Patent Publication Number: US-2016220524-A1

Title: Compositions of small molecule therapeutics

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application claims the priority of U.S. Provisional Application Ser. No. 61/343,781, filed May 4, 2010, the disclosures of which are incorporated in their entirety herein by reference. 
    
    
     FIELD OF INVENTION 
     This invention relates to compositions of small molecule therapeutics with enhanced pharmacokinetic properties. 
     BACKGROUND OF INVENTION 
     Small molecule drug discovery is actively pursued by biotech companies to complement the therapeutic advances made with protein based drugs such as recombinant proteins and monoclonal antibodies. Moreover, small molecule drug treatments often may enjoy cost benefits. Patients are also more likely to accept orally available small molecule treatments than the typical injectable protein based drug. It has now been found that the pharmacokinetic properties of small molecule therapeutics can be improved by the co-administration of certain enhancers. 
     SUMMARY OF THE INVENTION 
     Pharmacokinetic properties of small molecule therapeutics such as the taxanes and small molecule drug substances classified according to the Biopharmaceutics Classification System (BCS) as Class 2-4 compounds are enhanced by the addition of an alkyl N,N-disubstituted-amino acetate in free base or salt form. A greater systemic exposure, higher peak plasma levels, and longer mean residence time can be achieved. 
     Particularly preferred are compositions comprising paclitaxel and dodecyl 2-(N,N-dimethylamino) propionate (DDAIP) in, free base or salt form. 
     The solubility and absorption of small molecule drug substances classified as BCS 2 and BCS 4 compounds, are particularly enhanced by dodecyl 2-(N,N-dimethylamino) propionate in free base or salt form. Particularly preferred are the BCS Class 2 compounds, lansoprazole, haloperidol and sulfasalazine, the BCS Class 3 compounds, atenolol and glucosamine; and the BCS Class 4 compounds, furosemide and chlorothiazide. 
     Preferred routes of administration are oral and subcutaneous. 
    
    
     
       BRIEF DESCRIPTION OF DRAWINGS 
         FIG. 1  is a graphical representation of paclitaxel concentrations in rat plasma samples after oral (PO) dosing (5 mg/kg); data points represent mean values and error bars represent standard errors of the mean values. 
         FIG. 2  is a graphical representation of paclitaxel concentrations in rat plasma samples after subcutaneous (SC) dosing (5 mg/kg); data points represent mean values and error bars represent standard errors of the mean values. 
         FIG. 3  is another graphical representation of paclitaxel concentrations in rat plasma samples after oral (PO) dosing (5 mg/kg); data points represent mean values and error bars represent standard errors of the mean values. 
         FIG. 4  is a graphical representation of lansoprazole and DDAIP effects on gastric emptying in mice. 
         FIG. 5  is a graphical representation of pharmacokinetic profile of lansoprazole in dog plasma samples after oral administration with and without DDAIP. 
     
    
    
     DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS 
     As used herein, the term “small molecule therapeutic” denotes a low molecular weight organic compound which is not a polymer but binds with relatively high affinity to a biopolymer such as a protein, a nucleic acid, or polysaccharide and also alters the activity or function of the biopolymer. The upper molecular weight limit for a small molecule therapeutic is about 1000 Daltons which allows for diffusion across all membranes so that intracellular sites of action can be reached. Very small oligomers are also considered small molecules, e.g., dinucleotides, disaccharides, and the like. Illustrative are paclitaxel, DHA-paclitaxel, mesalamine (Pentasa®), motexafin gadolinium, temozolomide, erlotinib (Tarceva®, cinacalcet (Sensipar®), safinamide, simvastatin (Zocor®), pravastatin (Pravachol®), sildenafil, peptide mimetics, the siRNAs, and the like. 
     Taxanes are diterpenes utilized in cancer chemotherapy. Particularly well suited for purposes of the present invention are paclitaxel, docetaxel, tesetaxel, and mixture thereof. 
     Also suited for purposes of the present invention are small molecule therapeutic compounds that are potent and pharmaceutically relevant but usually poorly soluble or insoluble compounds. A Biopharmaceutics Classification System (BCS) is a guide provided by the U.S. Food and Drug Administration for predicting the intestinal drug absorption. The system correlates in vitro drug product dissolution and in vivo bioavailability based on recognition that drug dissolution and gastrointestinal permeability are fundamental parameters controlling rate and extent of drug absorption. Four drug classes are defined in an article by Amidon G. L, et al., “ A theoretical basis for a biopharmaceutic drug classification: the correlation of in vitro product dissolution and in vivo bioavailability.” Pharm. Res.,  12(3), 413-20 (1995). 
     According to the Biopharmaceutics Classification System, drug substances are classified as follows: 
     Class 1—High permeability, High solubility drugs. Those compounds are well absorbed and their absorption rate is usually higher than excretion. An example compound is metoprolol. 
     Class 2—High permeability, Low solubility drugs. The bioavailability of those products is limited by their solvation rate. A correlation between the in vivo bioavailability and the in vitro solvation can be found. Example compounds are lansoprazole, haloperidol, sulfasalazine, and glibenclamide. 
     Class 3—Low permeability, High solubility drugs. The absorption is limited by the permeation rate but the drug is solvated very fast. Example compounds are atenolol, glucosamine (or salts thereof), and cimetidine. 
     Class 4—Low permeability, Low solubility drugs. Those compounds have a poor bioavailability. Usually they are not well absorbed over the intestinal mucosa and a high variability is expected. Example compounds are furosemide, chlorothiazide, and hydrochlorothiazide. 
     The drugs are classified in BCS on the basis of the following parameters: 1. solubility; 2. permeability; and 3. dissolution. The class boundaries for solubility are based on the highest dose strength of an immediate release product. A drug is considered highly soluble when the highest dose strength is soluble in 250 milliliters (ml) or less of aqueous media over the pH range of 1 to 7.5. The class boundaries for permeability are based indirectly on the extent of absorption of a drug substance in humans and directly on the measurement of rates of mass transfer across human intestinal membrane. Alternative non-human systems capable of predicting the drug absorption in humans can be used (such as in vitro culture methods). A drug substance is considered highly permeable when the extent of absorption in humans is determined to be 90% or more of the administered dose based on a mass-balance determination or in comparison to an intravenous dose. The class boundaries for dissolution for an immediate release product to be considered rapidly dissolving is when no less than 85% of the labeled amount of the drug substance dissolves within 30 minutes using a USP Dissolution Apparatus 1 at 100 RPM or Apparatus 2 at 50 RPM in a volume of 900 ml or less in a media of 0.1N HCl or simulated gastric fluid or pH 4.5 buffer and pH 6.8 buffer or simulated intestinal fluid. 
     Class 2-4 compounds are particularly well suited for compositions embodying this invention. Preferred are small molecule therapeutic compounds appearing in the  WHO List of Essential Drugs,  16 th  Ed., revised March 2010. Particularly preferred small molecule therapeutic compounds are lansoprazole, haloperidol, and sulfasalazine in Class 2; atenolol, and glucosamine in Class 3; and furosemide and chlorothiazide in Class 4. 
     The alkyl N,N-disubstituted amino acetates suitable for purposes of the present invention are represented by the formula: 
     
