WO2015104596A1

WO2015104596A1 - Vasoactive intestinal peptide

Info

Publication number: WO2015104596A1
Application number: PCT/IB2014/066544
Authority: WO
Inventors: Dr Hayley IPP
Original assignee: Ipp Dr Hayley
Priority date: 2014-01-10
Filing date: 2014-12-03
Publication date: 2015-07-16

Abstract

The invention discloses a vasoactive intestinal peptide (VIP) for switching off and/or preventing harmful and/or ongoing inflammation, which includes an amino acid sequence which is at least 90% SEQ ID NO: 1. The invention also extends to a pharmaceutical composition, which includes a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1, and being adapted to switch off and/or prevent hurtful and/or ongoing inflammation and/or treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndromein a subject.

Description

VASOACTIVE INTESTINAL PEPTIDE

FIELD OF INVENTION

The present invention relates to a vasoactive intestinal peptide.

More particularly, the present invention relates to a vasoactive intestinal peptide for the management and prevention of conditions associated with chronic inflammation.

BACKGROUND TO INVENTION

The social and economic impact of chronic disease is increasing globally as people are living longer. An important underlying cause of chronic disease is inflammation which has been linked strongly to the development of cancers and other aging-related conditions. Inflammation is a response to a perceived threat to the human body, whether it is in the form of an infection or injury. The body goes into 'fighting mode' in order to attack and clear the foreign or damaged area. If the infection or injury persists, the body continues to fight its presence, leading to potentially harmful inflammation. The fall-out of the ongoing 'fight mode' is damage to neighbouring healthy structures, such as the skin, arteries and immune system, causing accelerated aging, heart disease and cancers.

The body has natural mechanisms to limit the harmful effects of ongoing inflammation and will try to switch into the non-fighting or 'peaceful mode' in order to heal itself and prevent further damage.

The benefits of limiting ongoing inflammation are becoming increasingly evident. However, well-known anti-inflammatory agents have documented side-effects such as stomach ulcers and bleeding risks; while other agents such as cortisone, are known to suppress the immune system and are associated with increased risks of infection.

HIV infection is now well described as a chronic inflammatory condition with increasing risks of heart disease and cancers, despite effective treatment with antiretroviral (ARV therapy). In addition, there are multiple problems associated with ARV distribution particularly in resource-limited settings. Long term use of ARVs poses the risk of drug resistance, ARV associated toxicities and the persistence of chronic inflammation.

In South Africa, ARV therapy is commenced only when CD4 counts fall below a threshold of 350 cells/μΙ. However, during the stable chronic phase prior to the initiation of ARV therapy, extensive immune-pathological damage occurs. Aberrant immune activation (in multiple cell types involved in the immune response), chronic inflammation and apoptosis (particularly within the CD4 lymphocyte subset) are the hallmarks of this phase of infection. By reducing cell loss and dysfunction resulting from these processes, one could extend the period prior to access to ARVs and also reduce detrimental effects associated with chronic infection.

There is thus an urgent need for novel drug targets or treatment approaches for HIV worldwide, particularly in countries where there is a high disease burden, patient access to treatment is difficult and complications with existing drug therapy regimens remain problematic.

In developing countries, the treatment should preferably not only be effective, but also affordable and easily administered.

It is an object of the invention to suggest a vasoactive intestinal peptide to assist in overcoming the aforementioned problems. SUMMARY OF INVENTION

According to the invention, a vasoactive intestinal peptide (VIP) for switching off and/or preventing ongoing inflammation system includes an amino acid sequence which is at least 90% SEQ ID NO: 1 .

Also according to the invention, there is provided the use of a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 , in the manufacture of a medical composition for switching off and/or preventing ongoing inflammation and/or treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome in a subject.

Yet further according to the invention, there is provided a pharmaceutical composition, include a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 for supporting the immune system and/or treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome in a subject.

Yet further according to the invention, there is provided a method of switching off and/or preventing harmful and/or ongoing inflammation and/or treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome, in a subject, the method including the step of administering a therapeutically effective amount of a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 .

The amino acid sequence may be 95% SEQ ID NO: 1 . The amino acid sequence may be 100% identical to SEQ ID NO: 1 .

The medical composition may further include a pharmaceutically acceptable carrier.

The medical composition may further include a taste enhancer.

The medical composition may further include an anti-oxidant.

The vasoactive intestinal peptide (VIP) may be used in treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome.

