COMPOSITION COMPRISING ANTIMICROBIAL AGENT AND ITS USES

Abstract

A composition comprising 2,3 dihydroxypropyl dodecanoate, and an emulsifier results in greater dissolution and biodistribution of 2,3 dihydroxypropyl dodecanoate to enhance antimicrobial and other activities.

Claims

1. A solid form composition comprising: (i) 2,3 dihydroxypropyl dodecanoate, and (ii) an emulsifier.

2. The composition of claim 1 wherein the composition comprises of from about 5 wt. % to about 95 wt. % 2,3 dihydroxypropyl dodecanoate based on the total weight of the composition.

3. (canceled)

4. The composition of claim 1 wherein the composition comprises of from about 2 wt. % to about 95 wt. % emulsifier based on the total weight of the composition.

5. (canceled)

6. The composition of claim 1 wherein the emulsifier selected from the group consisting of Fatty acids, Glyceryl-lacto esters of fatty acids, salts of fatty acids, mono glycerides of fatty acids, Lactylated fatty acid esters of glycerol and propylene glycol, Lactylic esters of fatty acids, sucrose oligoesters, sorbitol, polysorbitan, Lecithin, hydroxylated Lecithin, and Sodium Lauryl Sulfate, or combinations thereof.

7. (canceled)

8. The composition of claim 1 wherein the composition comprises one or more emulsifiers selected from the group consisting of short chain monoglycerides, short chain fatty acids, glyceryl polyethyleneglycol ricinoleate, and sucroglycerides or combinations thereof.

9. The composition of claim 1 wherein the composition comprises one or more emulsifiers selected from the group consisting of those obtained from plant extracts, for example those extracted from Quillaia, Yucca or seaweed.

10. The composition of claim 1 wherein the 2,3 dihydroxypropyl dodecanoate is encapsulated in the emulsifier.

11. The composition of claim 1 wherein the emulsifier is encapsulated in the 2,3 dihydroxypropyl dodecanoate.

12. (canceled)

13. (canceled)

14. (canceled)

15. The composition of claim 1 wherein the composition is in the form of a solid nanoemulsion.

16. (canceled)

17. (canceled)

18. The composition of claim 1 wherein the composition is coated with a polysaccharide, for example maltodextrin or inulin.

19. (canceled)

20. The composition of claim 1 wherein a single oral dose of the composition at a dosage of 0.025 g/kg body weight causes blood plasma levels of 2,3 dihydroxypropyl dodecanoate to exceed 3.9 micrograms/ml peak concentration within 6 hours.

21. The composition of claim 1 wherein dosing of the composition at a dosage of 0.025 g/kg body weight every 12 hours causes extracellular fluid in the lungs to have a concentration of 2,3 dihydroxypropyl dodecanoate which exceeds 3.9 micrograms/ml peak after 92 hours.

22. The composition of claim 1 wherein the composition further comprises a further pharmaceutically active agent.

23. The composition of claim 1 wherein the composition is microstatic and/or the composition is microcidal.

24. (canceled)

25. The composition of claim 1 wherein the composition further comprises one or more nutraceuticals.

26. (canceled)

27. (canceled)

28. (canceled)

29. (canceled)

30. A nutraceutical composition comprising the composition of claim 1.

31. A food composition comprising the composition of claim 1.

32. (canceled)

33. The composition of claim 1 for use as an anti-microbial agent.

34. The composition of claim 1 for use in the treatment of respiratory tract infections in mammals.

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)

40. (canceled)

41. The compound 2,3 dihydroxypropyl dodecanoate for use in the treatment of infectious disease such as respiratory tract infections.

42. A method of treatment of infectious disease such as respiratory tract infections in an animal subject such as a human subject in need thereof, comprising administering to the subject the compound 2,3 dihydroxypropyl dodecanoate.

43. A method of treatment of infectious disease such as respiratory tract infections in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

44. A method of treatment of gastrointestinal upset, for example chronic diarrhoea in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

45. A method of modulating gastrointestinal microbiota in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

46. A method of modulating triglyceride metabolism, for example high density lipoprotein and/or low-density lipoprotein metabolism, in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

47. A method of reducing bilirubin levels in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

48. A method of increasing alkaline phosphate levels in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

49. A method of decreasing urea levels in an animal subject such as a human subject in need thereof, comprising administering to the subject the composition of claim 1.

50. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

[0088] The following examples are not intended to limit the present application.

Examples

[0089] 1800 pigs weighing from 40 to 50 kg body weight (BW) were diagnosed with both porcine reproductive and respiratory syndrome (PRRS) and influenza positive (Human variant swine flu H1N1) concurrently. The pigs were showing clinical signs of fever, coughing, poor appetite and high mortality.

[0090] The pigs were randomly assigned to three groups:

[0091] 1) Positive control group (600 pigs) received 0.08 g/kg BW of a commercial formulation of 2,3 dihydroxypropyl dodecanoate on a silica carrier in feed;

[0092] 2) LDF1 low dose group (600 pigs) received 0.015 g/kg BW of a composition of the invention in liquid form comprising 6.5 wt. % 2,3 dihydroxypropyl dodecanoate (based on the total weight of the composition) and an emulsifier in feed;

[0093] 3) LDF1 High dose group (600 pigs) received 0.025 g/kg BW of a composition comprising of the invention in liquid form comprising 6.5 wt. % 2,3 dihydroxypropyl dodecanoate (based on the total weight of the composition) and an emulsifier in feed.