       
         
         
             
             
         
       
     
     wherein n is an integer having a value in the range of about 4 to about 18; R is a member of the group consisting of hydrogen, C 1  to C 7  alkyl, benzyl and phenyl; R 1  and R 2  are members of the group consisting of hydrogen and C 1  to C 7  alkyl; R 3  and R 4  are members of the group consisting of hydrogen, methyl and ethyl. 
     Preferred alkyl (N,N-disubstituted amino)-acetates are C 4  to C 18  alkyl (N,N-disubstituted amino)-acetates and C 4  to C 18  alkyl (N,N-disubstituted amino)-propionates as well as pharmaceutically acceptable salts and derivatives thereof. Exemplary specific alkyl-2-(N,N-disubstituted amino)-acetates include 
     
       
         
         
             
             
         
       
     
     dodecyl 2-(dimethylamino)-propionate (DDAIP) and 
     
       
         
         
             
             
         
       
     
     dodecyl 2-(N,N-dimethylamino)-acetate (DDAA) 
     Alkyl-2-(N,N-disubstituted amino) acetates are known. For example, dodecyl 2-(N,N-dimethylamino)-propionate (DDAIP) is available from Steroids, Ltd. (Chicago, Ill.). In addition, alkyl-2-(N,N-disubstituted amino) alkanoates can be synthesized from more readily available compounds as described in U.S. Pat. No. 4,980,378 to Wong et al., which is incorporated herein by reference to the extent that it is not inconsistent. As described therein, alkyl-2-(N,N-disubstituted amino) acetates are readily prepared via a two-step synthesis. In the first step, long chain alkyl chloroacetates are prepared by reaction of the corresponding long chain alkanols with chloromethyl chloroformate or the like in the presence of an appropriate base such as triethylamine, typically in suitable solvent such as chloroform. The reaction can be depicted as follows: 
     
       
         
         
             
             
         
       
     
     wherein n, R, R 1 , R 2 , R 3  and R 4  are defined as above. The reaction temperature may be selected from about 10 degrees Celsius to about 200 degrees Celsius or reflux, with room temperature being preferred. The use of a solvent is optional. If a solvent is used, a wide variety of organic solvents may be selected. Choice of a base is likewise not critical. Preferred bases include tertiary amines such as triethylamine, pyridine and the like. Reaction time generally extends from about one hour to three days. 
     In the second step, the long chain alkyl haloacetate such as chloroacetate is condensed with an appropriate amine according to the scheme: 
     
       
         
         
             
             
         
