for 10 minutes at 4°C. Subsequently, 30 µL of the supernatant was transferredto a new plate, mixed with 30 µL of ACN / H2O (1:1), and 0.5 µL of the solution wasinjected into the LC-MS / MS system for analysis.

[0067] 4.2.4.2 Preparation of Liver Tissue Samples

[0068] For liver tissue samples, homogenization was performed by diluting thetissue in 5 volumes (v / w) of 10 mM PBS, resulting in a dilution factor of 6. For non-diluted samples, 100 µL of liver homogenate was aliquoted into 2.0 mL tubes andcombined with 5 µL of β-glucuronidase (prepared as described above), followed byincubation at 37°C overnight (16 hours). After incubation, 600 µL of the internalstandard (Glipizide, 100 ng / mL in acetonitrile) was added to the samples. Themixture was shaken for 5 minutes using an IKA shaker and centrifuged at 14,800Docket Number: 233013-40106rpm for 10 minutes at 4°C. Subsequently, 30 µL of the supernatant was transferredto a new plate, mixed with 30 µL of ACN / H2O (1:1), and 0.5 µL of the solution wasinjected into the LC-MS / MS system for analysis.

[0069] For sample dilution, specific dilution factors were applied as needed.For a dilution factor of 10, 3 µL of the sample supernatant was mixed with 27 µL ofblank supernatant. For a dilution factor of 20, 1.5 µL of the sample supernatant wasmixed with 28.5 µL of blank supernatant. For a dilution factor of 50, 2 µL of thesample supernatant was mixed with 98 µL of blank supernatant. The calibrationcurve for Berberine in rat liver homogenate was established over a concentrationrange of 2–1000 ng / mL.

[0070] 4.2.4.3 Pharmacokinetic Parameter Estimation

[0071] The pharmacokinetic (PK) parameters of Berberine in the liver,including t1 / 2, Cmax, Tmax, AUClast, and AUCINF, were estimated based on theconcentration-time data and dose level using the non-compartmental analysis modelin WinNonlin® Professional 8.2.

[0072] 4.3 Results

[0073] 4.3.1. Dosing Solution Analysis

[0074] The dosing formulations of Berberine from upper, middle and bottomlayers were diluted and further analyzed by HPLC-UV. Nominal doses were used forPK data analysis since the measured dosing concentration results within ±15% ofthe nominal concentration for mean bias. The results were shown in Table 9.

[0075] Table 9. Single PO Dosing Formulations Analysis Results by HPLC-UVNominal Actual ConcentrationMean BiasSamples Conc. Conc. (mg / mL) (%) (μg / mL) (μg / mL) PO-1-TOP 100 97.420PO-1-MID 100 95.9 -3.66PO- 1-BOT 100 95.7PO-2~PO-8-TOP 100 11020PO-2~PO-8-MID 100 109 8.75PO-2~PO-8-BOT 100 108Note:Mean Bias (%) = (Actual Conc. - Nominal Conc.) / Nominal Conc.*100.Docket Number: 233013-40106

[0076] 4.3.2. Clinical Observations

[0077] Observations of abnormal clinical symptoms were described in Table10.Table 10. Clinical Observation Post-DosingAnimal GroupClinical observationNo. #11~#20: Loose feces was observed at 4 hourpost dose.1 #1~#20#16~#20: Loose feces was observed at 8 hourpost dose.#21~#30: Loose feces was observed at 4 hourpost dose.2 #21~#40#26~#30: Loose feces was observed at 8 hourpost dose.#56~#60: Loose feces was observed at 8 hour3 #41~#60post dose.#76~#80: Loose feces was observed at 8 hour4 #61~#80post dose.#87 and #98: Loose feces was observed at 2hour post dose.5 #81~#100#96~#100: Loose feces was observed at 8hour post dose.#106~#120: Loose feces was observed at 2hour post dose.#111~#120: Loose feces was observed at 46 #101~#120hour post dose.#116~#120: Loose feces was observed at 8hour post dose.#127 and #133: Loose feces was observed at2 hour post dose.7 #121~#140#131~#140: Loose feces was observed at 4hour post dose.Docket Number: 233013-40106Animal GroupClinical observationNo. #136~#140: Loose feces and piloerectionwere observed at 8 hour post dose.#147, #149, #154 and #158: Loose feces wasobserved at 2 hour post dose.#151~#160: Loose feces was observed at 4hour post dose.#156~#160: Loose feces, piloerection anddischarge at the mouth were observed at 88 #141~#160hour post dose.#156, #157 and #159: Activity mediumdecreased was observed at 8 hour post dose.#160: Activity severe decreased and loss ofrighting reflex was observed at 8 hour postdose.

[0078] 4.3.3. Pharmacokinetic Results

[0079] The mean PK parameters of Berberine in rat liver following POadministration of berberine chloride are summarized in FIG. 1.

[0080] In control group PO-1, berberine chloride was administered usingdeionized water as a vehicle. Due to its low solubility in aqueous solution, berberinechloride formed a suspension when fed to rats. In control group PO-2, berberinechloride was solubilized in HP-β-CD. A comparison of controls PO-1 and PO-2demonstrated that the use of HP-β-CD as a solubilizing agent for berberine chlorideincreased the liver AUC by 7.3-fold.

[0081] When berberine chloride solubilized in HP-β-CD (Vehicle B) was pre-treated with bioavailability enhancers Vehicles C, D, E and F containing P-gpinhibitory excipients Tween 20, Tween 80, and / or TPGS in various ratios, the liverAUC increased substantially by approximately 30- to 37-fold. The combination ofTween 20 with an additional CYP450 inhibitory excipient SLS (Vehicle G) in PO-7further improved the liver AUC by approximately 10% compared to PO-3 (Tween 20only, Vehicle C). Additionally, as observed in PO-8, pre-treatment with encequidarDocket Number: 233013-40106prior to the administration of bioavailability enhancer Vehicle G followed by berberinechloride in HP-β-CD resulted in a notable enhancement in the liver AUC, with a foldincrease as high as 168-fold.

[0082] The effects of bioavailability enhancer excipients Tween 20, Tween 80,TPGS, and / or SLS on liver AUC were also evaluated by comparing groups PO-3 toPO-7 (administration of Vehicles C, D, E, F, and G, respectively) with the controlgroup PO-2 (berberine chloride solubilized in HP-β-CD, Vehicle B). This comparisonisolates the standalone effects of Vehicles C, D, E, F, and G, as opposed to thecomparison of PO-3 to PO-7 with the control group PO-1 (berberine chloride indeionized water), which illustrates the combined effects of the bioavailabilityenhancers (Vehicles C, D, E, F, and G) and HP-β-CD as a solubilizing agent. Thedata show that Vehicles C, D, E, F, and G increased liver AUC by approximately 4-to 5-fold when compared to PO-2, indicating the individual contributions of theseexcipients. Furthermore, the addition of encequidar, a P-gp inhibitor, furtherincreased AUC by more than 4-fold when combined with the effects of theseexcipients (fold-increase of 23.2 compared to PO-2).

[0083] In summary, the inventors of this study have identified agents and theircombinations that can significantly enhance bioavailability. The use of HP-β-CD as asolubilizing agent in the berberine formulation was shown to markedly improveberberine bioavailability, compared to its insoluble form (suspension in deionizedwater) which is commonly used in commercially available berberine oral formulations.The administration of bioavailability-enhancing excipients, such as Tween 20, Tween80, TPGS, and / or SLS, demonstrated a standalone effect in further increasing AUC.Furthermore, the use of encequidar when combined with other enhancers resulted ina substantial amplification of the liver AUC.Example 5.

[0084] Example 5 is a follow-up study to Example 4, designed to investigatethe PK of berberine chloride administered at a lower dose (at a dose equivalent to 16mg / kg of free berberine, as opposed to 200 mg / kg in Example 4) without the use of asolubilizing agent in combination with various vehicles. At lower concentrations,berberine chloride dissolves more readily in aqueous solutions compared to higherDocket Number: 233013-40106concentrations. This improved solubility in water is hypothesized to contribute toenhanced bioavailability upon oral administration.

[0085] A total of 220 SD rats were divided into 11 groups (20 male rats pergroup). Each group were single, PO administered with berberine chloride at 16mg / kg in various vehicles.

[0086] 5.1 Materials

[0087] 5.1.1 Test ArticlesTable 10. Test Articles Used in Example 5Encequidar methanesulfonateTest Article: berberine chloridemonohydrate Batch / Lot No: B85535 HM-1-C6-2101004

[0088] 5.1.2 VehiclesTable 11. Vehicles Used in Example 55.0% Dimethyl sulfoxide (DMSO), 28.5% propylene glycol,Vehicle A: 66.5% DI (deionized) water, 0.02% conc. Hydrochloric acid(HCl) 1mg / mL solution of 0.80% Tween 20, 0.36% Neusilin, in DIVehicle B1:water (BBR:Tw20:Neu=1:8:3.6)1 mg / mL solution of 0.80% Tween 20, in DI waterVehicle B2:(BBR:Tw20:TPGS=1:8) 1mg / mL solution of 0.60% Tween 20, 0.20% TocopherylVehicle C1: Polyethylene Glycol Succinate (TPGS), 0.36% Neusilin, in DIwater (BBR:Tw20:TPGS:Neu=1:6:2:3.6)1 mg / mL solution of 0.60% Tween 20, 0.20% TPGS, in DIVehicle C2:water (BBR:Tw20:TPGS=1:6:2)1 mg / mL solution of 0.20%Tween 20, 0.60% TPGS, 0.36%Vehicle D1:Neusilin, in DI water (BBR:Tw20:TPGS:Neu=1:2:6:3.6)1 mg / mL solution of 0.20% Tween 20, 0.60% TPGS, in DIVehicle D2:water (BBR:Tw20:TPGS=1:2:6)1 mg / mL solution of 0.80% Sodium lauryl sulfate (SLS), in DIVehicle E2:water (BBR:SLS =1:8)Vehicle F2: 1 mg / mL solution of 0.40% SLS, 0.40%Tween 20, in DI waterDocket Number: 233013-40106(BBR:SLS:Tw20=1:4:4) Name Supplier LotDMSO SIGMA SHBR47041, 2-Propanediol (PG) SIGMA STBK9242Direct-Pure UPRephiLe S21RDB0304(Ultra Pure water)Hydrochloric Acid GENERAL P1246465Tween20 SIGMA-ALDRICH SLCM1935Neusilin UFL2(Magnesium Aluminum) Fuji Chemical NASilicate Ethanol Aladdin J2430252VE-TPGS SIGMA BCCL3146SLS Adamas P2776493

[0089] 5.1.3 Animal Acquisition and Assignment to StudyTotal 220 male SD rats (6-8 weeks, 180-210 g) were purchased from JH LaboratoryAnimal Co. Ltd. (Qualification No: SCXK (SH) 2022-0009 20220009032796).Animals were fasted for at least 12 hours prior to drug administration and fed 4 hourspost berberine) dosing. Animals had free access to water at all times.