[0094] The pigs were monitored daily over 14-day period to assess symptoms on a group basis. A Daily score of symptom severity within each group was rated from 0 to 5 for each group on each day based on 15-minute observation of coughing. Coughing is a symptom of PRRS and influenza. The more pigs who cough means the more animals who currently exhibit symptoms of these diseases and provides a good representation of the overall health of each group.

[0095] Score 0 represents numerically no animals coughing that is, no animals from the 600 pigs in each group coughed during the 15 minutes observation. A score of 1 means a single pig in 600 coughed during the 15-minute observation period, a score of 2 score means two pigs coughed during the 15-minute observation period, a score of 3 means three pigs coughed during the 15-minute observation period, a score of 4 means 4 pigs coughed during the 15-minute observation period, a score of 5 means five or more animals coughed during the 15-minute observation period. Daily mortality for each group was recorded. The results are shown in Table 4. As can be seen the pigs who were treated with the composition of the present invention coughed less often. The pigs who were treated with the composition of the invention showed less symptoms of disease and were in overall better heath than those treated with the control composition comprising 2,3 dihydroxypropyl dodecanoate without an emulsifier.

Results

[0096]

TABLE-US-00005 TABLE 4 Control Control 0.08 g/kg LDF1 LDF1 0.08 g/kg LDF1 LDF1 (unenhanced) 0.015 g/kg 0.025 g/kg (unenhanced) 0.015 g/kg 0.025 g/kg Symptom Symptom Symptom No. Deceased No. Deceased No. Deceased Day score score score pigs/d pigs/d pigs/d 1 4 4 4 0 0 0 2 4 4 4 2 1 0 3 4 2 2 1 0 1 4 3 2 1 3 0 0 5 3 1 0 1 0 0 6 4 1 1 2 1 1 7 4 1 1 0 1 1 8 4 0 0 1 0 0 9 3 1 1 2 3 1 10 4 2 0 3 2 1 11 4 3 0 1 2 2 12 4 1 1 1 1 1 13 4 1 1 2 0 0 14 4 1 0 2 0 0 AVE 3.785714 1.714286 1.142857 1.5 0.785714 0.571429 N = 1800 pigs, selected at random to positive control group (n = 600) or LDF1 low dose (n = 600) or LDF1 high dose (n = 600); “pigs/d” is pigs per day.

[0097] At the start and end of the trial 5 pigs from the control group and 5 pigs from the LDF1 High dose group were tested for PRRS antibodies by ELISA and presence of PRRS RNA by PCR. In brief, blood was drawn from 5 pigs selected at random from each group on day 1 of the trial. The blood was tested for the presence of PRRS antibodies by ELISA and presence of PRRS RNA by PCR. Blood was drawn from 5 pigs selected at random from each group on day 14 of the trial. The blood was tested for the presence of PRRS antibodies by ELISA and presence of PRRS RNA by PCR. On day 1 all animals tested positive for both PRRS antibodies and PRRS RNA. On day 14 no animal in the treatment group tested positive for PRRS RNA indicating that there was no viral load in these animals that is, the PRRS virus was no longer present in the animals. 4 from 5 animals from the control group tested positive for PRRS RNA indicating that these animals were still infected with the PRRS virus. The results of the testing for PRRS antibodies by ELISA and presence of PRRS RNA by PCR are shown in Table 5.

TABLE-US-00006 TABLE 5 Control LDF1 High dose LDF1 High ELISA+/− ELISA+/− Control PCR dose PCR Day 0 5/5 positive 5/5 positive 5/5 positive 5/5 positive Day 14 5/5 positive 5/5 positive 4/5 positive 0/5 positive N = 20 pigs (5 animals selected at random from each group at d 0 and d 14 of trial)

Open-Label, Randomized, Single-Dose, Five-Period, Crossover, Oral Bioavailability Study in Humans to Evaluate the Biodistribution of 2,3 Dihydroxypropyl Dodecanoate and Vitamin D.

[0098] Subjects: 30 health male Healthy male volunteers, as evaluated by medical history, vitals and general clinical examination, of 18 to 45 years (both years inclusive) with BMI of 18.50-29.99 Kg/m.sup.2 were enrolled. Subjects were evaluated for normal or clinically insignificant biochemical, hematological, urinary, serology, Chest X Ray and ECG values/reports. Subjects were generally healthy as documented by the medical history, physical examination (including but may not be limited to an evaluation of the cardiovascular, gastrointestinal, respiratory, musculoskeletal and central nervous systems) and vital sign assessments. Subjects had no evidence of medical illness during screening and check-in. Screening was performed within 28 days of check in. Subjects had negative urine test for drugs, and negative alcohol breathe analysis. Subjects exhibited no evidence of suspected Covid-19 symptoms. Subjects were excluded based on the following criteria: [0099] History of any major surgical procedure in the past 3 months. [0100] History of any clinically significant cardiac, gastrointestinal, respiratory, hepatic, renal, endocrine, neurological, metabolic, psychiatric, hematological and/or any major surgical procedure in the past three months. [0101] History of chronic alcoholism/chronic smoking/drug of abuse/Hypersensitivity. [0102] Subject who consumed tobacco containing products within 48 hours prior to proposed time of dosing [0103] Present or past history of intake of drugs or any prescription drug or over the counter (OTC) drugs within 7 days which potentially modify kinetics/dynamics of 2,3 dihydroxypropyl dodecanoate or any other medication judged to be clinically significant by the investigator. [0104] Consumption of grapefruit and/or its products within 10 days prior to the start of study. [0105] Subject who had participated in any other clinical study or who had bled during the last 90 days. [0106] Subjects who are allergic to coconut/coconut containing foods or known hypersensitivity to 2,3 dihydroxypropyl dodecanoate/lauric acid or its derivatives [0107] History of difficulty in swallowing. [0108] High blood pressure and asthma [0109] Renal or liver impairment [0110] Subjects, who consumed raw coconut, coconut oil, coconut containing products, etc. in last 5 days were excluded from the study because coconut is a natural source of 2,3 dihydroxypropyl dodecanoate. [0111] History of past and present COVID-19 infection