       
     
     wherein n, R, R 1 , R 2 , R 3  and R 4  are defined as before. Excess amine reactant is typically used as the base and the reaction is conveniently conducted in a suitable solvent such as ether. This second step is preferably run at room temperature, although temperature may vary. Reaction time usually varies from about one hour to several days. Conventional purification techniques can be applied to ready the resulting ester for use in a pharmaceutical composition. 
     The free base forms of the foregoing compounds are liquids at ambient temperature. The salt forms, on the other hand, are solids. For administration to a patient, both forms can be combined with the small molecule therapeutic in a physiologically acceptable carrier such as water or water-solvent admixture depending upon the solubility of the small molecule therapeutic. A pharmacologically acceptable carrier for the active ingredient and the alkyl N,N-disubstituted amino acetate can be a liquid or a solid. The choice of the particular carrier is dictated usually by the active ingredient. The term “pharmaceutically acceptable carrier” is art-recognized and refers to a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the active ingredient from one, organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the active ingredient and not injurious to the patient. Some examples of materials which may serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pryogen-free water; (17) isotonic saline; (18) Ringer&#39;s solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. 
     An illustrative water-solvent carrier for taxanes that is physiologically acceptable is a water-polyethylene glycol (PEG) admixture containing about 10 to about 20 percent (v/v) polyethylene glycol 300 (PEG 300) or polyethylene glycol 400 (PEG 400). 
     The amount of taxane administered to a patient can vary, but usually is in the range of about 200 to 250 mg/m 2 . In one embodiment, paclitaxel (Taxol®) was formulated in either 10% polyethylene glycol 400 (PEG 400) in water, or in aqueous solutions of DDAIP, or its hydrochloric salt, DDAIP*HCL (40%). 
     Example I 
     Using Sprague Dawley rats as an animal model, the body weights of the animals were measured prior to dosing to determine the appropriate amount of test article to deliver. Animals were dosed (5 mg/kg) by oral gavage (PO) or subcutaneously (SC) with the test composition in the different formulations as a single bolus. Then blood samples were taken via the lateral tail vein at 0.5, 1, 2, 4, 8 and 24 hours following the dose. 
     Blood samples were collected into blood collection tubes containing Na 2 EDTA, placed on ice and within 30 minutes of sampling the blood samples were centrifuged to obtain plasma. The plasma was separated from the cellular component and placed in microcentrifuge tubes and frozen, then stored at −80° C. until processed for analysis by LC-MS/MS (liquid chromatography-mass spectrometry with peptide mass fingerprinting). Standard samples were created by adding known amounts of paclitaxel to blank rat plasma. Standard curves were created by analyzing the standard samples by LC-MS/MS and using the resultant areas under the chromatographic peaks in conjunction with the known concentration of the samples. 
     The experimental samples were analyzed using the same method and the areas under the chromatographic peaks were used in conjunction with the standard curves to calculate the concentration of paclitaxel in the samples. The peak plasma concentrations (C max ) of paclitaxel were increased 10-fold after oral dosing and 2-fold after subcutaneous dosing in DDAIP-HCL formulation as compared to the PEG formulation. Use of the DDAIP-HCL formulation resulted in longer absorption phases and later onset of T max  times when dosed both orally and subcutaneously compared to the use of the PEG formulation. 
     The overall systemic exposure (AUC) and bioequivalence were increased with the DDAIP*HCL formulation as compared to when the PEG 400 formulation was used. After oral administration with the DDAIP-HCL formulation, the overall systemic exposure of paclitaxel was increased approximately 75 times and after subcutaneous dosing approximately 20 times compared to the PEG formulations via the same administration routes. The mean residence time (MRT) increased to 9.8 hours after oral dosing and 11.5 hours after subcutaneous dosing when the DDAIP*HCL formulation was used compared to the PEG 400 formulation (3.8 hours PO and 8.5 hours SC). Additionally, dosing with DDAIP-HCL, the hydrochloride salt of DDAIP, resulted in greater systemic exposure, higher peak plasma levels and longer MRT compared to DDAIP alone. The experimental results are graphically presented in  FIGS. 1 and 2 . 
     It can be concluded from these results that the formulation DDAIP.HCL provides superior pharmacokinetic properties to paclitaxel compared to the polyethylene glycol (PEG 400) formulation. Additionally, the DDAIP-HCL formulation provided superior pharmacokinetic properties to paclitaxel compared to DDAIP alone. 
     Standard samples for PO delivery of paclitaxel (Taxol®) formulated in 10% PEG 400, DDAIP and 40% DDAIP.HCL were created by adding known amounts of paclitaxel to blank rat plasma. The Standard samples were analyzed by LC-MC/MS and the resultant areas under the chromatographic peaks were used to construct a standard curve. The results are shown in Tables I-III for the PO delivery. 
     
       
         
           
               
             
               
                 TABLE I 
               
             
            
               
                   
               
               
                 Results of the LC-MS/MS Analysis of the Standard Samples. 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Mean 
                   
                   
                 Mean 
                   
               
               
                 Actual 
                 Area 
                 First 
                 Second 
                 Measured 
                   
               
               
                 Conc. 
                 Ratio 
                 Injection 
                 Injection 
                 Conc. 
                 Accuracy 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 0.5 
                 0.00365 
                 0.55 
                 0.448 
                 0.449 
                 99.80 
               
               
                 1 
                 0.0061 
                 1.045 
                 1.274 
                 1.1595 
                 115.95 
               
               
                 5 
                 0.0215 
                 4.475 
                 4.565 
                 4.52 
                 90.40 
               
