WO2019102310A1

WO2019102310A1 - Vip formulation for management of inflammatory conditions

Info

Publication number: WO2019102310A1
Application number: PCT/IB2018/058956
Authority: WO
Inventors: Hayley KAPLAN
Original assignee: Alaval Pty Ltd
Current assignee: Alaval Pty Ltd
Priority date: 2017-11-24
Filing date: 2018-11-14
Publication date: 2019-05-31
Anticipated expiration: 2020-05-24

Abstract

The present invention relates to a composition comprising essentially or consisting of: Vasoactive Intestinal Peptide (VIP), a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP has not been chemically modified or conjugated, as well as to a method for manufacturing such a powder or tablet composition and use of the composition in the management of inflammatory diseases or conditions in a subject in need thereof by sublingual administration of the composition.

Description

VIP FORMULATION FOR MANAGEMENT OF INFLAMMATORY CONDITIONS

FIELD OF THE INVENTION

The present invention relates to a composition comprising essentially or consisting of: Vasoactive Intestinal Peptide (VIP), a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP has not been chemically modified or conjugated. The present invention further relates to a method for manufacturing such a powder or tablet composition comprising or consisting of the step of admixing the components with agitation. In addition, the present invention relates to the composition for use in the management of inflammatory diseases or conditions in a subject in need thereof by sublingual administration of the composition.

BACKGROUND OF THE INVENTION

Inflammation is important in the fight against many infections and conditions, but when it becomes persistent, it has been linked to the development of cancers, heart disease and diabetes. Furthermore, various anti-inflammatory medications, if used long term, are associated with significant side-effects such as stomach ulcers and increased risk of bleeding.

Vasoactive Intestinal Peptide (VIP) is a 28 amino acid member of the secretin-glucagon family of peptide hormones (His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-GIn- Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-lle-Leu-Asn-NH2 (SEQ ID NO: 1). Its effects on the immune system are mediated via the adenyl cyclase/ cyclic AMP second messenger system. It is released by nerves in the brain and other sites, particularly around lymph glands. Importantly, immune cells have been found to have a‘lock-and-key’ system specifically for VIP. When acting as the‘key’, VIP has the effect of‘turning off harmful inflammatory signals. However, use of VIP as a therapeutic agent represents a challenge, in that since it is a peptide, if swallowed as a capsule or tablet, VIP would be degraded rapidly by the digestive system and would not reach the blood-stream in its active form. Small peptides such as VIP are notoriously unstable and are susceptible to degradation in aqueous solution. Once VI P has less than 90% of its labelled potency, it is no longer considered to be suitable for administration to a patient. Various types of sugars, surfactants, amino acids and fatty acids, used singly or in combination, have been used in efforts to stabilise VIP in a liquid formulation against degradation which result in modification of the peptide or production of a peptide derivative or conjugate in order to stabilise the peptide.

At present, clinically applicable VIP is formulated as synthetic peptide reconstituted in 0.9% sodium chloride solution (isotonic solution) at a concentration of 0.033 mg/ml_. Alternatively, it has been formulated together with phentolamine by dissolving VIP in a phosphate buffer (pH 2 - 4.5) containing ethylene diaminetetraacetate (EDTA) as stabiliser (EP0493485; US. Pat. No. 5,236,904; US. Pat. No. 5,447,912; WO9505188).

The effects of VIP have been studied in animal models of chronic diseases and conditions, such as rheumatoid arthritis; inflammatory bowel disease and various allergic skin conditions, but these studies have focused on use of a liquid formulation of VIP via the intravenous route, direct injection into joints or via intranasal spray. These modes of delivery of the peptide have thus far limited its use to hospital and research settings. The vasodilatory effects of this peptide have been utilized in the treatment of pulmonary hypertension in human clinical trials. However, in these studies, an aerosol inhaler or nebulizer was used for delivery of a liquid formulation of VIP. VIP has also been delivered in a liquid formulation as a nasal spray for the treatment of biotoxin-related illnesses such as Lyme disease and‘sick-building-syndrome’. T o date over 1800 patients have been treated with the liquid formulation of VIP, and over 98% showed immediate and long-term relief of symptoms such as chronic fatigue, joint pains and even mood disorders.