[0112] Study Design

[0113] Subjects were randomly assigned to 5 cohorts with six subjects per cohort. The dosing schedule for each cohort is shown in Table 6. A 7-day washout period was provided between dosing. The formulations are described in Table 7. The dose strength of 2,3 dihydroxypropyl dodecanoate was 1.1 grams delivered orally in either powder form (R, C), liquid form (T1) granule form (T2), or granules in an enteric capsule (T3). The components of each formulation are listed in Table 8a and 8b. Reference formulation (R) is 2,3 dihydroxypropyl dodecanoate which is not in combination with an emulsifier. Comparator (C) formulation, Lauricidin®, is an example of a commercially available 2,3 dihydroxypropyl dodecanoate (monolaurin) powder formulation and is available from Med-Chem Labs, AZ, USA. T1 is a liquid dosage formulation comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier. T2 is 2,3 dihydroxypropyl dodecanoate and an emulsifier in a solid form which is a granule form. T3 is 2,3 dihydroxypropyl dodecanoate and an emulsifier in a solid form which is a granule form encapsulated in an enteric capsule. Doses of each formulation were administered according to the dosing method in Table 9.

[0114] Dosing was performed after overnight fasting of at least 10.00 hours, in the morning a single oral dose of test product (T1, T2, T3, R or C) were administered (according to the randomization schedule and a minimal coconut free food was given prior to dosing) with final volume of 240±02 mL of water at ambient temperature, to the subjects, in seated upright posture, under the supervision of Investigator/medical officer. Compliance with dosing was confirmed by mouth check of the subjects with the help of tongue depressor and torch light to assess compliance to dosing. Subjects remained seated in upright position for at least 04.00 hours of post dose in each period and only necessary movement will be allowed during this period. Subjects were not allowed to lie down (except as directed by the physician secondary to adverse events) during this restriction period. Thereafter, subjects were allowed to ambulate freely during the remaining period of the study. The subjects did not take part in any strenuous exercise/activity during the study. Drinking water was restricted at least 01.00 hour prior to dosing until 01.00 hour post-dose (except 240±02 mL of water given during dosing). At all other times, drinking water was provided freely. Standard meals were provided at 04.00, 09.00 and 13.00 hours post-dose.

TABLE-US-00007 TABLE 6 Study Design Informed Consent & 7-day 7-day 7-day 7-day Screening Washout Washout Washout Washout End of Treatment Period Period Period Period study (6 subjects Week Week Week Week Week per cohort) 1 2 3 4 5 Cohort 1 T1 T2 T3 R C Cohort 2 T2 T3 R C T1 Cohort 3 T3 R C T1 T2 Cohort 4 R C T1 T2 T3 Cohort 5 C T1 T2 T3 R

TABLE-US-00008 TABLE 7 Colour/ Storage Formulation Treatment Dose Strength Form Shape condition Reference (R) 2,3 dihydroxypropyl 1.1 Gram ± 1% Granule White 15° to 25° C. dodecanoate Granule granules Comparator (C) 2,3 dihydroxypropyl 1.1 Gram ± 1% Granule White 15° to 25° C. dodecanoate Granule Lauricidin ® Test (T1) Liquid 15 ml equivalent to Solution Purple 15° to 25° C. formulation about 1.1 grams of coloured containing 2,3 2,3 dihydroxypropyl solution dihydroxypropyl dodecanoate dodecanoate Solution (7.5%) & emulsifier Test (T2) Solid dosage 2.4 Grams ± 1% of Granule White 15° to 25° C. formulation granules equivalent granule containing to about 1.1 grams granules of 2,3 of 2,3 dihydroxypropyl dihydroxypropyl dodecanoate dodecanoate & emulsifier Test (T3) Enteric dosage 4 Enteric Capsules Granule- 00 size 15° to 25° C. formulation containing 600 mg filled White containing granules, Capsule capsule granules of 2,3 equivalent to a dihydroxypropyl total of about 1.1 dodecanoate & grams of 2,3 emulsifier dihydroxypropyl dodecanoate