The vasoactive intestinal peptide (VIP) may be used to support, to treat and/or modulate the immune system.

The vasoactive intestinal peptide (VIP) may be used for treating chronic inflammations and/or disorders.

The medical composition may be formulated for inhalation, subcutaneous, intranasal, intravenous, sublingual, buccal, rectal and/or oral administration.

The binding of the VIP to its receptors on CD4 immune cells may result in the triggering of an inhibitory pathway in T cells.

The subject may be preferably a human.

The subject may have newly been diagnosed HIV infection or chronic HIV infection. BRIEF DESCRIPTION OF DRAWINGS

The invention will now be described by way of example with reference to the accompanying schematic drawings. In the drawings there is shown in:

Figure 1 : a graph showing that levels of expression of the receptor for VIP; VPAC2, were significantly increased in HIV infection compared with uninfected persons;

Figure 2: a graph showing the reduction of activation-induced expression of Fas

Ligand by VIP; and

Figure 3: a graph showing that CD4 T cells in HIV infection retain the ability to respond to stimulation via their T cell receptor for antigen (sCD3) by maintaining the ability to express CD25 (the receptor for the growth factor interleukin-2).

DETAILED DESCRIPTION OF DRAWINGS

The use of vasoactive intestinal peptide (VIP) for treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and chronic inflammatory response syndrome, a subject is described herein.

According to the invention, a vasoactive intestinal peptide (VIP) for switching off and preventing harmful and ongoing inflammation includes an amino acid sequence which is at least 90% or 95% SEQ ID NO: 1 . or is even 100% identical to SEQ ID NO: 1 .

The vasoactive intestinal peptide (VIP) can be used in the manufacture of a medical composition for switching off and preventing harmful and ongoing inflammation and treating HIV infection, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and chronic inflammatory response syndrome, in a subject. The invention also discloses a method of switching off and preventing harmful and ongoing inflammation and treating HIV infection, arthritis and rheumatoid arthritis, bio- toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome in a subject, the method including the step of administering a therapeutically effective amount of a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% or 95% SEQ ID NO: 1 . or is even 100% identical to SEQ ID NO: 1 .

The medical composition further includes a pharmaceutically acceptable carrier. The medical composition includes a taste enhancer. The medical composition includes an anti-oxidant.

The vasoactive intestinal peptide (VIP) is used in treating HIV infection, arthritis and rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome,.

The vasoactive intestinal peptide (VIP) can be used to support, to treat and/or modulate the immune system.

The vasoactive intestinal peptide (VIP) can be used for treating chronic inflammations and/or disorders.

The medical composition can formulated for inhalation, subcutaneous, intranasal, intravenous, sublingual, buccal, rectal and/or oral administration. The binding of the VIP to its receptors on CD4 immune cells results in the triggering of an inhibitory pathway in T cells. VIP has been shown to decrease the expression of the death signaling molecule Fas Ligand.

Thus the above represents a novel approach to dampen the effects of activation in a previously activated compartment of CD4 T cells and limit further cell death.

The subject is preferably a human and may have newly been diagnosed HIV infection or chronic HIV infection.

Contrary to the previously accepted model of HIV being primarily an immunosuppressive disease, it is now becoming evident that this disease may in fact be driven by aberrant immune activation. The extent of CD4 T cell activation is the most accurate predictor of disease outcome. The result of continued immune activation is depletion of immune cells (especially CD4 cells) and immune exhaustion (loss of function in many immune cell subsets). Immunotherapeutic approaches attempting to "restore" immune function or assist dysfunctional T cells, e.g. via IL-2 therapy, are flawed in that they fuel the activation cascade. On the other hand, extensive blanket immunosuppression, e.g. by administration of cyclosporine A, predisposes HIV-infected patients to the risk of other opportunistic infections. Vaccination also poses risks, in that the induction of an immune response via vaccination may make the individual more susceptible to infection as more activated cells are made available for viral targeting and infection.

The approach of this invention is therefore to 'switch off T cells selectively; whilst maintaining their ability to respond at another stage to foreign antigens. Activation of the immune system in the acute stage of HIV infection is critical for the initial containment of the virus. However, it is this activation process that paradoxically becomes harmful and promotes progression of the disease. Activated CD4+ T cells are the preferred targets for viral infection and replication and, therefore, the persistent activation of the immune system aggravates the problem. In addition, repeated activation of the CD4+ T cells promotes the up-regulated expression of Fas ligand which is the death-signal for any neighbouring cell expressing the receptor Fas.