However, the lack of a simple and effective delivery system for VIP and the complexity of formulation of a liquid dose form of the peptide that requires the addition of stabilisers and other pharmaceutical excipients have limited its development for commercial use, particularly in developing countries.

SUMMARY OF THE INVENTION

According to a first embodiment of the invention, there is provided a composition consisting of or comprising essentially of: Vasoactive Intestinal Peptide (VIP), a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VI P. The VIP may have the amino acid sequence, His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Thr- Arg-Leu-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-lle-Leu-Asn-NH2 (SEQ ID NO: 1), or an amino acid sequence having an equivalent functional efficacy to SEQ ID NO:1 and having 90% sequence identity to SEQ ID NO: 1 , more preferably an amino acid sequence having 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:1. In particular, the VIP has an amino acid sequence identical to SEQ ID NO.1.

The composition may comprise from 0.0023% to 0.0054% (w/w), preferably 0.0040 (w/w) of VIP.

The composition may comprise from 0.83% to 0.86% (w/w) of sugar substitute (preferably sucralose), preferably 0.85% (w/w).

The composition may comprise from 0.12% to 0.15% tartaric acid, preferably 0.14% (w/w).

In particular, the composition may comprise or consist essentially of an admixture of the following ingredients:

sugar substitute (preferably sucralose) 0.85% (w/w)

tartaric acid 0.14% (w/w)

VIP 0.0060% (w/w)

For example, a single dosage of 175 mg may comprise or consist essentially of an admixture of:

145.25 - 150 mg, preferably, 149.4 mg sugar substitute, in particular sucralose;

21 - 26.25 mg, preferably 25 mg tartaric acid; and

0.4 - 0.8 mg, preferably 0.6 mg VIP.

It is to be appreciated that the formulation may optionally comprise additional non-essential ingredients including flavourants, but that such ingredients are not essential elements of the formulation and are not necessary.

According to a further embodiment of the invention, there is provided a method for manufacturing the composition according to the invention as provided above, consisting of or comprising essentially of: VIP, a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VIP, the method comprising or consisting of the steps of admixing the sugar substitute, preferably sucralose, tartaric acid and VIP with agitation. Alternatively, the method may comprise or consist of the steps of admixing the sugar substitute, preferably sucralose, tartaric acid and VIP with agitation, followed by a further step of compression or compaction into a tablet form.

More particularly, the method may comprise the steps of first mixing the sugar substitute, preferably sucralose, and tartaric acid by agitation, followed by addition of the VIP with further agitation. In an alternative embodiment, the sugar substitute, preferably sucralose, and tartaric acid are mixed with the VIP simultaneously by agitation.

According to a further embodiment of the invention, there is provided a composition according to the invention as provided above, consisting of or comprising essentially of: VIP, a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VIP, the compound being for use in the management of inflammatory diseases or conditions in a subject. In particular, the composition may be administered by sublingual administration to a subject in need thereof. In particular, the powder or tablet composition is administered by placing a dose of the powder or tablet composition under the tongue of the subject.

According to a further embodiment of the invention there is provided a method of management of inflammatory diseases or conditions in a subject in need thereof by sublingual administration of a composition of the invention provided above, consisting of or consisting essentially of: VIP, a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet, wherein the VIP is not a chemically modified or conjugated form of VIP. In particular, the powder or tablet composition is administered by placing a dose of the powder or tablet composition under the tongue of the subject.

According to a further embodiment of the invention, there is provided use of VIP, a sugar substitute, preferably sucralose, and tartaric acid in the manufacture of a composition according to the invention as provided above, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VIP.

The sugar substitute is preferably a non-nutritive sweetener, including a sweetener such as sucralose, powdered xylitol, stevia, powdered eryth ritol and others known to the art as derived from plant extracts and/or plant sugars, or any combination thereof, or an artificial sweetener suitable for a diabetic which is synthetically derived such as saccharine, aspartame, or acesulfame K or any combination thereof. Optionally a combination of plant-derived sweetener and artificial sweetener may be used. However, in a preferred embodiment of the invention, the non-nutritive sweetener is sucralose. In particular, the composition may be used for the management of chronic or acute inflammatory conditions such as aches, pains, stiffness and minor joint swelling.

In particular, the composition may be administered as a dosage of 175 mg. More particularly, the composition may be administered twice daily.