[0090] 5.2 Methods

[0091] 5.2.1 Dosing and Sampling Schedule

[0092] In this study, SD rats were administered two doses at designated timepoints (-0.5 hours and 0 hours). This example aims to examine the effects ofbioavailability enhancers on the absorption of berberine chloride at low dose (16mg / kg). All test groups (PO-Groups 2A, 2B, 3A, 3B, 4A, 4B, 5B, 6B, 7A, or 7B) werepre-treated with Vehicle A (with or without Encequidar) at -0.5 hours, followed byberberine chloride in a vehicle (Vehicles B1, B2, C1, C2, D1, D2, E2, or F2) at 0hours. In groups PO-Groups 3A and 4A, where berberine chloride was mixed withVehicles C1 and D1, respectively, their respective vehicles contain Neusilin, acommercially available excipient typically served as a carrier for solid dispersions.The control group (PO-Group 1) received Vehicle A at -0.5 hours and berberinechloride in deionized water at 0 hours. Samples were collected at four time points,Docket Number: 233013-40106Dose Dose Dose Dose Phase GroupTreatment VehicleTime Level Conc. Volume No. (hr) (mg / kg) (mg / mL) (mL / kg) 1Vehicle A Vehicle A -0.5 / / 21 Berberine Deionized 20 16* 1 16Chloride water 1Vehicle A Vehicle A -0.5 / / 22A Berberine Vehicle 20 16* 1 16Chloride B1 1Vehicle A Vehicle A -0.5 / / 22B Berberine Vehicle 20 16* 1 16Chloride B2 1Vehicle A Vehicle A -0.5 / / 23A Berberine Vehicle 20 16* 1 16Chloride C1 1Vehicle A Vehicle A -0.5 / / 23B Berberine Vehicle 20 16* 1 16Chloride C2 1Vehicle A Vehicle A -0.5 / / 24A Berberine Vehicle 20 16* 1 16Chloride D1 1Vehicle A Vehicle A -0.5 / / 24B Berberine Vehicle 20 16* 1 16Chloride D2 1Vehicle A Vehicle A -0.5 / / 25B Berberine Vehicle 20 16* 1 16Chloride E2specifically at 1, 2, 4, and 8 hours post-dose. Terminal bleeding was performed toobtain plasma and liver samples. The dosing schedule is summarized in Table 12.Table 12. Dosing Schedule in Example 5.Docket Number: 233013-401061 Vehicle A Vehicle A -0.5 / / 26B Berberine Vehicle 20 16* 1 16Chloride F2 1Encequidar Vehicle A -0.5 10 5 27A Berberine Vehicle 20 16* 1 16Chloride B1 1Encequidar Vehicle A -0.5 10 5 27B Berberine Vehicle 20 16* 1 16Chloride B2*The dosage of berberine chloride is equivalent to 16 mg of free berberine per kg ofbody weight.

[0093] 5.2.2 Dose formulations preparation

[0094] The preparation methods for the dosing solutions and vehicles used inthis study are detailed below.

[0095] For Vehicle B1, a total of 110 mg of berberine chloride and 800 mg ofTween 20 were added to 100 mL of a 95:5 (v / v) ethanol:deionized (DI) water solutionin a glass container. The mixture was stirred at 25°C for at least 15 minutes until aclear, homogenous yellow solution free of particles was obtained. Subsequently, 363mg of Neusilin UFL2 was added to the solution, and stirring was continued for 5minutes to form a homogenous yellow suspension without large clumps of solids.The suspension was transferred to a rotary evaporator, and the solvent wasremoved under the following conditions: a water bath at 45°C, 50 mbar pressure, 50rpm rotation, and a condenser temperature of approximately 10°C. This process wascarried out for 1 hour, and a mass balance of ~100% was confirmed. The resultingyellow powder was scraped and appeared dry and free-flowing. The recoveredpowder was weighed and dissolved in the appropriate amount of DI water to preparea 1 mg / mL berberine solution (equivalent to 1.1 mg / mL berberine chloride or 12.73mg / mL of the powder). The mixture was stirred at 25°C for 30 minutes until ahomogenous yellow suspension free of large clumps of solids was obtained. Thesuspension was centrifuged at 12,000 g for 5 minutes, and the supernatant wascarefully collected without disturbing the pellet. The dosing concentration ofberberine was analyzed using HPLC-UV.Docket Number: 233013-40106

[0096] For Vehicle B2, a total of 110 mg of berberine chloride and 800 mg ofTween 20 were added to DI water in a glass container, and the final volume wasadjusted to 100 mL. The mixture was stirred at 25°C for at least 30 minutes until aclear, homogenous yellow solution with minimal particles was obtained. The dosingconcentration of berberine was analyzed using HPLC-UV.

[0097] For Vehicle C1, TPGS was prewarmed in a 50°C water bath forapproximately 20 minutes until it became a clear, particle-free liquid. A total of 110mg of berberine chloride, 600 mg of Tween 20, and 200 mg of TPGS were added to100 mL of a 95:5 (v / v) ethanol:DI water solution in a glass container. The mixturewas stirred at 25°C for at least 15 minutes until a clear, homogenous yellow solutionfree of particles was obtained. Subsequently, 363 mg of Neusilin UFL2 was added tothe solution, and stirring was continued for 5 minutes to form a homogenous yellowsuspension without large clumps of solids. The entire suspension was transferred toa rotary evaporator, and the solvent was removed under the following conditions: awater bath at 45°C, 50 mbar pressure, 50 rpm rotation, and a condensertemperature of approximately 10°C. The process was carried out for 1 hour, and amass balance of ~100% was confirmed. The resulting yellow powder was scrapedand appeared dry and free-flowing. The recovered powder was weighed anddissolved in the appropriate amount of DI water to prepare a 1 mg / mL berberinesolution (equivalent to 1.1 mg / mL berberine chloride or 12.73 mg / mL of the powder).The mixture was stirred at 25°C for 30 minutes until a homogenous yellowsuspension free of large clumps of solids was obtained. The suspension wascentrifuged at 12,000 g for 5 minutes, and the supernatant was carefully collectedwithout disturbing the pellet. The dosing concentration of berberine was analyzedusing HPLC-UV.

[0098] For Vehicle C2, TPGS was prewarmed in a 50°C water bath forapproximately 20 minutes until it became a clear, particle-free liquid. A total of 110mg of berberine chloride, 600 mg of Tween 20, and 200 mg of TPGS were added toDI water in a glass container, and the final volume was adjusted to 100 mL. Themixture was stirred at 25°C for at least 30 minutes until a clear, homogenous yellowsolution with minimal particles was obtained. The dosing concentration of berberinewas analyzed using HPLC-UV.Docket Number: 233013-40106

[0099] For Vehicle D1, TPGS was prewarmed in a 50°C water bath forapproximately 20 minutes until it became a clear, particle-free liquid. A total of 110mg of berberine chloride, 200 mg of Tween 20, and 600 mg of TPGS were added to100 mL of a 95:5 (v / v) ethanol:DI water solution in a glass container. The mixturewas stirred at 25°C for at least 15 minutes until a clear, homogenous yellow solutionfree of particles was obtained. Subsequently, 363 mg of Neusilin UFL2 was added tothe solution, and stirring was continued for 5 minutes to form a homogenous yellowsuspension without large clumps of solids. The suspension was transferred to arotary evaporator, and the solvent was removed under the following conditions: awater bath at 45°C, 50 mbar pressure, 50 rpm rotation, and a condensertemperature of approximately 10°C. The process was carried out for 1 hour, and amass balance of ~100% was confirmed. The resulting yellow powder was scrapedand appeared dry and free-flowing. The recovered powder was weighed anddissolved in the appropriate amount of DI water to prepare a 1 mg / mL berberinesolution (equivalent to 1.1 mg / mL berberine chloride or 12.73 mg / mL of the powder).The mixture was stirred at 25°C for 30 minutes until a homogenous yellowsuspension free of large clumps of solids was obtained. The suspension wascentrifuged at 12,000 g for 5 minutes, and the supernatant was carefully collectedwithout disturbing the pellet. The dosing concentration of berberine was analyzedusing HPLC-UV.

[0100] For Vehicle D2, TPGS was prewarmed in a 50°C water bath forapproximately 20 minutes until it became a clear, particle-free liquid. A total of 110mg of berberine chloride, 200 mg of Tween 20, and 600 mg of TPGS were added toDI water in a glass container, and the final volume was adjusted to 100 mL. Themixture was stirred at 25°C for at least 30 minutes until a clear, homogenous yellowsolution with minimal particles was obtained. The dosing concentration of berberinewas analyzed using HPLC-UV.

[0101] For Vehicle E2, a total of 110 mg of berberine chloride and 800 mg ofSLS were added to DI water in a glass container, and the final volume was adjustedto 100 mL. The mixture was stirred at 45°C for at least 30 minutes until a yellow,homogenous solution with few particles was obtained. The suspension wascentrifuged at 12,000 g for 5 minutes, and the supernatant was carefully collectedDocket Number: 233013-40106without disturbing the pellet. The dosing concentration of berberine was analyzedusing HPLC-UV.

[0102] Regarding Vehicle F2, a total of 110 mg of berberine chloride, 400 mgof Tween 20, and 400 mg of SLS were added to DI water in a glass container, andthe final volume was adjusted to 100 mL. The mixture was stirred at 25°C for at least30 minutes until a yellow suspension free of large clumps of solids was obtained.The suspension was centrifuged at 12,000 g for 5 minutes, and the supernatant wascarefully collected without disturbing the pellet. The dosing concentration ofberberine was analyzed using HPLC-UV.

[0103] 5.2.3 Sample Collection and Storage

[0104] Plasma Collection: At designated time points, the animal was manuallyrestrained, and approximately 150 µL of blood was collected via the jugular vein forterminal sampling into pre-chilled polypropylene (PP) tubes containing K2-EDTA.Blood samples were immediately placed on wet ice and subsequently centrifuged at3000 g for 15 minutes at 4°C to separate plasma. Plasma isolation was completedwithin 15 minutes of sample collection.