In HIV infection, the level of immune cell activation is strongly correlated with progression of the disease, and is a more accurate predictor of adverse outcome than either low CD4 count or high viral load. Several approaches have been attempted to 'dampen' this activated state associated with HIV infection, with varying degrees of success. General immune-suppressants such as hydroxyl-urea, prednisone and mycophenolic acid have shown limited benefit or even harm in some cases, illustrating that blanket immunosuppression is counterproductive. A randomized trial which added cyclosporin A to a HAART regimen in the chronic stage of the disease showed only minimal and transient increases in CD4+ T cell counts.

The neuropeptide vasoactive intestinal peptide (VIP) is a 28 amino acid peptide member of the secretin-glucagon family of peptide hormones (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 ). Its effects on the immune system are predominantly inhibitory, mediated via the adenyl cyclase/cyclic AMP second messenger system. VIP mediates these effects via two homologous, G-protein- coupled receptors designated VPAC1 and VPAC2, found on several cell types including T cells and other immune cells such as dendritic cells. These receptors are variably expressed in CD4+ T cells, depending on the activation state of the cell: VPAC1 is the predominant receptor present in the resting state, whilst VPAC2 is up- regulated on activation of the cell. The differential expression of VPAC1 and 2 is related to activation status of T cells and the differential outcome of VIP interaction with VPAC1 (pro-inflammatory) and VPAC2 (anti-inflammatory) has been described in various animal models of disease.

VIP has been investigated previously in the context of autoimmune conditions, such as rheumatoid arthritis where the body attacks itself, in lung conditions requiring inhalation of the peptide and in sepsis which is an acute, life-threatening condition requiring intravenous administration for rapid effects.

This is the first known description of the use of VIP for switching off and preventing harmful and ongoing inflammation and treating HIV infection.

The VIP can be incorporated into a composition with a pharmaceutically acceptable carrier.

The VIP can be incorporated into a composition with a taste enhancer and/or an antioxidant.

The composition may be formulated for subcutaneous, sublingual, oral, intra-nasal, or intravenous administration, or any other suitable form of administration. In particular, where the composition is intended for use in developing countries where there is limited access to medical expertise, the preferred administration route will be oral or sublingual or buccal administration. Administration of the composition would selectively inhibit CD4+ T cell death signals, whilst maintaining the ability of the cells to be activated in response to antigen via the T cell receptor. This would prevent T cell loss and/or delay immune exhaustion and progression to AIDS.

The applicant anticipates that targeting VPAC2 with VIP will allow for induction of VIP- mediated inhibitory effects in cells expressing this receptor.

The present invention is further described by the following examples. These examples, however, are not to be construed as limiting in any way either the spirit or scope of the invention.

Example 1

87 untreated HIV-infected individuals with CD4 counts >200, and 57 uninfected controls were recruited from a primary health clinic in Cape Town, South Africa. Flow Cytometry was used to determine levels of expression of VPAC1 and VPAC2, as well as FasL on CD4+ T cells and these results were correlated with the immune activation phenotype %CD38+CD8+ T cells. VPAC2 expression was significantly increased in the HIV group (mean %VPAC2+ CD4+ cells 19.25 vs. control 12.56; P=<0.0001 ) as shown in Figure 1 , but no difference in VPAC1 expression was observed. VPAC2 correlated positively with FasL (r= 0.310; p=0.001 ) and there was a significant inverse correlation between FasL and the CD4 count (r= -0.21 1 ; p=0.013) and a direct correlation with %CD38+CD8+ T cells (r=0.39; p=<0.0001 ). Thus higher levels of immune activation correlated with higher levels of the death signaling FasL and lower CD4 counts. VPAC2 may provide a novel target for the selective limitation of CD4+ T cell death in HIV infection. Example 2

Fas Ligand expression was measured on CD4+ T lymphocytes in a whole blood assay at baseline and after 21 hours of stimulation with anti-CD3 monoclonal antibody (which mimics stimulation of the CD4 T cell receptor for antigen), with and without VIP at 10^"3 M.

VIP was shown to reduce activation-induced expression of Fas Ligand from a mean of 44.46% to 30.18%. Baseline levels without stimulation were 21 .59%; therefore VIP inhibited activation-induced FasL expression by 57% (see Figure 2). VIP may therefore limit FasL-induced death signaling in CD4+ T cells and thereby prevent further decline in CD4 count and the development of AIDS.