               
                 10 
                 0.0454 
                 9.915 
                 9.876 
                 9.8955 
                 98.96 
               
               
                 25 
                 0.1144 
                 24.346 
                 26.561 
                 25.4535 
                 101.81 
               
               
                 50 
                 0.2218 
                 47.726 
                 51.603 
                 49.6645 
                 99.33 
               
               
                 75 
                 0.33305 
                 70.706 
                 78.75 
                 74.728 
                 99.64 
               
               
                 100 
                 0.4797 
                 102.494 
                 113.091 
                 107.7925 
                 107.79 
               
               
                   
               
            
           
         
       
     
     The best fit straight line by least squares linear regression is represented by the formula: 
       Area Ratio=4.66 E -3 (conc&#39;n in ng/ml)−2.19 E -03 R   2 −9.97 E -01
 
     
       
         
           
               
             
               
                 TABLE II 
               
             
            
               
                   
               
               
                 Results of the LC-MS/MS Analysis of the PO Samples. 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 Formula 
                 Time (Hr.) 
                 R-1 
                 R-2 
                 Mean 
                 SEM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 10% 
                 0 
                 0.232 
                 0 
                 0.1 
                 0.1 
               
               
                   
                 PEG400 
                 0.5 
                 0.325 
                 0.351 
                 0.3 
                 0.0 
               
               
                   
                   
                 1 
                 0.48 
                 0 
                 0.0 
                 0.0 
               
               
                   
                   
                 2 
                 0 
                 1.137 
                 0.8 
                 0.3 
               
               
                   
                   
                 4 
                 0 
                 0.369 
                 0.2 
                 0.2 
               
               
                   
                   
                 8 
                 0 
                 0.481 
                 0.2 
                 0.2 
               
               
                   
                   
                 24 
                 0 
                 0 
                 0.0 
                 0.0 
               
               
                   
                 DDAIP 
                 0 
                 0 
                 0 
                 0.0 
                 0.0 
               
               
                   
                   
                 0.5 
                 2.963 
                 3.215 
                 3.1 
                 0.1 
               
               
                   
                   
                 1 
                 3.32 
                 3.092 
                 3.2 
                 0.1 
               
               
                   
                   
                 2 
                 4.539 
                 5.306 
                 4.9 
                 0.4 
               
               
                   
                   
                 4 
                 2.679 
                 2.376 
                 2.5 
                 0.2 
               
               
                   
                   
                 8 
                 1.459 
                 1.331 
                 1.4 
                 0.1 
               
               
                   
                   
                 24 
                 0.487 
                 0.556 
                 0.5 
                 0.0 
               
               
                   
                 40% 
                 0 
                 0 
                 0 
                 0.0 
                 0.0 
               
               
                   
                 DDAIP 
                 0.5 
                 1.401 
                 1.159 
                 1.3 
                 0.1 
               
               
                   
                 HCl 
                 1 
                 4.168 
                 3.652 
                 3.9 
                 0.3 
               
               
                   
                   
                 2 
                 5.169 
                 4.872 
                 5.0 
                 0.1 
               
               
                   
                   
                 4 
                 9.699 
                 8.008 
                 8.9 
                 0.8 
               
               
                   
                   
                 8 
                 6.009 
                 5.124 
                 5.6 
                 0.4 
               
               
                   
                   
                 24 
                 3.012 
                 3.057 
                 3.0 
                 0.0 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE III 
               
             
            
               
                   
               
               
                 Derivation of Pharmacokinetic Parameters from the PO 
               
               
                 Data. 
               
            
           
           
               
               
               
               
               
               
            
               
                 Formulation 
                 Parameter (units) 
                 R-1 
                 R-2 
                 Mean 
                 SEM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 10% PEG400 
                 C max  (ng/mL) 
                 0.5 
                 1.1 
                 0.8 
                 0.46 
               
               
                   
                 T max  (hr.) 
                 2 
                 2 
                 2.0 
                 0.00 
               
               
                   
                 AUC (ng * hr/mL) 
                 0.9 
                 7.8 
                 4.4 
                 4.85 
               
               
                   
                 Mean Residence Time 
                 1.6 
                 6.0 
                 3.8 
                 3.06 
               
               
                   
                 (MRT, hr.) 
                   
                   
                   
                   
               
               
                   
                 T½ MRT  (hr.) 
                 1.1 
                 4.1 
                 2.6 
                 2.12 
               
               
                 DDAIP 
                 C max  (ng/mL) 
                 4.5 
                 5.3 
                 4.9 
                 0.54 
               
               
                   
                 T max  (hr.) 
                 2 
                 2 
                 2.0 
                 0.00 
               
               
                   
                 AUC (ng * hr/mL) 
                 37.3 
                 36.8 
                 37.0 
                 0.38 
               
               
                   
                 Mean Residence Time 
                 6.9 
                 7.1 
                 7.0 
                 0.10 
               
               
                   
                 (MRT, hr.) 
                   