DRAWINGS

Figure 1 : shows average pain and stiffness scores for each subject over the duration of the clinical trial;

Figure 2: shows the average levels of inflammation marker IL-6 (pg/ml) in each subject before and after the clinical trial; and

Figure 3: shows a graphical illustration of active VIP concentration measured in serum in pg/ml over time.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to a composition consisting of: VIP, a sugar substitute, preferably sucralose, and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VIP. The present invention further relates to a method for manufacturing such a powder or tablet composition comprising or consisting of the steps of admixing the components with agitation. In addition, the present invention relates to the composition for use in the management of inflammatory diseases or conditions in a subject in need thereof by sublingual administration of the composition.

The applicant has shown that surprisingly, a powdered or compacted or compressed tablet formulation of VIP together with a sugar substitute, preferably sucralose, and tartaric acid combined by agitation in a simple admixture may be used as a sublingual agent for the optimal delivery of active VIP to the immune system. The powdered formulation may be used as is, or compressed or compacted into a tablet form. The VIP raw material may be sourced from any peptide-manufacturing supplier. Of note was that the VIP formulation required no further chemical procedures to be applied to the peptide, viz. no stabilization, modification or conjugation steps with other moieties were required for production of a stable and effective peptide formulation. It is to be appreciated that there is no requirement to formulate the composition by prior dissolution in an aqueous medium for delivery as an aqueous dosage form as would be the case for nasal or aerosol sprays or for injectable delivery.

A“peptide” is a compound consisting of two or more amino acids linked in a chain, the carboxyl group of each acid being joined to the amino group of the next by a bond of the type -OC-NH-

Within the context of this application, a“chemically modified or conjugated form of VIP” means VIP that functions in interaction with other (non-polypeptide) chemical groups attached by covalent bonding or weak interactions as opposed to a VIP that contains only amino acids and no other chemical groups.

The formulation is produced by combining VIP peptide in lyophilised or freeze-dried form with tartaric acid and a sugar substitute, preferably sucralose, by simple mixing and/or agitation methods. The compressed tablet may be formed from the powered mixture by compression of the powdered mixture, a standard method known to those skilled in the pharmaceutical manufacturing field.

Tartaric acid and its derivatives are well known in the art as an anti-oxidant.

The sugar substitute is preferably a non-nutritive sweetener, including a sweetener such as sucralose, powdered xylitol, stevia, powdered eryth ritol and others known to the art as derived from plant extracts and/or plant sugars, or any combination thereof, or an artificial sweetener suitable for a diabetic which is synthetically derived such as saccharine, aspartame, or acesulfame K or any combination thereof. Optionally a combination of plant-derived sweetener and artificial sweetener may be used. However, in a preferred embodiment of the invention, the sweetener is sucralose. Sucralose is a plant sugar derivative that is suitable for use in diabetics and was preferred for use as, unlike other non-nutritive sweeteners tested, it had a more favourable flavour and texture for use in a sublingual delivery composition.

A simple powdered formulation consisting of VIP, tartaric acid and sucralose was shown by the applicant to be suitable for dissolving under the tongue. A person skilled in the art would appreciate that equally a compressed tablet formulation for dissolving under the tongue would work as effectively.

The applicant has demonstrated that a powdered, compacted or compressed tablet formulation of VIP together with a sugar substitute, preferably sucralose, and tartaric acid combined by agitation in a simple admixture and administered sublingually to the subject is effective in the management of inflammatory conditions. EXAMPLES

The invention will now be described with reference to the following illustrative example which should not be construed as to limit the scope of the invention in any way.

Example 1 : Development of the product

Materials and Methods:

Purified, lyophilised VIP was provided as purchased from a peptide manufacturing facility. Tartaric acid and sucralose in powdered form suitable for sublingual administration were purchased.

Sucralose and tartaric acid were first admixed by simple agitation. Thereafter, lyophilised VIP was added to the sucralose and tartaric acid mixture and mixed by simple agitation until homogeneously mixed. The powdered formulation was then filled into a sachet ready for sublingual delivery of the formulation.

Each single-dose sachet was filled to contain a 175 mg dose of the final mixture, consisting of an admixture of:

149.4 mg sucralose;

25 mg tartaric acid; and

0.6 mg VIP.