Example 3

The receptor on CD4+ T cells for the T cell growth factor, lnterleukin-2 is CD25. Levels of CD25 expression were measure on CD4+ T lymphocytes in a whole blood assay at baseline and after 21 hours of stimulation with anti-CD3 monoclonal antibody, with and without VIP at 10^"3 M.

Incubation with VIP did not limit the CD4 T cells' ability to up-regulate the expression of CD25 (see Figure 3); therefore providing evidence of a retained functional response to T cell receptor stimulation. In other words, VIP does not induce generalized immunosuppression and thus may be particularly useful in the context of the management of HIV infection. Conclusion

The pharmaceutical composition according to the invention, which includes a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 for supporting the immune system and/or treating HIV infection in a subject, may be delivered by means of nasal sprays, aerosol inhalants, intravenous route and/or orally. In the latter the pharmaceutical composition may be in a dissolvable format with minimal gastrointestinal breakdown or 'first-pass' effect. Surface linings without skin coverings; such as in the mouth, are rich in blood vessel supply and would provide the optimal surface environment for direct absorption into the blood stream without significant breakdown of the substance. Therefore, drug delivery systems utilizing the oral (buccal or sublingual) or rectal mucosae routes is important.

Claims

PATENT CLAIMS

1 . A vasoactive intestinal peptide (VIP) for switching off and/or preventing harmful and/or ongoing inflammation, which includes an amino acid sequence which is at least 90% SEQ ID NO: 1 .

2. A peptide as claimed in claim 1 , in which the amino acid sequence is 95% SEQ ID NO: 1 .

3. A peptide as claimed in claim 1 , I which the amino acid sequence is 100% identical to SEQ ID NO: 1 .

4. A peptide as claimed in any one of the preceding claims, which is adapted to be used in treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome.

5. A peptide as claimed in any one of the preceding claims, which is adapted to be used to support, to treat and/or modulate the immune system.

6. A peptide as claimed in any one of the preceding claims, which is adapted to be used for treating chronic inflammations and/or disorders.

7. A peptide as claimed in any one of the preceding claims, in which binding of the peptide to its receptors on CD4 immune cells results in the triggering of an inhibitory pathway in T cells.

8. A use of a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 , in the manufacture of a medical composition for supporting the immune system and/or treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndromein a subject.

9. A use as claimed in claim 8, in which the medical composition further includes a pharmaceutically acceptable carrier.

10. A use as claimed in claim 8 or claim 9, which the medical composition is formulated for inhalation, subcutaneous, intranasal, intravenous, sublingual, buccal, rectal and/or oral administration.

1 1 . A use as claimed in any one of claims 8 to 10, in which the subject is preferably a human.

12. A use as claimed in any one of claims 8 to 1 1 , in which the subject has newly been diagnosed with HIV infection or chronic HIV infection.

13. A pharmaceutical composition, which includes a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 , and being adapted to switch off and/or prevent hurtful and/or ongoing inflammation and/or treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndromein a subject.

14. A composition as claimed in claim 13, which includes a pharmaceutically acceptable carrier.

15. A composition as claimed in claim 13 or claim 14, which is formulated for inhalation, subcutaneous, intranasal, intravenous, sublingual, buccal, rectal and/or oral administration.

16. A composition as claimed in any one of claims 13 to 15, in which the subject is preferably a human.

17. A composition as claimed in any one of claims 13 to 16, in which the subject has newly been diagnosed HIV infection or chronic HIV infection.

18. A method of switching off and/or preventing hurtful and/or ongoing inflammation and/or treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndromein a subject, which includes the step of administering a therapeutically effective amount of a vasoactive intestinal peptide (VIP) having an amino acid sequence which is at least 90% SEQ ID NO: 1 .

19. A vasoactive intestinal peptide (VIP) for limiting the harmful effects of inflammation and/or switching off and/or preventing hurtful and/or ongoing inflammation substantially as hereinbefore described with reference to the accompanying drawings.

20. A use of a vasoactive intestinal peptide (VIP) substantially as hereinbefore described with reference to the accompanying drawings.

21 . A pharmaceutical composition substantially as hereinbefore described with reference to the accompanying drawings.

22. A method of limiting the harmful effects of inflammation and/or switching off and/or preventing hurtful and/or ongoing inflammation and/or treating HIV infection, arthritis, rheumatoid arthritis, bio-toxin related illnesses, fibromyalgia, chronic fatigue, and/or chronic inflammatory response syndrome, substantially as hereinbefore described with reference to the accompanying drawings.