[0105] Liver Collection: The animals were euthanized using CO₂ asphyxiation.The abdominal cavity was opened using scissors to expose the internal organs. Thecarcass was held in an upright position, allowing the organs to shift forward naturally.The connective tissues were carefully severed, and the liver was excised. Theexcised tissue was rinsed with cold saline, blotted dry on filter paper, and placed intoa pre-weighed screw-top tube. The liver was snap-frozen by submersion in dry ice.

[0106] The plasma and liver samples were rapidly frozen on dry ice andstored in a freezer at -70°C until further processing.

[0107] 5.3 Results

[0108] 5.3.1 Pharmacokinetic Results

[0109] The mean PK parameters of Berberine in rat liver following POadministration of Berberine Chloride in Example 5 are summarized in FIG. 2.

[0110] In test groups PO-2A, 2B, 3A, 4A, 4B, 5B, and 6B, berberine chlorideat 16mg / kg was administered in vehicles containing bioavailability enhancers Tween20, TPGS, and / or SLS in differing molar ratios. In those groups where administeredwith low dose berberine with the enhancers, the liver AUC increased 1.72- to 3.29-fold compared to control group PO-1. Comparisons between paired groups (2A vs.Docket Number: 233013-401062B, 3A vs. 3B, and 4A vs. 4B) were conducted to evaluate the impact of Neusilin, acommonly used solid dispersion carrier, on AUC. No significant differences wereobserved, suggesting that the improvement in berberine absorption is primarilyattributable to the presence of bioavailability enhancers. Consistent with the findingsin Example 4, pre-treatment with encequidar (in Vehicle A) drastically improved theAUC, resulting in a 27.2- to 30.6-fold increase (as opposed to pre-treatment withVehicle A alone).

[0111] In summary, the ability of bioavailability-enhancing excipients used atvarious molar ratio in improving berberine absorption is confirmed in Example 5.Without solubilizing agents, berberine chloride dissolves better in aqueous solutionat low concentrations, which aids its oral bioavailability. As expected, the addition ofNeusilin as a solid dispersion carrier did not yield effects on the AUC, indicating thatthe observed improvements were driven by the bioavailability enhancers. Similar toExample 4, encequidar was demonstrated to be a potent modulator for enhancingbioavailability. Formulation

[0112] In various embodiments, there is provided a composition comprisingberberine and at least one bioavailability enhancer, or their respectivepharmaceutically acceptable derivatives selected from the group consisting of salts,esters, enol ethers, enol esters, acetals, ketals, orthoesters, hemiacetals, hemiketals,acids, bases, and hydrates thereof.

[0113] In one embodiment of the present invention, berberine is provided as asalt form including all the pharmaceutically acceptable salt form. In one embodiment,berberine is provided as a salt form selected from the group consisting of berberinehydrochloride, berberine chloride, berberine sulfate, berberine phosphate, berberineacetate, berberine succinate, berberine bromide, berberine iodide, berberine citrate,berberine lactate, berberine malate, berberine glutamate, and berberine tannate.

[0114] In one embodiment of the present invention, berberine is provided as aberberine derivative. In one embodiment berberine is provided as a berberinederivative selected from the group consisting of berberine alkaloid, berberine base,berberine hydrochloride, berberine, berberrubine, coreximine, tetrahydropalmatine,jatrorrhizine, 13-hydroxyberberine chloride, coralyne, coralyne chloride, 7,8-dihydro-Docket Number: 233013-4010613-methylberberine, dihydroberberine, berberine acetone, 13-alkylberberine,palmatine, 13-benzylberberine, tetrahydroberberine, tetrahydroprotoberberine, 8-cyanodihydroberberine, 8-oxyberberine, dimeric protoberberine alkaloids,demethylated protoberberine alkaloids, quaternary protoberberine alkaloids,protoberberine and protoberberine alkaloids.

[0115] In one embodiment of the present invention, berberine is provided at aconcentration between 0.01% and 50% of the total weight of the composition. In oneembodiment, berberine is provided at a concentration of 5%, 10%, 15%, 20%, 25%,30%, 35%, 45% or 50% of the total weight of the composition.

[0116] In one embodiment of the present invention, the at least onebioavailability enhancer may include one, two, three, four, five, six, seven, eight, nine,ten or more bioavailability enhancer(s). In one embodiment of the present invention,the at least one bioavailability enhancer is a P-glycoprotein inhibitor. Examples of P-glycoprotein inhibitor include, but are not limited to, encequidar, phloretin, biochaninA, baicalein, naringenin, apigenin, ketoconazole, and KX2-4926, d-α-tocopherylpolyethylene glycol 1000 succinate (TPGS), ketoconazole, alkyl (C12−16)dimethylbenzylammonium chloride (Hyamine), cetyltrimethylammonium bromide(CTAB), magnesium stearate, poly-(ethylene glycol) hexadecyl ether (Brij 58),poly(ethylene oxide)20 sorbitan monolaurate (Tween 20), poly(ethylene oxide)80sorbitan monooleate (Tween 80), poly(ethyleneoxide)35 modified castor oil(Cremophor EL), polyethylene glycol15-hydroxystearate (Solutol HS 15), sodium 1,4-bis (2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate (AOT), sodium caprylate (NaCap),sodium deoxycholate (NaDC), sucrose palmitate (Sisterna 16), sucrose stearate(Sisterna 11), hydrolyzed gelatin (HG), hydroxypropyl cellulose (HPC),hydroxypropylmethyl cellulose (HPMC), poly(ethylene oxide)101-block-poly(propyleneoxide)56-block-poly(ethylene oxide)101 (Pluronic F127), poly(ethylene oxide)80-block-poly(propylene oxide)27-block-poly(ethylene oxide)80 (Pluronic F68), poly(vinylalcohol) (80% hydrolyzed PVA), poly(vinyl alcohol)-graf t-poly(ethylene glycol)copolymer (Kollicoat), poly-(vinylpyrrolidone) (PVP K30), poly-(vinylpolypyrrolidone)(PVPP), poly(ethylene glycol) (PEG), and sodium carboxymethylcellulose (NaCMC).In one embodiment, the at least one bioavailability enhancer is a cytochrome P450inhibitor. Examples of cytochrome P450 inhibitors include, but are not limited to,ketoconazole, KX2-4926, Sodium Lauryl Sulfate (SLS), Tween 80, Tween 20,Docket Number: 233013-40106amiodarone, amitriptyline, amlodipine, apigenin, aprepitant, armodafinil, artemisinin,aspirin, barbiturates (e.g., amobarbital, pentobarbital, phenobarbital, secobarbital),bergamottin (grapefruit), beta-naphthoflavone, bosentan, buprenorphine, bupropion,cafestol, caffeine, cannabis / cannabidiol, carbamazepine, carvedilol, chloramphenicol,chlorphenamine, chlorpromazine, cimetidine, cinacalcet, ciprofloxacin, citalopram,clemastine, clomethiazole, clomipramine, clopidogrel, clofibrate, clotrimazole,cobicistat, codeine, crisaborole, delavirdine, desipramine, dexamethasone,diphenhydramine, diltiazem, dithiocarbamate, domperidone, donepezil, doxepin,doxorubicin, duloxetine, efavirenz, echinacea, enzalutamide, entacapone,escitalopram, erythromycin, ethanolic compounds (e.g., alcohol), esomeprazole,felbamate, fenofibrate, flavonoids (e.g., amentoflavone (Ginkgo biloba, St. John'sWort), grapefruit), fluvoxamine, fluconazole, fluoxetine, fluvastatin, gabapentin,gemfibrozil, gestodene, glutethimide, halofantrine, haloperidol, hyperforin (St. John'sWort), imatinib, indinavir, indomethacin, insulin, interferon, isoniazid, itraconazole,JWH-018, kava, ketoconazole, lansoprazole, letermovir, letrozole, levomepromazine,lovastatin, memantine, metoclopramide, methadone, methimazole, methoxsalen,metyrapone, mibefradil, midodrine, mifepristone, milk thistle, miconazole,moclobemide, modafinil, montelukast, methylcholanthrene, methylphenidate,metronidazole, naringenin (grapefruit), nefazodone, nelfinavir, niacin (nicotinic acid)and niacinamide (nicotinamide), nicotine, nilutamide, norethisterone, norfloxacin,omeprazole, orphenadrine, pantoprazole, paroxetine, perampanel, perphenazine,phenobarbital, phenylbutazone, phenytoin, pilocarpine, piperine, prednisone,pregabalin, probenecid, promethazine, proton pump inhibitors (e.g., esomeprazole,lansoprazole, omeprazole, pantoprazole, rabeprazole), quinidine, quercetin,ranitidine, ribociclib, rifampicin, rifapentine, risperidone, ritonavir, rucaparib,saquinavir, secobarbital, selegiline, seproxetine ((s)-norfluoxetine / fluoxetinemetabolite), sertraline, simeprevir, sodium valproate (valproic acid), St. John's Eort,sulconazole, sulfamethoxazole, sulfaphenazole, telithromycin, teniposide, terbinafine,teriflunomide, thiotepa, thiazolidinediones, thioridazine, ticlopidine, tioconazole,tobacco, topiramate, tramadol, tranylcypromine, trimethoprim,trimethoprim / sulfamethoxazole, turmeric, valerian, verapamil, voriconazole, Withaniasomnifera (ashwagandha), zafirlukast, and zuclopenthixol.Docket Number: 233013-40106

[0117] In one embodiment of the present invention, berberine and the at leastone bioavailability enhancer are provided at a molar ratio of about 1:100 to about100:1. In some embodiments, berberine and the at least one bioavailability enhancerare provided at a molar ratio of about 1:5, about 1:10, about 1:20, about 1:25, about1:50, about 1:75, about 1:80, about 1:90, about 1:100, about 5:1, about 10:1, about20:1, about 25:1, about 50:1, about 75:1, about 80:1, about 90:1, or about 100:1. Forexample, if the at least one bioavailability enhancer is Tween 20, berberine andTween 20 may be provided in a molar ratio of 1:8. In another example, if the at leastone bioavailability enhancer is SLS, berberine and SLS may be provided in a ratio of1:8. In another example, if the at least one bioavailability enhancer is Tween 20 andTPGS, berberine:Tween 20:TPGS may be provided in a ratio of 1:6:2, or 1:2:6. Inanother example, if the at least one bioavailability enhancer is Tween 20 and SLS,berberine:Tween 20:SLS may be provided in a ratio of 1:4:4.