                   
                   
                   
               
               
                   
                 T½ MRT  (hr.) 
                 4.8 
                 4.9 
                 4.9 
                 0.07 
               
               
                 40% DDAIP 
                 C max  (ng/mL) 
                 9.7 
                 8.0 
                 8.9 
                 1.20 
               
               
                 HCl 
                 T max  (hr.) 
                 4 
                 4 
                 4.0 
                 0.00 
               
               
                   
                 AUC (ng * hr/mL) 
                 124.9 
                 110.3 
                 117.6 
                 10.26 
               
               
                   
                 Mean Residence Time 
                 9.6 
                 10.1 
                 9.8 
                 0.35 
               
               
                   
                 (MRT, hr.) 
                   
                   
                   
                   
               
               
                   
                 T½ MRT  (hr.) 
                 6.6 
                 7.0 
                 6.8 
                 0.24 
               
            
           
           
               
               
               
               
               
            
               
                 Bioequivalence (AUC DDAIP / 
                 39.7 
                 4.7 
                 22.2 
                 24.71 
               
               
                 AUC 10% PEG ) 
                   
                   
                   
                   
               
               
                 Bioequivalence (AUC 40% DDAIP•HCL / 
                 132.8 
                 41.2 
                 73.5 
                 83.87 
               
               
                 AUC 10% PEG ) 
               
               
                   
               
               
                 T½MRT = 0.693 * MRT 
               
            
           
         
       
     
     Standard samples for SC delivery of paclitaxel formulated in 10% PEG 400 and 40% DDAIP*HCL were created by adding known amounts of paclitaxel to blank rat plasma. The Standard samples were analyzed by LC-MS/MS, and the resultant areas under the chromatographic peaks were used to construct a standard curve. and in Table IV and VI for the SC delivery. 
     
       
         
           
               
             
               
                 TABLE IV 
               
             
            
               
                   
               
               
                 Results of the LC-MS/MS analysis of the Standard Samples. 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Mean 
                   
                   
                 Mean 
                   
               
               
                 Actual 
                 Area 
                 First 
                 Second 
                 Measured 
                   
               
               
                 Conc. 
                 Ratio 
                 Injection 
                 Injection 
                 Conc. 
                 Accuracy 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 0.2 
                 0.0008 
                 N/A 
                 0.211 
                 0.211 
                 105.50 
               
               
                 0.5 
                 0.00135 
                 0.426 
                 0.463 
                 0.4445 
                 88.90 
               
               
                 1 
                 0.00285 
                 1.063 
                 1.114 
                 1.0885 
                 108.85 
               
               
                 5 
                 0.01185 
                 4.876 
                 5.066 
                 4.971 
                 99.42 
               
               
                 10 
                 0.02335 
                 9.705 
                 10.103 
                 9.904 
                 99.04 
               
               
                 25 
                 0.0617 
                 26.224 
                 26.661 
                 26.4425 
                 105.77 
               
               
                 50 
                 0.1092 
                 46.623 
                 47.127 
                 46.875 
                 93.75 
               
               
                 75 
                 0.1765 
                 75.029 
                 76.672 
                 75.8505 
                 101.13 
               
               
                 100 
                 0.2335 
                 101.111 
                 99.734 
                 100.4225 
                 100.42 
               
               
                   
               
            
           
         
       
     
     The best fit straight line by least squares linear regression is represented by the formula: 
       Area Ratio=2.32 E -03 (conc&#39;n in ng/ml)+1.98 E -04  R   2 =9.99 E -01 
     
       
         
           
               
             
               
                 TABLE V 
               
             
            
               
                   
               
               
                 Results of the LC-MS/MS Analysis of the Experimental SC 
               
               
                 Samples. 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Formula 
                 Time (Hr.) 
                 R-1 
                 R-2 
                 R-3 
                 Mean 
                 SEM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 10% PEG 400 
                 0 
                 0 
                 0 
                 0 
                 0.0 
                 0.0 
               
               
                   
                 0.5 
                 10.913 
                 12.249 
                 12.774 
                 11.6 
                 0.7 
               
               
                   
                 1 
                 3.167 
                 3.094 
                 4.223 
                 3.1 
                 0.0 
               
               
                   
                 2 
                 1.667 
                 1.829 
                 1.942 
                 1.7 
                 0.1 
               
               
                   
                 4 
                 1.617 
                 1.667 
                 1.578 
                 1.6 
                 0.0 
               
               
                   
                 8 
                 2.763 
                 1.882 
                 2.062 
                 2.3 
                 0.4 
               
               
                   
                 24 
                 0.755 
                 1.175 
                 0.906 
                 1.0 
                 0.2 
               
               
                 DDAIP 
                 0 
                 0 
                 0 
                 0 
                 0.0 
                 0.0 
               
               
                   
                 0.5 
                 13.017 
                 11.983 
                 10.804 
                 12.5 
                 0.5 
               
               
                   
                 1 
                 12.399 
                 10.938 
                 11.891 
                 11.7 
                 0.7 
               
               
                   
                 2 
                 17.564 
                 22.256 
                 20.427 
                 19.9 
                 2.3 
               
               
                   
                 4 
                 17.55 
                 19.542 
                 20.66 
                 18.5 
                 1.0 
               
               
                   
                 8 
                 23.776 
                 25.873 
                 25.143 
                 24.8 
                 1.0 
               