TABLE-US-00009 TABLE 8a Formulation Treatment Components Analytical Properties Reference (R) 2,3 dihydroxypropyl 2,3 dihydroxypropyl >90% 2,3 dodecanoate granule dodecanoate dihydroxypropyl dodecanoate Comparator (C) 2,3 dihydroxypropyl 2,3 dihydroxypropyl >90% 2,3 dodecanoate (Lauricidin ®) dodecanoate dihydroxypropyl dodecanoate Test (T1) Liquid formulation containing Water, Mono and Di- 2,3 dihydroxypropyl 2,3 dihydroxypropyl Glycerides (2,3 dodecanoate: 1.1 g dodecanoate Solution (7.5%) & dihydroxypropyl Zinc: 10 mg emulsifier dodecanoate), Emulsifiers Vitamin C: 80 mg (Sunflower Lecithin, Vitamin E: 12 mg Polysorbate 80, Vitamin D3: 25 μg Hydrogenated Castor Oil), Vitamin B12: 2.5 μg Sweeteners (Sorbitol, Sucralose), Mixed Berry Extract, Acidity Regulator (Citric Acid, Lactic Acid), Flavours, Natural Flavours, Vitamin C (L- ascorbic acid), Vitamin E (Tocopherols), Zinc Gluconate, Vitamin D3 (Cholecalciferol), Vitamin B12 (Cyanocobalamin) Test (T2) Solid dosage formulation Mono and Di-Glycerides 2,3 dihydroxypropyl containing granules of 2,3 (2,3 dihydroxypropyl dodecanoate: 1.1 g dihydroxypropyl dodecanoate dodecanoate), Emulsifier Vitamin D3: 2000 IU & emulsifier (Sodium Lauroyl Lactylate), Inulin, Vitamin D3 (Cholecalciferol) Test (T3) Enteric dosage formulation Mono and Di-Glycerides 2,3 dihydroxypropyl containing granules of 2,3 (2,3 dihydroxypropyl dodecanoate: 1.1 g dihydroxypropyl dodecanoate dodecanoate), Emulsifier Vitamin D3: 2000 IU & emulsifier (Sodium Lauroyl Lactylate), Inulin, Vitamin D3 (Cholecalciferol), Hypromellose Capsule

TABLE-US-00010 TABLE 8b R C T1 T2 T3 Component Weight percent of total composition 2,3 dihydroxypropyl >93*     >93*     7.353 45.275 45.482 dodecanoate Sodium Lauroyl Lactylate 0.000 0.000 0.000 22.503 22.606 Cabosil (silica) 0.000 0.000 0.000 20.385 20.478 Inulin 0.000 0.000 0.000 10.920 10.970 vitamin D 2MIU/g 0.000 0.000 0.004 0.098 0.099 Cabosil (silica) 0.000 0.000 0.000 0.364 0.366 Poly(methacylic acid-co-methyl 0.000 0.000 0.000 0.455 0.000 methacrylate) 1:2 Castor oil 0.000 0.000 1.470 0.000 0.000 Polysorbate 80 0.000 0.000 2.940 0.000 0.000 Monoglyceride blend 0.000 0.000 7.353 0.000 0.000 Chain length 3, 4, 8, 10 Vit E 98% DL alphatocopherol 0.000 0.000 0.094 0.000 0.000 Lecitas 4719 0.000 0.000 5.880 0.000 0.000 Water 0.000 0.000 68.120 0.000 0.000 Ascorbic acid 0.000 0.000 0.627 0.000 0.000 Zinc Gluconate 0.000 0.000 0.460 0.000 0.000 Vit B12 0.000 0.000 0.020 0.000 0.000 Sorbitol 70% syrup 0.000 0.000 3.676 0.000 0.000 Berry Extract 0.000 0.000 1.765 0.000 0.000 sucralose 0.000 0.000 0.196 0.000 0.000 Total 100     100     100 100 100 *the remainder of the composition comprises derivatives of 2,3 dihydroxypropyl dodecanoate such as lauric acid.

TABLE-US-00011 TABLE 9 Reference (R) 1.1 ± 1% Grams of powder/granule were weighed in a disposable container and given to subject for dosing. Subject was requested to swallow/wash down completely with 240 ml ± 02 ml of water at room temperature. A small volume of water was used for rinsing the container and the subject drank the rinsed solution. It was ensured that no residue remained in the disposable container or in the mouth cavity. This was followed by assessment of Compliance for Dosing. Comparator (C) 1.1 ± 1% Grams of powder/granule was weighed in a disposable container/spatula and given to subject for dosing. Subject was requested to swallow/wash down completely with 240 ml ± 02 ml of water at room temperature. A small volume of water was used for rinsing the container and the subject drank the rinsed solution. It was ensured that no residue remained in the disposable container or in the mouth cavity. This was followed by assessment of Compliance for Dosing. Test (T1) 15 ml of the liquid dose was measured in a measuring container to which 100 ml of water was added (total volume 115 ml) in dispensing container, (care was taken to make sure no sample remained in the measuring cup for each dose), stirred, and given to subject for dosing. Additional 125 ml ± 02 ml water was added to the same container, stirred and subject consumed completely ensuring no residue remains. This was followed by assessment of Compliance for Dosing. Test (T2) 2.4 ± 1% Grams of granule was weighed in a disposable container/spatula and given to subject for dosing. Subject was requested to swallow/wash down completely with 240 ml ± 02 ml of water at room temperature. A small volume of water was used for rinsing the container and the subject drank the rinsed solution. It was ensured that no residue remained in the disposable container or in the mouth cavity. This was followed by assessment of Compliance for Dosing. Test (T3) 4 Capsules were given to the subject and swallowed with 240 ml ± 02 mL water for dosing. This was followed by assessment of Compliance for Dosing.

Sample Collection

[0115] Fecal samples were collected at week 1 and week 2 for all treatment groups during the in-house study period. Fecal sample collected from −12 hrs to 0.00 hrs is considered as pre-dose sample and fecal sample collected after the dosing to 24 hrs is considered as post dose sample. Subject collected stool each time in a labeled disposable container/pack from check in to until 24 hours post dose and stored in the refrigerator (2 to 8° C.). Fecal samples collected from a subject prior the dosing to be pooled, homogenized and transferred in to labeled container. Similarly, all fecal samples collected post dosing from a subject was separately pooled, homogenized and 5 gm of the sample will be transferred into labeled aliquot-I and aliquot-II containers. Aliquots will be stored in a freezer at −70° C.±15° C. until further processing.