Example 2: Phase I Clinical Trial

Clinical Data

The Phase I clinical trial was performed on 10 participants recruited with informed consent; 7 women and 3 men, having an average age of 61 yrs (range 50-75).

The average body mass index (BMI) was recorded as 25.7 (range 23-34).

All were non-smokers.

All had normal blood pressure with an average of 120.3/78.3 and none were taking medication for hypertension.

None were on any anti-inflammatory medications.

The average number of joints reported to be affected by pain or stiffness was 3.7.

Performance of Clinical Trial

Each single-dose sachet contained a 175 mg dose of the final mixture, consisting of:

149.4 mg sucralose; 25 mg tartaric acid; and

0.6 mg VIP.

The single-dose sachet powdered formulation was administered sublingually to each subject twice daily (in the morning and evening).

Prior to commencing the trial, all subjects adhered to a‘wash-out’ period of 7-10 days, in which no anti-inflammatories could be taken. Blood samples were taken before starting the trial and after 10 days on the test product.

Each participant was examined by an orthopaedic surgeon on day 1 and day 11 of the trial.

Each participant scored pain and/or stiffness of joints out of 10 (with 10 being highest level of pain and 0 = no pain) on days 1 , 4, 7 and 10 or the trial.

Results No adverse effects were recorded in blood pressure, pulse, weight, blood counts, liver function tests and urine tests, after 10 days on the product.

Average pain scores are shown in Table 1 below and Figure 1.

Table 1 : Average pain scores over the duration of the clinical trial

Clinical Assessment by Orthopaedic Surgeon Days 1 and 11

Table 2 below shows the clinical assessment provided by the orthopaedic surgeon.

Table 2: Clinical assessment

ROM= range of movement

OA= osteoarthritis; RA= Rheumatoid Arthritis

R= right; L=left; AC= acromio-clavicular joint; IP= interphalangeal; MPJ= middle phalangeal joint; CMP= chondromalacia patella; IBS= irritable bowel syndrome. No side-effects or adverse events were documented.

No adverse effects on weight, BMI, blood pressure (systolic and diastolic) or pulse rate were documented.

No adverse effects on subject blood counts were documented. The average results before starting and after taking the 10 day course of test product are shown in Table 3 below.

Table 3: Average blood count

Hb - Haemoglobin

Hot - Haematocrit

RCC - red cell count

WCC - white cell count

Pit - platelet

No adverse effects were recorded for liver function. The average results shown before starting and after taking 10 day course of test product are provided in Table 4 below. Table 4: Average liver function

AST - aspartate aminotransferase

ALT - alanine aminotransferase

ALP - alkaline phosphatase Detection of Inflammation Marker lnterleukin-6 (IL-6)

IL-6 is produced at sites of inflammation and is a key mediator of responses such as stimulation of the liver to produce other inflammatory proteins and activation of white blood cells to produce antibodies.

Levels of IL-6 were determined in the subjects before starting the clinical trial and after completion of 10 days of treatment with the test product. The average results for IL-6 levels (pg/ml) in each subject before and after the clinical trial are shown in the Figure 2.

Summary

Detection of the IL-6 marker for inflammation revealed, on average, a 40% decrease in the level of IL-6.

All subjects receiving test product reported having experienced improvement in their levels of pain and/or stiffness.

Example 3: Pre-clinical test - Evidence for a new delivery mode of action for VIP

Two healthy volunteers were recruited, both fasted for 4 hours and then had baseline blood levels taken.

Volunteer A received a 300 g mixture of powdered formulation of 149.8 mg sucralose, 149.8 mg tartaric acid and 0.4 mg VIP.

Volunteer B received only the sweetener and tartaric acid (1 : 1 , v/v).

Each volunteer dissolved the powdered formulation under the tongue, and blood samples were then taken at 5 time points thereafter, i.e. 1 minute, 2 minutes, 5 minutes, 20 minutes and 30 minutes.

There were no adverse side-effects identified in either patient.

As illustrated in Figure 3, VIP at a concentration of nearly 3000 pg/ml was detected in patient serum at 20 minutes compared with the baseline level of the assay of 1000 pg/ml.

The control patient was negative for VIP at each time point monitored relative to the baseline level.