[0118] In one embodiment of the present invention, the at least onebioavailability enhancer is Tween 20. In another embodiment of the present invention,the at least one bioavailability enhancer is SLS. In another embodiment of thepresent invention, the at least one bioavailability enhancer is TPGS. In yet anotherembodiment of the present invention, the at least one bioavailability enhancer isTween 20 and Tween 80. In some embodiments, Tween 20 and Tween 80 arepresent in a molar ratio of about 1:100 to about 100:1. In further embodiment, Tween20 and Tween 80 are present in a molar ratio of about 1:5, about 1:10, about 1:20,about 1:25, about 1:50, about 1:75, about 1:80, about 1:90, about 1:100, about 5:1,about 10:1, about 20:1, about 25:1, about 50:1, about 75:1, about 80:1, about 90:1,or about 100:1. In a particular embodiment, Tween 20 and Tween 80 are present ina ratio of about 1:3. In another particular embodiment, Tween 20 and Tween 80 arepresent in a ratio of about 3:1. In another embodiment, the at least one bioavailabilityenhancer is Tween 20 and SLS. In further embodiment, Tween 20 and SLS arepresent in a molar ratio of about 1:5, about 1:10, about 1:20, about 1:25, about 1:50,about 1:75, about 1:80, about 1:90, about 1:100, about 5:1, about 10:1, about 20:1,about 25:1, about 50:1, about 75:1, about 80:1, about 90:1, or about 100:1. In anexemplary embodiment, Tween 20 and SLS are present in a molar ratio of about 1:1.In another exemplary embodiment, Tween 20 and SLS are present in a molar ratio ofabout 1:13. In another embodiment, the at least one bioavailability enhancer isDocket Number: 233013-40106Tween 20 and TPGS. In some embodiments, Tween 20 and TPGS are present in amolar ratio of about 1:5, about 1:10, about 1:20, about 1:25, about 1:50, about 1:75,about 1:80, about 1:90, about 1:100, about 5:1, about 10:1, about 20:1, about 25:1,about 50:1, about 75:1, about 80:1, about 90:1, or about 100:1. In a particularembodiment, Tween 20 and TPGS are present in a molar ratio of 1:3. In anotherparticular embodiment, Tween 20 and TPGS are present in a molar ratio of 3:1. Inone embodiment of the present invention, the at least one bioavailability enhancer isencequidar. In some embodiments, encequidar is administered in combination withother bioavailability enhancers either concurrently with berberine or prior to thecombined administration of berberine and one or more bioavailability enhancers.Examples of such bioavailability enhancers to be co-administered with encequidarinclude, but are not limited to, Tween 20, Tween 80, TPGS and SLS.

[0119] In some embodiments of the present invention, berberine and the atleast one bioavailability enhancer may be administered in conjunction with a soliddispersion carrier. Examples of the solid dispersion carrier include, but are notlimited to, mesoporous silicas (e.g., SYLOID® AL-1FP / 63FP silicas, SYLOID® 244FP silica, SYLOID® XDP silica, and PERKASIL®), and Neusilin. Neusilin is asynthetic amorphous form of Magnesium Alumino-metasilicate, commercially usedfor improvement of the quality of solid formulations. For example, if the at least onebioavailability enhancer is Tween 20, berberine:Tween 20:Neusilin may be providedin a ratio of 1:8:3.6. In another example, if the at least one bioavailability enhancer isTween 20 and TPGS, berberine:Tween 20:TPGS:Neusilin can be provided in a ratioof 1:6:2:3.6 or 1:2:6:3.6.

[0120] In certain embodiments of the present invention, the administration ofberberine and the at least one bioavailability enhancer in combination with theadministration of a solubilizing agent. Examples of the solubilizing agent include, butare not limited to, cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin, (2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD), and sulfobutylether-β-cyclodextrin (SBE-β-CD)), water-soluble organic solvents (e.g. polyethylene glycol 300, polyethyleneglycol 400, ethanol, propylene glycol, glycerin, N-methyl-2-pyrrolidone,dimethylacetamide, and dimethylsulfoxide), non-ionic surfactants (e.g., CremophorEL, Cremophor RH 40, Cremophor RH 60, d-α-tocopherol polyethylene glycol 1000succinate, polysorbate 20, polysorbate 80, Solutol HS 15, sorbitan monooleate,Docket Number: 233013-40106poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol, Gellucire 44 / 14,Softigen 767, and mono- and di-fatty acid esters of PEG 300, 400, or 1750), water-insoluble lipids (e.g., castor oil, corn oil, cottonseed oil, olive oil, peanut oil,peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils,hydrogenated soybean oil, and medium-chain triglycerides of coconut oil and palmseed oil), organic liquids / semi-solids (e.g., beeswax, d-α-tocopherol, oleic acid,medium-chain mono- and diglycerides), and phospholipids (e.g., hydrogenated soyphosphatidylcholine, distearoylphosphatidylglycerol, l-α-dimyristoylphosphatidylcholine, l-α-dimyristoylphosphatidylglycerol). For example,HP-β-CD may be administered with berberine, concurrently with or after thetreatment with the at least one bioavailability enhancer in order to minimize potentialinteractions between HP-β-CD and the bioavailability enhancer. In variousembodiments, HP-β-CD is utilized in a ratio to berberine ranging from about 1:100 toabout 20:1. In specific embodiments, HP-β-CD is present in a ratio to berberine ofabout 1:1, about 1:5, about 1:10, about 1:20, about 5:1, about 10:1, or about 20:1. Ina further embodiment, HP-β-CD is present in a ratio to berberine of about 20:1. Thesolubilizing agent may be administered to the subject with berberine concurrently orat a predetermined interval after the treatment with the at least one bioavailabilityenhancer. In various embodiments, the solubilizing agent and berberine areadministered at least about 0 minutes, 5 minutes, 10 minutes, 15 minutes, about 30minutes, about 45 minutes, about 1 hour, about 2 hours, about 3 hours, about 4hours, about 5 hours, about 6 hours, or more after the treatment with the at least onebioavailability enhancer.

[0121] As mentioned above, berberine exists mostly in the ionized form whicheasily self-aggregates in the acidic environment of the stomach and upper smallintestine. One of the ways that a bioavailability enhancer, as provided by thisdisclosure, may enhance the absorption, oral bioavailability and / or efficacy ofberberine is by interfering with the self-aggregation of berberine molecules ormodulating the interactions among berberine molecules. For example, non-ionicsurfactants like Tween 80 and Tween 20 may be used collectively or individually toreduce the aggregation of berberine molecules by direct interaction with hydrophobicpatch regions in the proteins or otherwise. Another way is to employ solid dispersioncarriers, such as mesoporous materials like Neusilin, which adsorb and disperseDocket Number: 233013-40106berberine molecules effectively, preventing their aggregation. The use of solubilizingagents such as cyclodextrins can also reduce aggregation by increasing berberine'ssolubility, thereby facilitating enhanced absorption.

[0122] In one embodiment, there is provided a dietary supplementcomposition comprising berberin e and one or more bioavailability enhancers, wherethe absorption and / or bioavailability berberine is enhanced in the presence of thebioavailability enhancer(s). In one embodiment of the dietary supplementcomposition, the bioavailability enhancer is a natural compound that exerts inhibitoryeffects on P-glycoprotein (including P-glycoprotein inhibitor excipients) orcytochrome P450 or both.

[0123] In one embodiment of the present invention, berberine and one type ofbioavailability enhancer are provided together according to an embodiment of thisinvention. In another embodiment of the present invention, berberine and two ormore types of bioavailability enhancers are provided together according to anembodiment of this invention.

[0124] In one embodiment, berberine and / or the bioavailability enhancer is anaturally-occurring compound or a synthetic compound.

[0125] In one embodiment, berberine and the bioavailability enhancer areformulated as a composition in a dosage form selected from the group consisting ofa solution, suspension, powder, tablet, capsule, microemulsion, nano-sized dosageform, liposome, micelles, and phospholipid complex.

[0126] In one embodiment, berberine and the bioavailability enhancer areprovided in different dosage forms and co-administered to a subject.

[0127] In some embodiments, berberine is administered to a subject at doseranging from about 0.1 mg to about 5000 mg per day. In further embodiments,berberine is administered to a human subject at a dose ranging from about 10 toabout 150 mg per day, in combination with at least one bioavailability enhancer at apredetermined ratio. In further embodiments, berberine is administered to a humansubject at a dose of about 10, 20, 30, 40, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100mg, 110 mg, 120 mg, 130 mg, 140 mg, or 150 mg per day, in combination with atleast one bioavailability enhancer at a predetermined ratio. For instance, in a studyinvolving rats administered a dose of 16 mg / kg of berberine, the correspondinghuman equivalent dose can be estimated to be approximately 137.4 mg, computedDocket Number: 233013-40106based on a body surface area conversion factor of 1 / 7 and an average humanweight of 60 kg. In other embodiments, berberine is administered to a human subjectat a dose ranging from about 1,000 to about 2,000 mg per day, in combination withat least one bioavailability enhancer at a predetermined ratio. In further embodiments,berberine is administered to a human subject at a dose of around 1,000 mg, 1,100mg, 1,200 mg, 1,300 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,700 mg , 1,800 mg ,1,900 mg, or 2,000 mg per day, in combination with at least one bioavailabilityenhancer at a predetermined ratio. For instance, in a study involving ratsadministered a dose of 200 mg / kg of berberine, the corresponding human equivalentdose can be estimated to be approximately 1714.2 mg, computed based on a bodysurface area conversion factor of 1 / 7 and an average human weight of 60 kg. Insome embodiments, when berberine is administered to a human subject at a doseabove a predetermined limit, such as 10, 20, 30, 40, 50 mg, 60 mg, 70 mg, 80 mg,90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, or 150 mg, a solubilizing agentmay be used for solubilizing berberine in aqueous solution. A person having ordinaryskill in the art would appreciate that the dose may vary within this range upon thespecies of the subject (e.g. human vs rat), dosage form employed, sensitivity of thesubject, and the route of administration.

[0128] In one embodiment, berberine and the bioavailability enhancer are co-administered to a subject one, two or three times per day. In one embodiment,berberine and the bioavailability enhancer are co-administered to a subject every twodays, three days, or other number of days.Co-administration of Berberine and Bioavailability Enhancer as a CombinationTherapy

[0129] In one aspect of this invention, there is provided a method ofenhancing the absorption, oral bioavailability and / or efficacy of berberine in a subjectby co-administering berberine and one or more bioavailability enhancers to thesubject, or their respective pharmaceutically acceptable derivatives. In anotheraspect, there is provided a method of enhancing the absorption, oral bioavailabilityand / or efficacy of berberine, by providing a bioavailability enhancer in conjunctionwith berberine, or their respective pharmaceutically acceptable derivatives.Docket Number: 233013-40106

[0130] In one aspect of this invention, there is provided a method of co-administering berberine and one or more bioavailability enhancers to a subject inneed thereof by providing to the subject berberine and the bioavailability enhancer(s),or their respective pharmaceutically acceptable derivatives.