[0116] Urine samples were collected at week 1 and week 2 for all treatment groups during the in-house period. Urine samples collected from −12 hrs to 0.00 hrs is considered as pre-dose sample and urine sample collected after the dosing to 24 hrs is considered as post dose sample. Subject collected urine sample each time in a labeled disposable container from check in to until 24 hours post dose and stored in the refrigerator (2 to 8° C.). Urine samples collected from a subject prior the dosing to be pooled, mixed and transferred 10 ml into aliquot-I and aliquot-II containers. Similarly, all urine samples collected post dosing from a subject is to be separately pooled, mixed and 10 ml of the sample will be transferred into labeled aliquot-I and aliquot-II containers. Aliquots were stored in a freezer at −70° C.±15° C. until further processing.

[0117] Blood samples were collected through an indwelling intravenous cannula placed in a forearm vein. Intravenous indwelling cannula was kept in situ as long as possible by injecting about 0.5 mL of 10 IU/mL of heparin in normal saline solution to maintain the cannula patent for collection of the post-dose samples. In such cases blood samples were collected after discarding the first 0.5 mL of heparinized saline containing blood. The pre-dose sample was collected before dosing and post-dose samples was collected after dosing. The blood samples were collected using syringe and/or adaptor and transferred into pre-labelled K2EDTA vacutainers. Vacutainers were placed upright in a rack kept in wet ice bath until centrifugation and during plasma separation. Blood samples were centrifuged at 4000 RPM for 10 minutes at 02° C. to 08° C. to separate the plasma. Centrifugation was started within 30 minutes of the collection of samples. Plasma samples for vitamin D3 (1 mL Pre-Dose 00.00 Hrs and 1 mL Post Dose 12.00 Hrs) analysis were aliquoted and stored in separate vials.

[0118] A total of 17 blood samples were collected per subject per treatment period. The sampling timing is shown in Table 10.

TABLE-US-00012 TABLE 10 Sample Time Points Sample Time Points Volume S. No (hours) (minutes) of Blood 1 −12.00 −720 04 mL 2 −06.00 −360 04 mL 3 00.00 0 (−20 to 0)  05 mL* 4 00.25 +15 04 mL 5 00.50 +30 04 mL 6 00.75 +45 04 mL 7 01.00 +60 04 mL 8 01.25 +75 04 mL 9 01.50 +90 04 mL 10 01.75 +105 04 mL 11 02.00 +120 04 mL 12 02.50 +150 04 mL 13 03.00 +180 04 mL 14 04.00 +240 04 mL 15 06.00 +360 04 mL 16 12.00 +720  05 mL* 17 24.00 +1440 04 mL *Additional 1 mL of blood was collected at time-points 00:00 pre-dose and 12:00 post-dose for vitamin D3 analysis.

Sample Analysis

[0119] 2,3 dihydroxypropyl dodecanoate in plasma, urine & feces samples and vitamin D3 in plasma was assayed using Liquid Chromatography-Tandem Mass Spectorometry (LC-MS/MS). A solution of internal standard in the concentration of 1 μg/mL for Alpha Monolaurin D5 in Acetone-M: Water (70:30, v/v) was prepared. 50 μL of internal standard solution (Alpha Monolaurin D5 1 μg/mL) to 500 μL of samples. Each sample was analysed on a LC-MS/MS system using a Zorbax XDB C18 (100 mm×4.6 mm, 3.5 μm) column with a mobile phase of Acetone-M: Buffer (90:10 v/v) at a column oven temperature of 40° C. MS/MS was performed with a positive mode of ionization. Chromatograms were acquired using the computer-based Analyst software version 1.6.3, supplied by AB Sciex process data by peak area ratio. The concentration of the unknown is calculated from the following equation using regression analysis of spiked plasma calibration standard with the reciprocate of the drug concentration as a weighting factor (1/X.sup.2)

[0120] Y=mx+b

[0121] Where,

[0122] x=Concentration of Alpha Monolaurin

[0123] m=Slope of the calibration curve

[0124] y=Peak area ratio of Alpha Monolaurin to Alpha Monolaurin D5

[0125] b=y−axis intercept of the calibration curve

Statistical Analysis

[0126] Statistical analysis was performed on the pharmacokinetic parameters using SAS®V 9.4. The analysis was performed on data from subjects who complete the entire study.

[0127] Summary Statistics, ANOVA, Ratio analysis, Power, Intra subject CV and 90% Confidence Interval were calculated.

[0128] Descriptive analysis of plasma concentration (time point wise and formulation wise) and pharmacokinetic parameters—Cmax, AUC0-t, and AUCLast were determined for each test formulations. Calculations include the mean, minimum, maximum, range, standard deviation, Standard error, geometric mean and the coefficient of variation for each PK parameters of 2,3 dihydroxypropyl dodecanoate. Ln-transformed data of Cmax, AUC.sub.0-t, and AUC.sub.0-∞ was utilized, when calculating geometric mean and least square ratio.

[0129] Intra-subject variability was computed for Ln-transformed parameters Cmax and AUC.sub.0-t for 2,3 dihydroxypropyl dodecanoate.

[0130] The confidence limits are expressed as a percentage of the least square mean (LSM) of the reference formulation. Using the confidence limits of the above confidence interval and the LSM of the reference product, 90% confidence interval for the ratio of the test and reference product means was calculated.

[0131] The comparison of interest is Test (T) vs Reference (R). Ratios are in the form: —Test/Reference (T/R). Ratio of means was calculated using the LSM of log-transformed pharmacokinetic parameters (Cmax and AUC.sub.0-t). Ratio of means is expressed as a percentage of the LSM of the reference formulation.