               
                   
                 24 
                 15.818 
                 14.976 
                 21.433 
                 15.4 
                 0.4 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE VI 
               
             
            
               
                   
               
               
                 Derivation of Pharmacokinetic Parameters from the SC Data 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Formulation 
                 Parameter (units) 
                 R-1 
                 R-2 
                 R-3 
                 Mean 
                 SEM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 10% PEG 400 
                 C max  (ng/mL) 
                 10.9 
                 12.2 
                 12.8 
                 12.0 
                 0.68 
               
               
                   
                 T max  (hr.) 
                 0.5 
                 0.5 
                 0.5 
                 0.5 
                 0.00 
               
               
                   
                 AUC (ng * hr/mL) 
                 48.9 
                 44.4 
                 45.1 
                 46.1 
                 1.70 
               
               
                   
                 Mean Residence 
                 8.1 
                 9.2 
                 8.2 
                 8.5 
                 0.42 
               
               
                   
                 Time (MRT, hr.) 
               
               
                   
                 T½ MRT  (hr.) 
                 5.6 
                 6.4 
                 5.7 
                 5.9 
                 0.29 
               
               
                 40% DDAIP 
                 C max  (ng/mL) 
                 23.8 
                 25.9 
                 25.1 
                 24.9 
                 0.75 
               
               
                 HCl 
                 T max  (hr.) 
                 8 
                 8 
                 8 
                 8.0 
                 0.00 
               
               
                   
                 AUC (ng * hr/mL) 
                 459.1 
                 484.7 
                 529.8 
                 491.2 
                 25.32 
               
               
                   
                 Mean Residence 
                 11.4 
                 10.8 
                 12.2 
                 11.5 
                 0.47 
               
               
                   
                 Time (MRT, hr.) 
               
               
                   
                 T½ MRT  (hr.) 
                 7.9 
                 7.5 
                 8.4 
                 7.9 
                 0.33 
               
            
           
           
               
               
               
               
               
               
            
               
                 Bioequivalence 
                 9.4 
                 10.9 
                 11.8 
                 10.7 
                 0.85 
               
               
                 (AUC 40% DDAIP•HCL /AUC 20% PEG ) 
               
               
                   
               
               
                 T½ MRT  = 0.693 * MRT 
               
            
           
         
       
     
     In another embodiment, paclitaxel (Taxol®) was formulated in either 10% polyethylene glycol 300 (PEG 300) in water, or in aqueous solution of DDAIP-HCL (40%). Rats were dosed (5 mg/kg) by oral gavage (PO), as above, with the test composition in the different formulations as a single bolus and blood plasma levels were determined. A 30-fold increase in AUC was achieved with the aqueous solution of DDAIP-HCL over that achieved with the PEG 300 formulation, with no inhibition of P-glycoprotein (P-gp) up to 10 mM. The results are graphically represented in  FIG. 3 . 
     Example II 
     Two small molecule therapeutic compounds were selected from each of BCS Class 2, BCS Class 3, and BCS Class 4 for oral delivery (PO) to jugular vein cannulated (JVC) male Sprague Dawley rats (200-250 gram weight). The BCS Class 2 compounds were haloperidol and sulfasalazine. Haloperidol is an antipsychotic butyrophenone sold under the brand name Haldol® and sufasalazine is an anti-inflammatory sulfa drug derivative of mesalazine sold under the brand name Azulfidine®. The BCS Class 3 compounds were atenolol, and glucosamine (in the salt form glucosamine sulfate). Atenolol is in the class of beta blocker drugs and is sold under the brand names Senormin® and Tenomin. The BCS Class 4 compounds were furosemide (Lasix®) and chlorothiazide (Diuril®), both of which are diuretics. 
     The compounds from BCS Class 2, 3 and 4 were each formulated at 5 mg/ml in a liquid carrier of either an aqueous solution of polyethylene glycol 400 (20% PEG 400), or an aqueous solution of DDAIP-HCL (20%). The BCS Class 2 compounds, haloperidol and sulfasalazine, formed suspensions in either liquid vehicle. The BCS Class 3 compounds, atenolol and glucosamine sulfate formed suspensions in the PEG 400 vehicle and solutions in the water vehicle. The BCS Class 4 compounds, furosemide and chlorothiazide formed suspensions in the PEG400 vehicle and solutions in the water vehicle. 
     Cohorts of three animals per group were placed in the study. On day 1, animals from each group were dosed at 30 mg/kg (6 ml/kg) by oral gavage (PO). Animals were fed at the time of dosing. Serial blood samples were collected at pre-dose, and after post-dose intervals of 30 minutes, 1 hour, 2 hours and 4 hours. The blood samples were collected into lithium heparin-coated tubes. At each time point, 0.15 ml of blood was collected via left jugular vein cannulae and processed for collection of plasma by centrifuging at approximately 2,000 rpm for approximately 10 minutes. The cellular fraction of the blood was discarded. Collection times and volume of plasma samples were recorded and tabulated. Plasma samples were transferred into clean tubes and snap frozen onto dry ice. Samples were stored at −80° C. until ready for bio-analysis by LC/MS/MS. The results are tabulated in Table VII. The drug concentration was measured from supernatant after centrifugation of the dosing solution 
     
       
         
           
               
             
               
                 TABLE VII 
               
             
            
               
                   
               
               
                 Oral Formulations - Solubility and AUC 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Aqueous 
                   
                   
                   
                   
                   
               
               
                   
                   
                 Dosing 
                 Drug 
                   
                   
                 Sol.*** 
                 Sol. 
               