[0131] In one aspect of this invention, there is provided a method of treating,preventing or modulating a disease or a condition of a subject by co-administering atherapeutically effective amount of berberine and one or more bioavailabilityenhancers or their respective pharmaceutically acceptable derivatives to a subject inneed thereof.

[0132] In one embodiment, berberine and the bioavailability enhancer areformulated and co-administered as a single composition. In one embodiment,berberine and the bioavailability enhancer are formulated and co-administered asdifferent compositions.

[0133] In one aspect of this invention, berberine and one or morebioavailability enhancers are co-administered to a subject in need for treating,preventing, alleviating, regulating, delaying the onset of, or otherwise modulating adisease (or conditions thereof).

[0134] In one aspect of this invention, there is provided various uses ofberberine and one or more bioavailability enhancers for the preparation of amedicament for treatment, alleviation, regulation, prevention, delay of onset, ormodulation of a disease or conditions thereof.

[0135] In one embodiment, the oral bioavailability of berberine in a subject isenhanced by co-administering berberine with the bioavailability enhancer(s), therebypermitting a lower dose of berberine to be used as compared to the dose ofberberine when given alone for achieving the same biological responses or effects inrespect of the development, progress or deterioration of such disease or conditions.

[0136] In one embodiment, the disease or conditions to be treated, prevented,alleviated, regulated, delayed the onset of, or otherwise modulated includes withoutlimitation diabetes, dyslipidemia, metabolic disorders, cardiovascular diseases,hyperlipidemia, fatty liver, inflammation, Alzheimer’s disease, lung diseases (such astuberculosis, lung cancer, and chronic obstructive pulmonary disease), andinfectious diseases including bacterial infection such as infection caused byMethicillin-resistant Staphylococcus aureus (MRSA).Docket Number: 233013-40106

[0137] Berberine provided by the present invention may exert its actions in asubject in various manners. In one embodiment, berberine exerts its actions bymodulating a relevant signaling pathway or expression of a relevant gene leading tothe development, progress or deterioration of the disease or condition concerned. Inanother embodiment, berberine exerts its actions by prohibiting aggregation or post-translation modification of a molecular target (e.g. attenuate hyperphosphorylation oftau which plays a role in the development of Alzheimer’s disease). In oneembodiment, berberine exert its actions by inducing cell death of the affected cellssuch as inducing non-small cell lung cancer apoptosis via the activation of theROS / ASK1 / JNK pathway.

[0138] A person having ordinary skill in the art would appreciate that thepresent invention, including the compositions and methods described herein, may beapplied on any diseases, disorders and conditions on which berberine is known tohave effects, no matter such knowledge exist before, at the time of, or after thisdisclosure is made.Molecular Targets

[0139] In one aspect of this invention, there is provided a method ofmodulating an activity or function of a molecular target in a subject by co-administering berberine and a bioavailability enhancer to a subject in need thereof.

[0140] In one embodiment, the molecular target is selected from the groupconsisting of AMPK, CD36, acetyl-CoA carboxylase (ACC), microsomal triglyceridetransfer protein (MTTP), scavenger receptor class B type 1 (SR-BI), low-densitylipoprotein receptor (LDLR), ATP-binding cassette transporter A1 (ABCA1), SIRT-,endothelial nitric oxide synthetase (eNOS), lectin-like low-density lipoproteinreceptor-1 (LOX-1), PI3K / AKT (PKB), mTOR, JAK2, calcium channels, NFkB, targetgene of endoplasmic reticulum stress, and HIF-1α.

[0141] In one embodiment, the molecular target is a gene or protein related tometabolism such as lipid metabolism. In another embodiment, the molecular target isa gene or protein related to diabetic control.

[0142] In one embodiment, the molecular target is a gene or protein related toantimicrobial mechanism.Docket Number: 233013-40106

[0143] The foregoing description of the invention has been presented forpurposes of illustration and description and is not intended to be exhaustive or tolimit the invention to the precise form disclosed, and obviously many modificationsand variations are possible in light of the above teaching.

[0144] While the present disclosure may be embodied in many different forms,the discussion are presented with the understanding that the present disclosure is anexemplification of the principles of one or more inventions and is not intended to limitany one embodiments to the embodiments illustrated.

[0145] Further advantages and modifications of the above describedcompositions and methods may readily occur to those skilled in the art.

[0146] The disclosure, in its broader aspects, is therefore not limited to thespecific details, representative devices, systems and methods, and illustrativeexamples shown and described above. Various modifications and variations may bemade to the above specification without departing from the scope or spirit of thepresent disclosure, and it is intended that the present disclosure covers all suchmodifications and variations provided they come within the scope of the followingclaims and their equivalents.Numbered Embodiments

[0147] 1. A composition for the treatment, prevention and / or modulation ofa disease or a conditions in a subject, comprising a therapeutically effective amountof berberine and at least one bioavailability enhancer, or their respectivepharmaceutically acceptable derivatives selected from the group consisting of salts,esters, enol ethers, enol esters, acetals, ketals, orthoesters, hemiacetals, hemiketals,acids, bases, and hydrates thereof.

[0148] 2. The composition of embodiment 1, wherein the berberine ispresent as a salt selected from the group consisting of berberine hydrochloride,berberine chloride, berberine sulfate, berberine phosphate, berberine acetate,berberine succinate, berberine bromide, berberine iodide, berberine citrate,berberine lactate, berberine malate, berberine glutamate, and berberine tannate.

[0149] 3. The composition of embodiment 1, wherein the berberine ispresent as a derivative selected from the group consisting of berberine alkaloid,berberine base, berberine hydrochloride, berberine, berberrubine, coreximine,Docket Number: 233013-40106tetrahydropalmatine, jatrorrhizine, 13-hydroxyberberine chloride, coralyne, coralynechloride, 7,8-dihydro-13-methylberberine, berberine acetone, 13-alkylberberine,palmatine, 13-benzylberberine, tetrahydroberberine, tetrahydroprotoberberine, 8-cyanodihydroberberine, 8-oxyberberine, dimeric protoberberine alkaloids,demethylated protoberberine alkaloids, quaternary protoberberine alkaloids,protoberberine and protoberberine alkaloids.

[0150] 4. The composition of embodiment 1, wherein the at least onebioavailability enhancer comprises one or more compound that is capable ofenhancing oral bioavailability and / or efficacy of berberine in the subject.

[0151] 5. The composition of embodiment 1, wherein the at least onebioavailability enhancer comprise one or more compound that is capable of causingdirectly or indirectly one or more of the following in the subject:a) enhancing the solubility or dissolution rate of berberine;b) enhancing the absorption of berberine;c) reducing the clearance of berberine from gastrointestinal tract or blood;d) reducing the rate or extent of metabolism of berberine in liver;e) reducing the rate or extent of distribution of berberine to non-targettissues; andf) enhancing the availability of berberine to the target site of action inother manner.

[0152] 6. The composition of embodiment 1, wherein the at least onebioavailability enhancer comprises a compound that is capable of causing directly orindirectly one or more of the following in the subject:a) inhibiting or reducing P-glycoprotein mediated efflux transport ofberberine; b) inhibiting or reducing the rate or extent of metabolism of berberine orits metabolites in liver;c) inhibiting or reducing the rate or extent of distribution of berberine or itsmetabolites to non-target tissues; andd) inhibiting or reducing cytochrome P450 mediated metabolism ofberberine or its metabolites in gastrointestinal trach and / or liver.Docket Number: 233013-40106

[0153] 7. The composition of embodiment 1, wherein the molar ratio ofberberine and the at least one bioavailability enhancer is about 1:100 to about 100:1.

[0154] 8. The composition of embodiment 1, wherein the at least onebioavailability enhancer comprises a P-glycoprotein inhibitor, a cytochrome P450inhibitor, or a combination thereof.

[0155] 9. The composition of embodiment 8, wherein the P-glycoproteininhibitor is selected from a group consisting of encequidar, phloretin, biochanin A,baicalein, naringenin, apigenin, ketoconazole, KX2-4926, ketoconazole, alkyl(C12−16) dimethylbenzylammonium chloride (Hyamine), cetyltrimethylammoniumbromide (CTAB), magnesium stearate, poly-(ethylene glycol) hexadecyl ether (Brij58), poly(ethylene oxide)20 sorbitan monolaurate (Tween 20), poly(ethylene oxide)20sorbitanmonooleate (Tween 80), poly(ethyleneoxide d-α-tocopheryl polyethyleneglycol 1000 succinate (TPGS),)35 modified castor oil (Cremophor EL), polyethyleneglycol15-hydroxystearate (Solutol HS 15), sodium 1,4-bis (2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate (AOT), sodiumcaprylate (NaCap), sodium deoxycholate(NaDC), sucrose palmitate (Sisterna 16), sucrose stearate (Sisterna 11), hydrolyzedgelatin (HG), hydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC),poly(ethylene oxide)101-block-poly(propylene oxide)56-block-poly(ethylene oxide)101(Pluronic F127), poly(ethylene oxide)80-block-poly(propylene oxide)27-block-poly(ethylene oxide)80 (Pluronic F68), poly(vinyl alcohol) (80% hydrolyzed PVA),poly(vinyl alcohol)-graf t-poly(ethylene glycol) copolymer (Kollicoat), poly-(vinylpyrrolidone) (PVP K30), poly-(vinylpolypyrrolidone) (PVPP), poly(ethyleneglycol) (PEG), and sodium carboxymethylcellulose (NaCMC).