[0132] Ratio of geometric least square means for 2,3 dihydroxypropyl dodecanoate of test (T1, T2, T3) and reference (R) formulations was computed for Ln-transformed pharmacokinetic parameters Cmax, AUC.sub.0-t, and AUC.sub.0-∞.

[0133] The power of ANOVA test to detect a 20% mean difference between test formulations was calculated for 2,3 dihydroxypropyl dodecanoate.

Results

[0134] Formulations T1, T2, and T3 comprised vitamin D. Vitamin D (Vitamin D3/Cholecalciferol) was detectable in blood plasma of the subjects after 12 hours post dose. The levels of vitamin D in blood plasma are shown in Table 11. This confirms that the vitamin D of the formulation was bioavailable.

TABLE-US-00013 TABLE 11 75 μg/3000 IU Vitamin D3 ng/ml/3000 IU T3 (Capsule) - Vitamin D3 (ng/ml) - Fasted/Light Meal 4.77 T3 (Capsule) - Vitamin D3 (ng/ml) - Fed/High Fat Meal 3.37 T2 (Granule) - Vitamin D3 (ng/ml) - Fasted/Light Meal 5.65 T2 (Granule) - Vitamin D3 (ng/ml) - Fed/High Fat Meal 7.89 T1 (Emulsion) - Vitamin D3 (ng/ml) - Fasted/Light Meal 7.34 T1 (Emulsion) - Vitamin D3 (ng/ml) - Fed/High Fat Meal 8.83

[0135] The level of 2,3 dihydroxypropyl dodecanoate was determined in urine samples pre and post dosing. The amount of 2,3 dihydroxypropyl dodecanoate was below the level of detection in all samples.

[0136] The level of 2,3 dihydroxypropyl dodecanoate was determined in faecal samples pre and post dosing. The amount of 2,3 dihydroxypropyl dodecanoate was below the level of detection in all samples.

[0137] The levels of 2,3 dihydroxypropyl dodecanoate was determined in blood plasma at 24 hours post dosing. The mean and maximum levels of 2,3 dihydroxypropyl dodecanoate found in blood plasma are shown in Table 12. Dosing performed with a composition comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier provided elevated blood plasma levels of 2,3 dihydroxypropyl dodecanoate after 24 hours. Increased levels of 2,3 dihydroxypropyl dodecanoate may indicate that the composition comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier has dissolved in the gastrointenstinal tract and has avoided lipases in the gastrointestinal tract which would degrade 2,3 dihydroxypropyl dodecanoate. A composition comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier may form micelles such that it undergoes lymphatic uptake and sustained release from the lymphatic system over time.

TABLE-US-00014 TABLE 12 Period 4 & 5 combined Period 3-5 combined Mean Maximum Mean Maximum ng/ml 2,3 dihydroxypropyl ng/ml 2,3 dihydroxypropyl Treatment dodecanoate dodecanoate C 3.376 37.139 2.185 37.139 R 0 0 0 0 T1 23.427 167.383 15.159 167.383 T2 212.368 2463.105 141.579 2463.105 T3 44.669 503.12 29.779 503.12

[0138] The health-promoting effect attributed to a wide range of vitamins, minerals and other moieties is well established. The various formulations relating to the present invention are compatible with moieties exhibiting a broad span of solubility profiles, including water soluble vitamins such as Vitamin C as well as oil-soluble vitamins such as Vitamin D. Importantly, human pharmacokinetic data presented herein demonstrate that all formulations evaluated, namely emulsion (T1), enteric coated granules (T2) and enteric capsule (HPMC) encapsulated granules (T3), result in robust systemic absorption following oral administration. The emulsion form (T1) was not associated with a food effect, systemic bioavailability following oral administration of the enteric capsule encapsuled granules (T3) was higher in the fasted than in the fed state while systemic bioavailability following oral administration of the enteric coated granule form (T2) was greater in the fed than in the fasted state. In summary, there is almost no change in Vit D3 differential levels in High fat meal (P5) when compared to Light meal (P4) for Emulsion (T1), such that Vit D3 absorption remained unchanged (or marginally improved) in T1 group when subjects were fed a high fat high calorie meal; There is an increase in Vit D3 differential levels in High fat meal (P5) when compared to Light meal (P4) for Granules (T2), such that Vit D3 absorption increased in T2 group when subjects were fed a high fat high calorie meal; and there is a decrease in Vit D3 differential levels in High fat meal (P5) when compared to Light meal (P4) for Capsules (T3), such that “Vit D3 absorption decreased in T3 group when subjects were fed a high fat high calorie meal.

[0139] Correlating the pharmacokinetics observed for Vitamin D3 (Vitamin D), the absorption of other oil soluble molecules, including vitamins, minerals and other moieties, including, but not limited to oil-soluble pharmaceutical agents may be similar. This may include various omega oils (3, 6, 9 or other) as well as antioxidants such as Coenzyme 10 (CoQ10), in any of its three redox states, namely fully oxidized (ubiquinone), semiquinone (ubisemiquinone), and fully reduced (ubiquinol), or members of the vanillanoid family, including, but not limited to capsaicin or resiniferatoxin.

[0140] Blood chemistry in all subjects was evaluated pre- and post-dosing in all subjects. As the trial design was a cross over study all subjects had been subject to each formulation.