Claims

1. A composition comprising or consisting of: Vasoactive Intestinal Peptide (VIP), a sugar substitute and tartaric acid, formulated as a powder or tablet for sublingual administration, wherein the VIP is not a chemically modified or conjugated form of VIP.

2. The composition of claim 1 , wherein the sugar substitute is a powdered non-nutritive sweetener, selected from any one or more of sucralose, xylitol, stevia, erythritol, saccharine, aspartame or acesulfame K.

3. The composition of claim 1 , wherein the sugar substitute is a powdered plant-derived non-nutritive sweetener selected from any one or more of sucralose, xylitol, stevia, erythritol.

4. The composition of claim 1 , wherein the sugar substitute is powdered sucralose.

5. The composition of any one of claims 1 to 4, wherein the VIP has the amino acid sequence HSDAVFTDNYTRLRKQMAVKKYLNSILN-NH2 (SEQ ID NO: 1), or an amino acid sequence having an equivalent functional efficacy to SEQ ID NO:1 and having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:1.

6. The composition of claim 5, wherein the VIP has the amino acid sequence His-Ser- Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-GIn-Met-Ala-Val-Lys-Lys-Tyr- Leu-Asn-Ser-lle-Leu-Asn-NH2 (SEQ ID NO: 1).

7. The composition of any one of claims 1 to 6, comprising from 0.0023% to 0.0054% (w/w) of lyophilised VIP.

8. The composition of claim 7, comprising 0.0040 (w/w) of lyophilised VIP.

9. The composition of any one of claims 1 to 8, comprising from 0.83% to 0.86% (w/w) of powdered sugar substitute.

10. The composition of claim 9, comprising 0.85% (w/w) of powdered sugar substitute.

1 1. The composition of any one of claims 1 to 10, comprising from 0.12% to 0.15% (w/w) of powdered tartaric acid.

12. The composition of claim 1 1 , comprising 0.14% (w/w) powdered tartaric acid.

13. The composition of any one of claims 1 to 6, comprising an admixture of the following ingredients to make up a single dose of 175 mg:

145.25 - 150 mg powdered sugar substitute;

21 - 26.25 mg powdered tartaric acid; and

0.4 - 0.8 mg lyophilised VIP.

14. The composition of any one of claims 1 to 13, comprising or consisting of an admixture of the following ingredients:

powdered sugar substitute 0.85% (w/w);

powdered tartaric acid 0.14% (w/w); and

lyophilised VIP 0.0060% (w/w).

15. The composition of any one of claims 1 to 14, comprising or consisting of an admixture of the following ingredients:

powdered sugar substitute 149.4 mg;

powdered tartaric acid 25 mg; and

powdered VIP 0.6 mg.

16. The composition of any one of claims 1 to 15, comprising additional non-essential ingredients, including flavourants.

17. A method for manufacturing the composition of any one of claims 1 to 16, comprising or consisting of a step of admixing the powdered sugar substitute, powdered tartaric acid and lyophilised VIP with agitation.

18. The method of claim 17, further comprising a step of compression or compaction of the composition into a tablet form.

19. The method of claim 17 or 18, comprising the steps of first mixing the powdered sugar substitute and powdered tartaric acid by agitation, followed by addition of the lyophilised VIP with further agitation.

20. The method of claim 17 or 18, wherein the powdered sugar substitute, powdered tartaric acid and lyophilised VIP are mixed simultaneously with agitation.

21. A composition of any one of claims 1 to 20 for use in the management of inflammatory diseases or conditions in a subject. 22. A method of management of inflammatory diseases or conditions in a subject in need thereof by sublingual administration of the composition of any one of claims 1 to 20.

23. Use of VIP, a sugar substitute and tartaric acid of any one of claims 1 to 20 in the manufacture of a powder or tablet formulated for sublingual administration for management of inflammatory diseases or conditions in a subject in need thereof.

24. The composition of claim 21 , method of claim 22 or use of claim 23, wherein the inflammatory diseases or conditions are chronic or acute inflammatory conditions such as aches, pains, stiffness and minor joint swelling.

25. The composition of claim 21 or 24, method of claim 22 or 24 or use of claim 23 or 24, wherein the composition is administered to the subject as a dosage of 175 mg.

26. The composition, method or use of claim 25, wherein the composition is administered to the subject twice daily.