[0156] 10. The composition of embodiment 8, wherein the cytochromeP450 inhibitor is selected from a group consisting of ketoconazole, KX2-4926,Sodium Lauryl Sulfate (SLS), Tween 80, Tween 20, amiodarone, amitriptyline,amlodipine, apigenin, aprepitant, armodafinil, artemisinin, aspirin, barbiturates (e.g.,amobarbital, pentobarbital, phenobarbital, secobarbital), bergamottin (grapefruit),beta-naphthoflavone, bosentan, buprenorphine, bupropion, cafestol, caffeine,cannabis / cannabidiol, carbamazepine, carvedilol, chloramphenicol, chlorphenamine,chlorpromazine, cimetidine, cinacalcet, ciprofloxacin, citalopram, clemastine,clomethiazole, clomipramine, clopidogrel, clofibrate, clotrimazole, cobicistat, codeine,crisaborole, delavirdine, desipramine, dexamethasone, diphenhydramine, diltiazem,Docket Number: 233013-40106dithiocarbamate, domperidone, donepezil, doxepin, doxorubicin, duloxetine,efavirenz, echinacea, enzalutamide, entacapone, escitalopram, erythromycin,ethanolic compounds (e.g., alcohol), esomeprazole, felbamate, fenofibrate,flavonoids (e.g., amentoflavone (Ginkgo biloba, St. John's Wort), grapefruit),fluvoxamine, fluconazole, fluoxetine, fluvastatin, gabapentin, gemfibrozil, gestodene,glutethimide, halofantrine, haloperidol, hyperforin (St. John's Wort), imatinib, indinavir,indomethacin, insulin, interferon, isoniazid, itraconazole, JWH-018, kava,ketoconazole, lansoprazole, letermovir, letrozole, levomepromazine, lovastatin,memantine, metoclopramide, methadone, methimazole, methoxsalen, metyrapone,mibefradil, midodrine, mifepristone, milk thistle, miconazole, moclobemide, modafinil,montelukast, methylcholanthrene, methylphenidate, metronidazole, naringenin(grapefruit), nefazodone, nelfinavir, niacin (nicotinic acid) and niacinamide(nicotinamide), nicotine, nilutamide, norethisterone, norfloxacin, omeprazole,orphenadrine, pantoprazole, paroxetine, perampanel, perphenazine, phenobarbital,phenylbutazone, phenytoin, pilocarpine, piperine, prednisone, pregabalin,probenecid, promethazine, proton pump inhibitors (e.g., esomeprazole, lansoprazole,omeprazole, pantoprazole, rabeprazole), quinidine, quercetin, ranitidine, ribociclib,rifampicin, rifapentine, risperidone, ritonavir, rucaparib, saquinavir, secobarbital,selegiline, seproxetine ((s)-norfluoxetine / fluoxetine metabolite), sertraline, simeprevir,sodium valproate (valproic acid), St. John's Eort, sulconazole, sulfamethoxazole,sulfaphenazole, telithromycin, teniposide, terbinafine, teriflunomide, thiotepa,thiazolidinediones, thioridazine, ticlopidine, tioconazole, tobacco, topiramate,tramadol, tranylcypromine, trimethoprim, trimethoprim / sulfamethoxazole, turmeric,valerian, verapamil, voriconazole, Withania somnifera (ashwagandha), zafirlukast,and zuclopenthixol.

[0157] 11. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises Tween 20 or Tween 80.

[0158] 12. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises Sodium Lauryl Sulfate (SLS).

[0159] 13. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises d-α-tocopheryl polyethylene glycol 1000succinate (TPGS).Docket Number: 233013-40106

[0160] 14. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises Tween 20 and Tween 80.

[0161] 15. The composition of embodiment 8, the at least onebioavailability enhancer comprises Tween 20 and Sodium Lauryl Sulfate (SLS).

[0162] 16. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises Tween 20 and d-α-tocopheryl polyethylene glycol1000 succinate (TPGS).

[0163] 17. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises encequidar.

[0164] 18. The composition of embodiment 8, wherein the at least onebioavailability enhancer comprises encequidar and Tween 20.

[0165] 19. The composition of embodiment 18, wherein the at least onebioavailability enhancer further comprises SLS.

[0166] 20. The composition of any one of the preceding embodiments,further comprising a solid dispersion carrier.

[0167] 21. The composition of embodiment 20, wherein the solid dispersioncarrier comprises mesoporous silicas (e.g., SYLOID® AL-1FP / 63FP silicas,SYLOID® 244 FP silica, SYLOID® XDP silica, and PERKASIL®), and Neusilin.

[0168] 22. The composition of embodiment 11, wherein berberine andTween 20 or berberine and Tween 80 are provided in a molar ratio of about 1:8.

[0169] 23. The composition of embodiment 12, wherein berberine and SLSare provided in a molar ratio of about 1:8.

[0170] 24. The composition of embodiment 14, wherein Tween 20 andTween 80 are present in a ratio of about 1:3.

[0171] 25. The composition of embodiment 14, wherein Tween 20 andTween 80 are present in a ratio of about 3:1.

[0172] 26. The composition of embodiment 15, wherein berberine:Tween20:SLS are provided in a ratio of about 1:4:4.

[0173] 27. The composition of embodiment 16, wherein berberine:Tween20:TPGS are provided in a ratio of about 1:6:2.

[0174] 28. The composition of embodiment 16, wherein berberine:Tween20:TPGS are provided in a ratio of about 1:2:6.Docket Number: 233013-40106

[0175] 29. The composition of embodiment 21, wherein berberine:Tween20:Neusilin are provided in a ratio of 1:8:3.6.

[0176] 30. The composition of embodiment 21, wherein berberine:Tween20:TPGS:Neusilin are provided in a ratio selected from 1:6:2:3.6 or 1:2:6:3.6.

[0177] 31. The composition of embodiment 1, wherein the amount ofberberine is about 0.01% to about 50% by weight.

[0178] 32. The composition of embodiment 1, wherein the composition isformulated in a dosage form selected from the group consisting of a solution,suspensions, tablet, capsule, microemulsion, nano-sized dosage form, liposome,micelle, and phospholipid complex.

[0179] 33. The composition of any one of the preceding embodiments,wherein the composition further comprises a solubilizing agent.

[0180] 34. The composition of embodiment 33, wherein the solubility agentis selected from a group consisting of cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin, (2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD), and sulfobutylether-β-cyclodextrin (SBE-β-CD)), water-soluble organic solvents (e.g. polyethylene glycol300, polyethylene glycol 400, ethanol, propylene glycol, glycerin, N-methyl-2-pyrrolidone, dimethylacetamide, and dimethylsulfoxide), non-ionic surfactants (e.g.,Cremophor EL, Cremophor RH 40, Cremophor RH 60, d-α-tocopherol polyethyleneglycol 1000 succinate, polysorbate 20, polysorbate 80, Solutol HS 15, sorbitanmonooleate, poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol,Gellucire 44 / 14, Softigen 767, and mono- and di-fatty acid esters of PEG 300, 400,or 1750), water-insoluble lipids (e.g.., castor oil, corn oil, cottonseed oil, olive oil,peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenatedvegetable oils, hydrogenated soybean oil, and medium-chain triglycerides of coconutoil and palm seed oil), organic liquids / semi-solids (e.g., beeswax, d-α-tocopherol,oleic acid, medium-chain mono- and diglycerides), and phospholipids (e.g.,hydrogenated soy phosphatidylcholine, distearoylphosphatidylglycerol, l-α-dimyristoylphosphatidylcholine, l-α-dimyristoylphosphatidylglycerol).

[0181] 35. The composition of embodiment 1, wherein oral bioavailability ofberberine in the subject is higher than the oral bioavailability of berberine which isadministered to the subject without the bioavailability enhancer.Docket Number: 233013-40106

[0182] 36. A method of enhancing absorption and / or oral bioavailability ofberberine in a subject by providing to the subject at least one bioavailability enhancerin conjunction with berberine.

[0183] 37. The method of embodiment 36, wherein the at least onebioavailability enhancer comprises a compound that is capable of causing directly orindirectly one or more of the following in the subject:a) enhancing the solubility or dissolution rate of berberine;b) enhancing the absorption of berberine;c) reducing the clearance of berberine from gastrointestinal tract or blood;d) reducing the rate or extent of metabolism of berberine in liver;e) reducing the rate or extent of distribution of berberine to non-targettissues; andf) enhancing the availability of berberine to the target site of action inother manner.

[0184] 38. The method of embodiment 36, wherein the at least onebioavailability enhancer comprises a compound that is capable of causing directly orindirectly one or more of the following in the subject:a) inhibiting or reducing P-glycoprotein mediated efflux transport ofberberine; b) inhibiting or reducing the rate or extent of metabolism of berberine orits metabolites in liver;c) inhibiting or reducing the rate or extent of distribution of berberine or itsmetabolites to non-target tissues; andd) inhibiting or reducing cytochrome P450 mediated metabolism ofberberine or its metabolites in gastrointestinal trach and / or liver.

[0185] 39. The method of embodiment 36-38, wherein the molar ratio ofberberine and the bioavailability enhancer is about 1:100 to about 100:1.

[0186] 40. The method of embodiment 36-38, wherein the at least onebioavailability enhancer comprises a P-glycoprotein inhibitor, a cytochrome P450inhibitor, or a combination thereof.

[0187] 41. The method of embodiment 36-38, wherein the P-glycoproteininhibitor is selected from a group consisting of encequidar, phloretin, biochanin A,baicalein, naringenin, apigenin, ketoconazole, KX2-4926, ketoconazole, alkylDocket Number: 233013-40106(C12−16) dimethylbenzylammonium chloride (Hyamine), cetyltrimethylammoniumbromide (CTAB), magnesium stearate, poly-(ethylene glycol) hexadecyl ether (Brij58), poly(ethylene oxide)20 sorbitan monolaurate (Tween 20), poly(ethylene oxide)20sorbitanmonooleate (Tween 80), poly(ethyleneoxide d-α-tocopheryl polyethyleneglycol 1000 succinate (TPGS),)35 modified castor oil (Cremophor EL), polyethyleneglycol15-hydroxystearate (Solutol HS 15), sodium 1,4-bis (2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate (AOT), sodiumcaprylate (NaCap), sodium deoxycholate(NaDC), sucrose palmitate (Sisterna 16), sucrose stearate (Sisterna 11), hydrolyzedgelatin (HG), hydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC),poly(ethylene oxide)101-block-poly(propylene oxide)56-block-poly(ethylene oxide)101(Pluronic F127), poly(ethylene oxide)80-block-poly(propylene oxide)27-block-poly(ethylene oxide)80 (Pluronic F68), poly(vinyl alcohol) (80% hydrolyzed PVA),poly(vinyl alcohol)-graf t-poly(ethylene glycol) copolymer (Kollicoat), poly-(vinylpyrrolidone) (PVP K30), poly-(vinylpolypyrrolidone) (PVPP), poly(ethyleneglycol) (PEG), and sodium carboxymethylcellulose (NaCMC).