[0141] It was found that there was a statistically significant increase on alkaline phosphatase (p=0.003) in subjects after completion of the study. As it has been observed that administration of alkaline phosphatase attenuates the inflammatory response and reduces mortality, this may contribute to beneficial effects in acute or chronic infection and related conditions, including but not limited to sepsis, septic shock, acute respiratory distress syndrome, acute lung injury, acute kidney injury, acute liver injury and COVID-19.

[0142] It was found that there was a statistically significant (p=0.0007) elevation in the levels of HDL detected, with no change in LDL levels (p=0.85), a significant reduction in triglyceride levels (p=0.0127) in subjects upon completion of the study. Importantly, the level of very-low-density lipoprotein (VLDL) cholesterol, produced in the liver and released into the bloodstream to supply body tissues with a type of fat (triglycerides), was significantly lower (p=0.0099). Potential benefits include cardiovascular health, including reduced plaque formation.

[0143] It was found that there is a significant reduction in urea levels (p=0.0483) in subjects upon completion of the study. Potential benefits include reducing risk or symptoms of gout and gouty arthritis.

[0144] It was found that there was a statistically significant reduction in bilirubin (Direct) (p=0.0001) in subjects upon completion of the study. Potential benefits include treating or preventing liver disease, particularly hepatitis, anemia or certain drug overdoses.

Dissolution Experiments

[0145] The following experiments were performed to compare the dissolution profiles of the formulations C, R, T1, T2, and T3 as shown in Table 8. Dissolution experiments were performed according to the method of (711) Dissolution USP 2016.

[0146] Dissolution experiment 1: To compare the dissolution of each formulation in 0.1 N HCL the formulation was added to 0.1 N HCL in 900 ml vessels. The vessels were placed in a basket in a standard dissolution bath at a temperature of 37.5° C. The basket was agitated at 75 RPM for 2 hours. The results are shown in Table 12. Only the liquid dosage form comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier showed dissolution after 2 hours.

TABLE-US-00015 TABLE 13 Sample (n = 6) Percent of total sample dissolved at 2 hours C (n = 6) 0 R (n = 6) 0 T1 (n = 18) 67.86667 T2 (n = 6) 0 T3 (n = 6) 0

[0147] Dissolution experiment 2: To compare the dissolution of each formulation in pH 7.2 phosphate buffer each formulation was added to pH 7.2 phosphate buffer in 900 ml vessels. The vessels were placed in a basket in a standard dissolution bath at a temperature of 37.5° C. The basket was agitated at 75 RPM for 10 hours. Dissolution was measured at 1, 4, and 7. The results are shown in Table 13. Only the liquid dosage form comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier showed dissolution above 10% after 1, 4, and 7 hours. C is in pellet form from 2 mm to 5 mm in diameter did not dissolve. R is in granule form with a particle size of from 500-1000 microns and without an emulsifier did show dissolution of less than 10% after 4 and 7 hours.

TABLE-US-00016 TABLE 14 1 hour 4 hours 7 hours Sample Percent of total sample dissolved C (n = 6) 0 0 0 R (n = 6) 0 6.2 9.5 T1 (n = 12) 72.75 74.55 89.15 T2 (n = 6) 0 0 0 T3 (n = 6) 0 0 0

[0148] Dissolution experiment 3: To compare the dissolution of formulation R and T2 in pH 6.8 phosphate buffer containing 0.2% sodium lauryl sulfate (SLS) each formulation was added to 0.1 N HCL in 900 ml vessels. The vessels were placed in a basket in a standard dissolution bath at a temperature of 37.5° C. The basket was agitated at 100 RPM for 6 hours. Dissolution was measured at 2, 4, and 6 hours. The results are shown in Table 14. After 2 hours over 80% of T2 was dissolved indicating greater solubility due to the presence of emulsifiers in the formulation.

TABLE-US-00017 TABLE 15 2 hours 4 hours 6 hours Sample Percent of total sample dissolved R (n = 6) 18.1 82.4 82.9 T2 (n = 6) 84.5 101.4 102.9

[0149] Dissolution experiment 4: To compare the dissolution of each formulation in pH 7.2 phosphate buffer containing 0.2% sodium lauryl sulfate (SLS) each formulation was added to pH 7.2 phosphate buffer containing 0.2% sodium lauryl sulfate (SLS) in 900 ml vessels. The vessels were placed in a basket in a standard dissolution bath at a temperature of 37.5° C. The basket was agitated at 75 RPM for 7 hours. Dissolution was measured at 1, 4, and 7 hours. The results are shown in Table 15. T2 granules are less soluble than T3 granules in gastric resistant capsule. This indicates an effect of Sustained release coating on the granule. This also indicates minimum effect of gastric resistant capsule in pH 6.8. T1, liquid emulsions are readily soluble, >70% at 1 hour and >95% at 7 hours.

TABLE-US-00018 TABLE 16 1 hour 4 hours 7 hours Sample Percent of total sample dissolved C (n = 6) 0 0 0 R (n = 6) 0 6.2 37.4 T1 (n = 12) 72.5 74.55 107.5 T2 (n = 6) 0 0 6.2 T3 (n = 6) 0 0 37.2

[0150] Overall, the dissolution experiments show that 2,3 dihydroxypropyl dodecanoate is poorly soluble in acid and intestinal pH range in the absence of extra emulsifiers either incorporated to the formulation or dissolution media. Smaller particle size promotes dissolution of unformulated 2,3 dihydroxypropyl dodecanoate in the presence of SLS in the media. Liquid emulsified formulations have a rapid dissolution profile in acid and intestinal pH ranges. Gastric resistant capsules did not yield 2,3 dihydroxypropyl dodecanoate in acid conditions but did yield >35% at 7 hours in pH 7.2 with SLS in media. Granules without sustained release coating did not yield 2,3 dihydroxypropyl dodecanoate in acid and yielded only 6.2% at 7 hours at pH 7.2 with SLS media, demonstrating a slow-release profile, relative to granules in capsule that were coated with a sustained release coating.