               
                   
                 Compound 
                 solution 
                 Conc. 
                 Sol.* 
                   
                 H 2 O 
                 Over 
               
               
                 BCSCl 
                 5 mg/ml 
                 (20%) 
                 mg/ml 
                 over PEG 
                 AUC 0-24 h** 
                 mg/ml 
                 H 2 O 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 2 
                 Haloperidol 
                 PEG 400 
                 1.43 
                 — 
                 1.3 
                 0.001 
                 1430 
               
               
                   
                   
                 DDAIP•HCL 
                 1.73 
                   
                   
                   
                 1730 
               
               
                 2 
                 Sulfasalazine 
                 PEG400 
                 &lt;0.01 
                 &gt;446 
                 24 
                 0.010 
                 0 
               
               
                   
                   
                 DDAIP•HCL 
                 4.46 
                   
                   
                   
                 446 
               
               
                 3 
                 Glucosamine 
                 PEG 400 
                 4.52 
                 — 
                 2.5 
                 551.000 
                 — 
               
               
                   
                   
                 DDAIP•HCL 
                 4.42 
                   
                   
                   
                 — 
               
               
                 3 
                 Atenolol 
                 PEG 400 
                 2.74 
                 2.300 
                 2.1 
                 13.500 
                 — 
               
               
                   
                   
                 DDAIP•HCL 
                 6.28 
                   
                   
                   
                 — 
               
               
                 4 
                 Furosemide 
                 PEG 400 
                 &lt;0.01 
                 &gt;400 
                 9.1 
                 0.006 
                 0 
               
               
                   
                   
                 DDAIP•HCL 
                 4.00 
                   
                   
                   
                 667 
               
               
                 4 
                 Chlorothiazide 
                 PEG 400 
                 &lt;0.01 
                 &gt;363 
                 18 
                 0.300 
                 0 
               
               
                   
                   
                 DDAIP•HCL 
                 3.63 
                   
                   
                   
                 12 
               
               
                   
               
               
                 Notes to Table VII. 
               
               
                 *Solubility improvement fold over PEG 400 
               
               
                 ** UC-improvement fold for hourly period 
               
               
                 ***Reported solubility from the literature 
               
               
                 ****Solubility improvement fold over water. 
               
               
                 *****Pharmacokinetic parameters were calculated with PK Solutions 2.0 software (Summit Research Services, Montrose, CO) 
               
            
           
         
       
     
     The results show that the DDAIP-HCL improves the solubility of the compounds when compared to water or PEG vehicle. This improvement is also reflected in the enhanced blood levels of drug in the rat plasma. In particular, the Class 2 and 4 compounds, where solubility is a determining factor, displayed over a 400-fold solubility improvement and over a 20-fold AUC improvement. The data show that formulations with DDAIP.HCL significantly improve the oral delivery of difficult-to-absorb small molecule therapeutic compounds by improving their solubility and, hence, their absorption. 
     Example III 
     Male CD1 mice were used (Harlan, USA) in this study and weighed 20-24 grams at the time of use. A 1.5% aqueous methylcellulose solution was made overnight with continuous heating and stirring. To this, 50 mg Phenol Red was added to 100 ml of 1.5% aqueous methylcellulose. Mice were pre-treated with saline or 20% DDAIP free base (5 ml/kg) with and without lansoprazole (10 mg/kg) 15 minutes before challenge with Phenol Red at T 0  (150 μl/mouse) Ten and thirty minutes after Phenol Red challenge, mice were euthanized with isoflurane and the stomachs rapidly excised (clamping the pyloric and cardiac sphincters to avoid loss of contents). Stomachs were then cut into several pieces and placed in 15 ml tubes containing 2 ml water prior to processing for A 558  nm measurements. Several mice were sacrificed immediately after gavage with dye to act as an indicator of maximum dye retrieval (75 μg dye was dosed to each mouse). 
     Data (n=3) for stomach emptying (i.e., the amount of dye remaining in the stomach) 10 and 30 minutes after challenge are shown in  FIG. 4 . Pre-treatment with DDAIP slowed stomach emptying compared to pre-treatment with saline at both time points. The same pattern was observed when lansoprazole was administered in either vehicle. Mice that were challenged with dye and immediately sacrificed had stomachs that contained virtually all the administered amount of dye (75 μg). This experiment shows that increased absorption is not due to faster gastric emptying so that, in addition to improvement in solubility, protection of API in stomach and/or increased absorption in the stomach and intestine are other likely mechanisms. 
     Example IV 
     A total of nine male Beagle dogs (three groups of n=3 with body weights ranging from 9.4-10.6 kg) were used from Bio-Quant&#39;s colony (originally sourced from Marshall Farms, North Rose, N.Y., USA) and fasted overnight prior to an intramuscular injection of pentagastrin (6 μg/kg at 0.03 ml/kg) one hour before treatment with lansoprazole (15 mg per animal). Pre-treatment blood samples were taken prior to intramuscular injection with pentagastrin (−60 minutes) and treatment with lansoprazole (time 0). Dogs were then treated with lansoprazole as detailed in Table VIII below. 
     