[0188] 42. The method of embodiment 40, wherein the cytochrome P450inhibitor is selected from a group consisting of ketoconazole, KX2-4926, SodiumLauryl Sulfate (SLS), Tween 80, Tween 20, amiodarone, amitriptyline, amlodipine,apigenin, aprepitant, armodafinil, artemisinin, aspirin, barbiturates (e.g., amobarbital,pentobarbital, phenobarbital, secobarbital), bergamottin (grapefruit), beta-naphthoflavone, bosentan, buprenorphine, bupropion, cafestol, caffeine,cannabis / cannabidiol, carbamazepine, carvedilol, chloramphenicol, chlorphenamine,chlorpromazine, cimetidine, cinacalcet, ciprofloxacin, citalopram, clemastine,clomethiazole, clomipramine, clopidogrel, clofibrate, clotrimazole, cobicistat, codeine,crisaborole, delavirdine, desipramine, dexamethasone, diphenhydramine, diltiazem,dithiocarbamate, domperidone, donepezil, doxepin, doxorubicin, duloxetine,efavirenz, echinacea, enzalutamide, entacapone, escitalopram, erythromycin,ethanolic compounds (e.g., alcohol), esomeprazole, felbamate, fenofibrate,flavonoids (e.g., amentoflavone (Ginkgo biloba, St. John's Wort), grapefruit),fluvoxamine, fluconazole, fluoxetine, fluvastatin, gabapentin, gemfibrozil, gestodene,glutethimide, halofantrine, haloperidol, hyperforin (St. John's Wort), imatinib, indinavir,indomethacin, insulin, interferon, isoniazid, itraconazole, JWH-018, kava,ketoconazole, lansoprazole, letermovir, letrozole, levomepromazine, lovastatin,Docket Number: 233013-40106memantine, metoclopramide, methadone, methimazole, methoxsalen, metyrapone,mibefradil, midodrine, mifepristone, milk thistle, miconazole, moclobemide, modafinil,montelukast, methylcholanthrene, methylphenidate, metronidazole, naringenin(grapefruit), nefazodone, nelfinavir, niacin (nicotinic acid) and niacinamide(nicotinamide), nicotine, nilutamide, norethisterone, norfloxacin, omeprazole,orphenadrine, pantoprazole, paroxetine, perampanel, perphenazine, phenobarbital,phenylbutazone, phenytoin, pilocarpine, piperine, prednisone, pregabalin,probenecid, promethazine, proton pump inhibitors (e.g., esomeprazole, lansoprazole,omeprazole, pantoprazole, rabeprazole), quinidine, quercetin, ranitidine, ribociclib,rifampicin, rifapentine, risperidone, ritonavir, rucaparib, saquinavir, secobarbital,selegiline, seproxetine ((s)-norfluoxetine / fluoxetine metabolite), sertraline, simeprevir,sodium valproate (valproic acid), St. John's Eort, sulconazole, sulfamethoxazole,sulfaphenazole, telithromycin, teniposide, terbinafine, teriflunomide, thiotepa,thiazolidinediones, thioridazine, ticlopidine, tioconazole, tobacco, topiramate,tramadol, tranylcypromine, trimethoprim, trimethoprim / sulfamethoxazole, turmeric,valerian, verapamil, voriconazole, Withania somnifera (ashwagandha), zafirlukast,and zuclopenthixol.

[0189] 43. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises Tween 20 or Tween 80.

[0190] 44. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises Sodium Lauryl Sulfate (SLS).

[0191] 45. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises d-α-tocopheryl polyethylene glycol 1000succinate (TPGS).

[0192] 46. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises Tween 20 and Tween 80.

[0193] 47. The method of embodiment 40, the at least one bioavailabilityenhancer comprises Tween 20 and Sodium Lauryl Sulfate (SLS).

[0194] 48. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises Tween 20 and d-α-tocopheryl polyethylene glycol1000 succinate (TPGS).

[0195] 49. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises encequidar.Docket Number: 233013-40106

[0196] 50. The method of embodiment 40, wherein the at least onebioavailability enhancer comprises encequidar and Tween 20.

[0197] 51. The method of embodiment 50, wherein the at least onebioavailability enhancer further comprises SLS.

[0198] 52. The method of any one of embodiments 36-51, wherein a soliddispersion carrier is further administered.

[0199] 53. The method of embodiment 52, wherein the solid dispersioncarrier comprises mesoporous silicas (e.g., SYLOID® AL-1FP / 63FP silicas,SYLOID® 244 FP silica, SYLOID® XDP silica, and PERKASIL®), and Neusilin.

[0200] 54. The method of embodiment 43, wherein berberine and Tween20 or berberine and Tween 80 are provided in a molar ratio of about 1:8.

[0201] 55. The method of embodiment 44, wherein berberine and SLS areprovided in a molar ratio of about 1:8.

[0202] 56. The method of embodiment 46, wherein Tween 20 and Tween80 are present in a ratio of about 1:3.

[0203] 57. The method of embodiment 46, wherein Tween 20 and Tween80 are present in a ratio of about 3:1.

[0204] The method of embodiment 47, wherein berberine:Tween20:SLS are provided in a ratio of about 1:4:4.

[0205] The method of embodiment 48, wherein berberine:Tween20:TPGS are provided in a ratio of about 1:6:2.

[0206] The method of embodiment 48, wherein berberine:Tween20:TPGS are provided in a ratio of about 1:2:6.

[0207] The method of embodiment 53, wherein berberine:Tween20:Neusilin are provided in a ratio of 1:8:3.6.

[0208] The method of embodiment 53, wherein berberine:Tween20:TPGS:Neusilin are provided in a ratio selected from 1:6:2:3.6 or 1:2:6:3.6.

[0209] 63. The method of embodiment 36, wherein the amount of berberineis about 0.01% to about 50% by weight.

[0210] 64. The method of embodiment 36, wherein berberine isadministered to a human subject at a dose ranging from around 100 to about 150 mgper day.Docket Number: 233013-40106

[0211] 67. The method of embodiment 36, wherein berberine isadministered to a human subject at a dose of around 137.4 mg per day.

[0212] 68. The method of embodiment 36, wherein berberine isadministered to a human subject at a dose ranging from around 1,000 mg to 2,000mg per day.

[0213] 69. The method of embodiment 36, wherein berberine isadministered to a human subject at a dose of around 1,714.2 mg per day.

[0214] 70. The method of any one of embodiments 36-69, wherein saidmethod further comprises providing at least one additional bioavailability enhancer tothe subject, prior to the providing to the subject the at least one bioavailabilityenhancer in conjunction with berberine.

[0215] 71. The method of embodiment 70, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one bioavailability enhancerformulated as a single composition. 72. The method of embodiment 70, wherein theproviding to the subject the at least one bioavailability enhancer in conjunction withberberine comprises administrating berberine with the at least one bioavailabilityenhancer formulated as different compositions.

[0216] 73. The method of any one of embodiments 36-69, wherein saidmethod comprises providing a P-glycoprotein inhibitor to the subject prior to theproviding to the subject the at least one bioavailability enhancer in conjunction withberberine.

[0217] 74. The method of embodiment 73, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one bioavailability enhancerformulated as a single composition

[0218] 75. The method of embodiment 73, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one bioavailability enhancerformulated as different compositions.

[0219] 76. The method of any one of embodiments 36-69, said methodcomprises providing a P-glycoprotein inhibitor to the subject prior to the providing tothe subject at least one cytochrome P450 inhibitor.Docket Number: 233013-40106

[0220] 77. The method of embodiment 76, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one cytochrome P450 inhibitorformulated as a single composition.

[0221] 78. The method of embodiment 76, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one cytochrome P450 inhibitorformulated as different compositions.

[0222] 79. The method of any one of embodiments 36-69, said methodcomprises providing encequidar to the subject prior to the providing to the subject atleast one cytochrome P450 inhibitor

[0223] 80. The method of embodiment 79, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one bioavailability enhancerformulated as a single composition.

[0224] 81. The method of embodiment 79, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberinecomprises administrating berberine with the at least one bioavailability enhancerformulated as different compositions.

[0225] 82. The method of any one of embodiment 76-81, wherein thecytochrome P450 inhibitor is Tween 20.

[0226] 83. The method of any one of embodiment 76-81, wherein thecytochrome P450 inhibitor is Sodium Lauryl Sulfate (SLS).

[0227] 84. The method of any one of embodiments 36-69, wherein saidmethod further comprises providing at least one additional bioavailability enhancer tothe subject, prior to the providing to the subject the at least one bioavailabilityenhancer in conjunction with berberine and a solid dispersion carrier.

[0228] 85. The method of embodiment 84, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberine and thesolid dispersion carrier comprises administrating berberine with the at least onebioavailability enhancer and the solid dispersion carrier formulated as a singlecomposition.Docket Number: 233013-40106

[0229] 86. The method of embodiment 84, wherein the providing to thesubject the at least one bioavailability enhancer in conjunction with berberine and thesolid dispersion carrier comprises administrating berberine with the at least onebioavailability enhancer and the solid dispersion carrier formulated as differentcompositions.

[0230] 87. The method of any of one of embodiments 84-86, wherein the atleast one additional bioavailability enhancer comprises encequidar, the at least onebioavailability enhancer comprises Tween 20, and the solid dispersion carriercomprises Neusilin.

[0231] 88. The methods of any one of embodiments 36-87, wherein asolubilizing agent is further administered.

[0232] 89. The method of embodiment 90, wherein the solubilizing agent isselected from a group consisting of cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin,(2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD), and sulfobutylether-β-cyclodextrin(SBE-β-CD)), water-soluble organic solvents (e.g. polyethylene glycol 300,polyethylene glycol 400, ethanol, propylene glycol, glycerin, N-methyl-2-pyrrolidone,dimethylacetamide, and dimethylsulfoxide), non-ionic surfactants (e.g., CremophorEL, Cremophor RH 40, Cremophor RH 60, d-α-tocopherol polyethylene glycol 1000succinate, polysorbate 20, polysorbate 80, Solutol HS 15, sorbitan monooleate,poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol, Gellucire 44 / 14,Softigen 767, and mono- and di-fatty acid esters of PEG 300, 400, or 1750), water-insoluble lipids (e.g.., castor oil, corn oil, cottonseed oil, olive oil, peanut oil,peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils,hydrogenated soybean oil, and medium-chain triglycerides of coconut oil and palmseed oil), organic liquids / semi-solids (e.g., beeswax, d-α-tocopherol, oleic acid,medium-chain mono- and diglycerides), and phospholipids (e.g., hydrogenated soyphosphatidylcholine, distearoylphosphatidylglycerol, l-α-dimyristoylphosphatidylcholine, l-α-dimyristoylphosphatidylglycerol).