[0151] The beneficial effects observed in pig studies for the emulsion form when compared to non-formulated or otherwise sub-optimally formulated 2,3 dihydroxypropyl dodecanoate correlates with the dissolution rate of 2,3 dihydroxypropyl dodecanoate observed for various formulations. The emulsion based 2,3 dihydroxypropyl dodecanoate resulted in rapid and complete or near-complete dissolution in both acid and neutral pH conditions, with or without added surfactant. In this embodiment, it is surmised that the effect observed in pigs is related to the enhanced biodistribution of 2,3 dihydroxypropyl dodecanoate in the gastric and intestinal lumen, such effect being due to the intact 2,3 dihydroxypropyl dodecanoate or metabolites or other related degradants thereof.

Case Studies

[0152] Case study 1: Six subjects who had tested positive for COVID 19 by PCR and had current mild to moderate symptoms were instructed to take the 15 ml of the Liquid emulsion product (T1) 3 times daily with food. Ideally diluted in a glass of water 50 ml to 150 ml. The subjects were not screened for any inclusion criteria, other than not requiring hospitalisation and having 2 out of the 3 symptoms fever, sore throat and cough or shortness of breath. No placebo arm or other dosing control was considered for the study. Data collected was qualitative with some objective aspects which translated to symptom scoring as follows,

[0153] Fever Score [0154] a. 0<37.8° C., [0155] b. 1 Reports mild sweats, chills or fatigue and has temperature of 37.8° C. to 38.5° C. [0156] c. 2 Reports moderate discomfort sweats, chills or fatigue and temperature greater than 38.5° C. to 39.0° C. [0157] d. 3 Reports requirement to be in bed and temperature greater than 39.0° C.

[0158] Sore Throat Score [0159] a. 0 No discomfort [0160] b. 1 Reports mild discomfort, able to speak and eat and drink [0161] c. 2 Reports moderate discomfort requiring OTC medications for relief, pain eating drinking and speaking [0162] d. 3 Reports severe discomfort, OTC medications providing some relief, pain causing reduced ability to take fluid and temporary elimination of solid food

[0163] Cough and Shortness of Breath Score [0164] a. 0 No cough or shortness of breath [0165] b. 1 mild occasional coughing or intermittent shortness of breath [0166] c. 2 Persistent dry cough and/or short of breath for prolonged periods (several hours), not necessarily lying in bed [0167] d. 3 Persistent dry cough and/or very short of breath requiring lying in bed

[0168] RESULTS: All subjects reported a reduction in their reported symptoms within 24 or 48 hours. No subjects reported new symptoms or any increased discomfort. The five subjects who reported fever at the start all reported no fever after 24 hours. The results are shown in Table 17.

TABLE-US-00019 TABLE 17 Fever Fever sore throat Sore throat cough cough SUBJECT start 24 hr start 24 hr start 48 hr M01 1 0 2 0 0 0 M02 0 0 2 0 3 1 M03 3 0 2 1 3 1 F01 3 0 3 1 3 1 F02 3 0 2 1 1 0 F03 2 0 1 0 0 0

[0169] CONCLUSION: These data support the conclusion that a composition comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier is effective in the treatment of infectious disease such as respiratory tract infections. The data supports that a method of treatment of infectious disease such as respiratory tract infections in an animal subject such as a human subject in need thereof, comprising administering to the subject the compound 2,3 dihydroxypropyl dodecanoate and an emulsifier is effective.

[0170] Case study 2: Two subjects were provided with the Liquid emulsion formulation (T1) as a potential general health aid and were advised that it may or may not help to prevent symptoms of colds and flus. Surprisingly, both reported an immediate and lasting impact on gut health. Both reported cessation of chronic Diarrhoea.

[0171] SUBJECT 1. Female in her sixties had been suffering chronic diarrhoea for 4 years after her Gall bladder was surgically removed. Her episodes were several times daily and her lifestyle was severely affected. She reported difficulty in performing activities of daily living such as shopping, using public transport, working and exercise, for example swimming.

[0172] She self-administered a dose of 15 ml of Liquid emulsion (T1) with food once daily. After 72 hours her bowel movements had become normal. For five weeks she reported daily normal movements and stool consistency.

[0173] SUBJECT 2. Female in her thirties had been suffering chronic diarrhoea for “at least one year”. There was no obvious trigger or cause. Episodes were daily and seldom had a normal stool consistency.

[0174] She self-administered a dose of 15 ml of Liquid emulsion (T1) with food once daily. After one week she reported her bowel movements had become normal. For five weeks she reported daily normal movements and stool consistency.

[0175] CONCLUSION: The data supports a method of treatment of gastrointestinal upset, for example chronic diarrhoea in an animal subject such as a human subject in need thereof, comprising administering to the subject a composition comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier is effective.

[0176] The composition of the invention comprising 2,3 dihydroxypropyl dodecanoate and an emulsifier, for example in liquid form such as a liquid emulsion, may be an effective aid to improved digestive health in some individuals who suffer chronic diarrhoea.

[0177] The words “comprises/comprising” and the words “having/including” when used herein with reference to the present invention are used to specify the presence of stated features, integers, steps or components but do not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

[0178] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment, within the scope of the appended claims. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination, within the scope of the appended claims.