       
         
           
               
             
               
                 TABLE VIII 
               
               
                   
               
               
                 Experimental Grouping Table 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Group 
                   
                   
                   
                   
                   
                 Overnight 
               
               
                 (n = 3) 
                 Pre-Treatment 
                 Dose 
                 Route 
                 Regimen 
                 Bleeds 
                 Fasting 
               
               
                   
               
               
                 1, 2, 3 
                 Pentagastrin 
                 60 μg/kg at 
                 I.M. 
                 Single 
                 Predose 
                 Yes 
               
               
                   
                   
                 0.03 ml/kg 
                   
                 (−60′) 
                 (−60′) 
               
               
                   
               
               
                   
                   
                 Dose per 
                   
                   
                   
                 Overnight 
               
               
                   
                 Treatment 
                 Dog 
                 Route 
                 Regimen 
                 Bleeds 
                 Fasting 
               
               
                   
               
               
                 1 
                 Lansoprazole 
                 15 mg 
                 Oral gavage 
                 1 capsule 
                 Pre-dose, 
                 Yes 
               
               
                   
                 powder in 1 
                   
                 with 
                 per dog 
                 30 min, 1 hr, 
               
               
                   
                 gelatin capsule 
                   
                 Petfiller 
                   
                 2 hr, 4 hr, 
               
               
                 2 
                 Lansoprazole 
                 15 mg 
                 followed by 
                 1 capsule 
                 8 hr, 24 hr. 
               
               
                   
                 powder in 1 
                   
                 50 ml H 2 O 
                 per dog 
                 post- 
               
               
                   
                 gelatin capsule 
                   
                   
                   
                 dosing 
               
               
                   
                 containing 
               
               
                   
                 0.625 g of 100% 
               
               
                   
                 DDAIP•base 
               
               
                 3 
                 Lansoprazole 
                 15 mg 
                   
                 5 capsules 
               
               
                   
                 powder in 5 
                   
                   
                 per dog 
               
               
                   
                 gelatin capsules, 
               
               
                   
                 each containing 
               
               
                   
                 0.625 g of 100% 
               
               
                   
                 DDAIP•base 
               
               
                   
               
            
           
         
       
     
     Formulations: 
     
         
         Group 1: Dosed with 15 mg lansoprazole powder in a single gelatin capsule (size 00). 
         Group 2: 700 μl (0.625 g) DDAIP*base was mixed with 15 mg of lansoprazole powder in a single gelatin capsule (size 00). 
         Group 3: 700 μl (0.625 g) DDAIP*base was mixed with 3 mg of lansoprazole powder in each of 5 gelatin capsules (size 00). 
       
    
     Blood Collection: 
     At each time point shown in Table VIII above, 0.5 ml whole blood samples were collected from the saphenous vein into heparinized blood collection tubes. Following centrifugation at 10,000 rpm at 4° C. for 10 minutes, plasma samples were collected and stored at −80° C. until analysis by LCMS-MS. 
     RESULTS AND CONCLUSION 
       FIG. 5  shows the pharmacokinetic profiles after oral treatment with lansoprazole in all groups. Table IX details individual parameters, such as T max , C x  and TH. In comparison to treatment with lansoprazole powder alone, the addition of DDAIP (0.625 g in 12 capsule or 3.125 g in a total of capsules) resulted in higher C max , longer T½ (Table IX) and significantly increased AUC values. T max  occurred at 1 hour in all groups. Furthermore, AUC values were also significantly different in both DDAIP-treated groups. 
     This study illustrates that the pharmacokinetic profile of orally-administered lansoprazole is significantly improved in dogs by the addition of DDAIP to the formulation. In addition, the group treated with 3.125 g DDAIP in five gelatin capsules exhibited significantly improved pharmacokinetic properties when compared to groups treated with control formulation or 0.625 g DDAIP in one gelatin capsule. These improvements are representative of human subjects&#39; dosing since this experiment was performed at physiological pH of the human stomach. 
     
       
         
           
               
             
               
                 TABLE IX 
               
             
            
               
                   
               
               
                 Pharmacokinetic Properties in all Groups 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 C max   
                 T max   
                 T½ 
               
               
                   
                 Group 
                 (ng/ml) 
                 (hr.) 
                 (hr.) 
               
               
                   
               
               
                   
                 1 
                 260 
                 1 
                 0.61 
               
               
                   
                 2 
                 369 
                   
                 1.78 
               
               
                   
                 3 
                 445 
                   
                 1.78 
               
               
                   
               
            
           
         
       
     
     The foregoing discussion and the examples are illustrative, but are not to be taken as limiting. Still other variants within the spirit and scope of this invention are possible and will readily present themselves to those skilled in the art.