[0233] 90. The method of embodiment 36, wherein said method comprisesproviding at least one additional bioavailability enhancer to the subject prior to theadministration of the composition of any one of embodiments 1-35.Docket Number: 233013-40106

[0234] 91. The method of embodiment 35, wherein said method comprisesproviding encequidar to the subject prior to the administration of the composition ofany one of embodiments 1-35.

[0235] 92. A method of treating, preventing and / or modulating a disease ora condition in a subject comprising administering to the subject an effective amountof berberine and a bioavailability enhancer.

[0236] 93. A method of treating, preventing and / or modulating a disease ora condition in a subject comprising administering to the subject the composition ofany one of embodiments 1-35.

[0237] 94. The method of embodiment 92 or 93, wherein the administrationis performed between one and three times per day.

[0238] 95. The method of embodiment 92 or 93, wherein the disease orcondition is selected from the group consisting of diabetes, dyslipidemia, metabolicdisorders, cardiovascular diseases, hyperlipidemia, fatty liver, inflammation,Alzheimer’s disease, lung diseases and infectious diseases.

[0239] 96. The method of embodiment 92 or 93, wherein the absorption ororal bioavailability of berberine in the subject is higher than the absorption or oralbioavailability of berberine which is administered to the subject without thebioavailability enhancer.

[0240] 97. A method of modulating an activity or function of a moleculartarget in a subject comprising administering to the subject berberine and abioavailability enhancer.

[0241] 98. A method of modulating an activity or function of a moleculartarget in a subject comprising administering to the subject the composition of anyone of embodiments 1-35.

[0242] 99. The method of embodiment 97 or 98, wherein the moleculartarget is selected from the group consisting of AMPK, CD36, acetyl-CoA carboxylase(ACC), microsomal triglyceride transfer protein (MTTP), scavenger receptor class Btype 1 (SR-BI), low-density lipoprotein receptor (LDLR), ATP-binding cassettetransporter A1 (ABCA1), SIRT-, endothelial nitric oxide synthetase (eNOS), lectin-like low-density lipoprotein receptor-1 (LOX-1), PI3K / AKT (PKB), mTOR, JAK2,calcium channels, NFkB, a target gene of endoplasmic reticulum stress, and HIF-1α.

Claims

Docket Number: 233013-40106CLAIMSWhat is claimed is:

1. A composition for the treatment, prevention and / or modulation of a disease ora conditions in a subject, comprising a therapeutically effective amount of berberineand at least one bioavailability enhancer, or their respective pharmaceuticallyacceptable derivatives selected from the group consisting of salts, esters, enolethers, enol esters, acetals, ketals, orthoesters, hemiacetals, hemiketals, acids,bases, and hydrates thereof.

2. The composition of claim 1, wherein the berberine is present as a saltselected from the group consisting of berberine hydrochloride, berberine chloride,berberine sulfate, berberine phosphate, berberine acetate, berberine succinate,berberine bromide, berberine iodide, berberine citrate, berberine lactate, berberinemalate, berberine glutamate, and berberine tannate.

3. The composition of claim 1, wherein the berberine is present as a derivativeselected from the group consisting of berberine alkaloid, berberine base, berberinehydrochloride, berberine, berberrubine, coreximine, tetrahydropalmatine, jatrorrhizine,13-hydroxyberberine chloride, coralyne, coralyne chloride, 7,8-dihydro-13-methylberberine, berberine acetone, 13-alkylberberine, palmatine, 13-benzylberberine, tetrahydroberberine, tetrahydroprotoberberine, 8-cyanodihydroberberine, 8-oxyberberine, dimeric protoberberine alkaloids,demethylated protoberberine alkaloids, quaternary protoberberine alkaloids,protoberberine and protoberberine alkaloids.

4. The composition of claim 1, wherein the molar ratio of berberine and the atleast one bioavailability enhancer is about 1:100 to about 100:1.

5. The composition of claim 1, wherein the at least one bioavailability enhancercomprises a P-glycoprotein inhibitor, a cytochrome P450 inhibitor, or a combinationthereof.Docket Number: 233013-401066. The composition of claim 5, wherein the P-glycoprotein inhibitor is selectedfrom a group consisting of encequidar, phloretin, biochanin A, baicalein, naringenin,apigenin, ketoconazole, KX2-4926, ketoconazole, alkyl (C12−16)dimethylbenzylammonium chloride (Hyamine), cetyltrimethylammonium bromide(CTAB), magnesium stearate, poly-(ethylene glycol) hexadecyl ether (Brij 58),poly(ethylene oxide)20 sorbitan monolaurate (Tween 20), poly(ethylene oxide)20sorbitanmonooleate (Tween 80), poly(ethyleneoxide d-α-tocopheryl polyethyleneglycol 1000 succinate (TPGS),)35 modified castor oil (Cremophor EL), polyethyleneglycol15-hydroxystearate (Solutol HS 15), sodium 1,4-bis (2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate (AOT), sodiumcaprylate (NaCap), sodium deoxycholate(NaDC), sucrose palmitate (Sisterna 16), sucrose stearate (Sisterna 11), hydrolyzedgelatin (HG), hydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC),poly(ethylene oxide)101-block-poly(propylene oxide)56-block-poly(ethylene oxide)101(Pluronic F127), poly(ethylene oxide)80-block-poly(propylene oxide)27-block-poly(ethylene oxide)80 (Pluronic F68), poly(vinyl alcohol) (80% hydrolyzed PVA),poly(vinyl alcohol)-graf t-poly(ethylene glycol) copolymer (Kollicoat), poly-(vinylpyrrolidone) (PVP K30), poly-(vinylpolypyrrolidone) (PVPP), poly(ethyleneglycol) (PEG), and sodium carboxymethylcellulose (NaCMC).

7. The composition of claim 5, wherein the cytochrome P450 inhibitor is selectedfrom a group consisting of ketoconazole, KX2-4926, Sodium Lauryl Sulfate (SLS),Tween 80, Tween 20, amiodarone, amitriptyline, amlodipine, apigenin, aprepitant,armodafinil, artemisinin, aspirin, barbiturates (e.g., amobarbital, pentobarbital,phenobarbital, secobarbital), bergamottin (grapefruit), beta-naphthoflavone,bosentan, buprenorphine, bupropion, cafestol, caffeine, cannabis / cannabidiol,carbamazepine, carvedilol, chloramphenicol, chlorphenamine, chlorpromazine,cimetidine, cinacalcet, ciprofloxacin, citalopram, clemastine, clomethiazole,clomipramine, clopidogrel, clofibrate, clotrimazole, cobicistat, codeine, crisaborole,delavirdine, desipramine, dexamethasone, diphenhydramine, diltiazem,dithiocarbamate, domperidone, donepezil, doxepin, doxorubicin, duloxetine,efavirenz, echinacea, enzalutamide, entacapone, escitalopram, erythromycin,ethanolic compounds (e.g., alcohol), esomeprazole, felbamate, fenofibrate,flavonoids (e.g., amentoflavone (Ginkgo biloba, St. John's Wort), grapefruit),fluvoxamine, fluconazole, fluoxetine, fluvastatin, gabapentin, gemfibrozil, gestodene,Docket Number: 233013-40106glutethimide, halofantrine, haloperidol, hyperforin (St. John's Wort), imatinib, indinavir,indomethacin, insulin, interferon, isoniazid, itraconazole, JWH-018, kava,ketoconazole, lansoprazole, letermovir, letrozole, levomepromazine, lovastatin,memantine, metoclopramide, methadone, methimazole, methoxsalen, metyrapone,mibefradil, midodrine, mifepristone, milk thistle, miconazole, moclobemide, modafinil,montelukast, methylcholanthrene, methylphenidate, metronidazole, naringenin(grapefruit), nefazodone, nelfinavir, niacin (nicotinic acid) and niacinamide(nicotinamide), nicotine, nilutamide, norethisterone, norfloxacin, omeprazole,orphenadrine, pantoprazole, paroxetine, perampanel, perphenazine, phenobarbital,phenylbutazone, phenytoin, pilocarpine, piperine, prednisone, pregabalin,probenecid, promethazine, proton pump inhibitors (e.g., esomeprazole, lansoprazole,omeprazole, pantoprazole, rabeprazole), quinidine, quercetin, ranitidine, ribociclib,rifampicin, rifapentine, risperidone, ritonavir, rucaparib, saquinavir, secobarbital,selegiline, seproxetine ((s)-norfluoxetine / fluoxetine metabolite), sertraline, simeprevir,sodium valproate (valproic acid), St. John's Eort, sulconazole, sulfamethoxazole,sulfaphenazole, telithromycin, teniposide, terbinafine, teriflunomide, thiotepa,thiazolidinediones, thioridazine, ticlopidine, tioconazole, tobacco, topiramate,tramadol, tranylcypromine, trimethoprim, trimethoprim / sulfamethoxazole, turmeric,valerian, verapamil, voriconazole, Withania somnifera (ashwagandha), zafirlukast,and zuclopenthixol.

8. The composition of claim 1, wherein the composition further comprises asolubilizing agent.

9. The composition of claim 1, wherein the solubility agent is selected from agroup consisting of cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin, (2-Hydroxypropyl)-β-cyclodextrin (HP-β-CD), and sulfobutylether-β-cyclodextrin (SBE-β-CD)), water-soluble organic solvents (e.g. polyethylene glycol 300, polyethyleneglycol 400, ethanol, propylene glycol, glycerin, N-methyl-2-pyrrolidone,dimethylacetamide, and dimethylsulfoxide), non-ionic surfactants (e.g., CremophorEL, Cremophor RH 40, Cremophor RH 60, d-α-tocopherol polyethylene glycol 1000succinate, polysorbate 20, polysorbate 80, Solutol HS 15, sorbitan monooleate,poloxamer 407, Labrafil M-1944CS, Labrafil M-2125CS, Labrasol, Gellucire 44 / 14,Softigen 767, and mono- and di-fatty acid esters of PEG 300, 400, or 1750), water-Docket Number: 233013-40106insoluble lipids (e.g., castor oil, corn oil, cottonseed oil, olive oil, peanut oil,peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils,hydrogenated soybean oil, and medium-chain triglycerides of coconut oil and palmseed oil), organic liquids / semi-solids (e.g., beeswax, d-α-tocopherol, oleic acid,medium-chain mono- and diglycerides), and phospholipids (e.g., hydrogenated soyphosphatidylcholine, distearoylphosphatidylglycerol, l-α-dimyristoylphosphatidylcholine, l-α-dimyristoylphosphatidylglycerol).

10. A method of enhancing absorption and / or oral bioavailability of berberine in asubject by providing to the subject the composition of claims 1-9.