OPIOID PEPTIDE

Abstract

The present invention relates to an opioid peptide represented by general formula

TyrProPhe-X1-TrpGlyGly-X2-ProPro, wherein: X1 is represented by Gly or Lys; X2 is represented by Ile or Val.

The invention also relates to pharmaceutical, nutritional and nutraceutical compositions comprising the peptide and to the use of the same for analgesic purposes, and/or for providing a feeling of satiety, and/or for lowering arterial blood pressure in a subject.

Claims

1. Opioid peptide or salt thereof having naloxone-like binding activity characterized in that it is represented by formula:
TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein: X1 is represented by Gly or Lys; X2 is represented by Ile or Val.

2. A peptide according to claim 1 characterized by being SEQ D NO: 1.

3. A peptide according to claim 1 characterized by being SEQ D NO: 2.

4. A pharmaceutical composition characterized by comprising at least one opioid peptide having naloxone-like binding activity defined in claim 1 one of the or salt thereof and at least one pharmaceutically acceptable carrier.

5. A pharmaceutical composition according to claim 4, characterized by having a concentration of peptide or salt thereof in the composition ranging from 0.001% (w/w) 99.999% (w/w).

6. A pharmaceutical composition according to claim 4 characterized by comprising an analgesic compound.

7. A pharmaceutical composition according to claim 6 characterized by comprising morphine as the analgesic compound.

8. A pharmaceutical composition according to claim 4 characterized in that it is in the pharmaceutical form selected from a tablet, capsule, elixir, solution, suspension, emulsion, lotion, cream, ointment, granulate or powder.

9. A pharmaceutical composition characterized by comprising at least one pharmaceutically acceptable carrier and a promoter-forming agent of at least one peptide defined in claim 1.

10. A pharmaceutical composition according to claim 9 characterized by having an agent promoting a genetically modified microorganism.

11. A pharmaceutical composition according to claim 9 characterized in that it is in the pharmaceutical form selected from a tablet, capsule, elixir, solution, suspension, lotion, cream, ointment, granulate or powder.

12. A food composition characterized in that it comprises at least one food grade substance and at least one peptide defined in claim 1.

13. A food composition according to claim 12 characterized in that it comprises an amount of peptide or salt thereof ranging from 0.001% (w/w) to 99.999% (w/w).

14. A food composition characterized by comprising at least one food-grade substance and one promoter-forming agent of at least one peptide defined in claim 1.

15. A food composition according to claim 14 characterized in that it comprises as a promoter a genetically modified micro-organism.

16. A nutraceutical composition characterized in that it comprises at least one food grade substance and at least one peptide defined in claim 1.

17. A nutraceutical composition according to claim 16 characterized in that it comprises an amount of peptide or salt thereof ranging from 0.001% (w/w) to 99.999% (w/w).

18. A nutraceutical composition characterized in that it comprises at least one food-grade substance and one promoter-forming agent of at least one peptide defined in claim 1.

19. A nutraceutical composition according to claim 18 characterized in that it comprises as a promoter a genetically modified micro-organism.

20. The use of the peptide defined in claim 1 characterized in that it is for providing analgesia in a subject.

21. The use of the peptide defined in claim 1 characterized in that it is for providing a feeling of satiety to a subject.

22. Use of the peptide defined in claim 1 characterized in that it is for lowering arterial blood pressure in a subject.

23. The use of the pharmaceutical composition defined in claim 4, characterized in that it is for providing analgesia in a subject.

24. The use of the pharmaceutical composition defined in claim 4 characterized in that it is for providing a feeling of satiety to a subject.

25. The use of the pharmaceutical composition defined in claim 4 characterized in that it is for lowering arterial blood pressure in a subject.

26. The use of the food composition defined in claim 12 characterized in that it is for providing a feeling of satiety to a subject.

27. The use of the food composition defined in claim 12 characterized in that it is for lowering arterial blood pressure in a subject.

28. The use of the nutraceutical composition defined in claim 16 characterized in that it is providing a feeling of satiety to a subject.

29. The use of the nutraceutical composition defined in claim 16 characterized in that it is for lowering arterial blood pressure in a subject.

30. A method of activating the opioid receptor characterized in that it comprises administering to a subject an effective amount of at least one peptide defined in claim 1.

31. A method of activating the opioid receptor characterized in that it comprises administering to a subject a pharmaceutical composition as defined in claim 4.

32. A method of activating the opioid receptor characterized in that it comprises administering to a subject a pharmaceutical composition as defined in claim 12.

33. A method of activating the opioid receptor characterized in that it comprises administering to a subject a nutraceutical composition as defined in claim 16.

34. A method of providing analgesia characterized in that it comprises administering to a subject an effective amount of at least one peptide defined in claim 1.

35. A method of providing analgesia characterized in that it comprises administering to a subject a pharmaceutical composition as defined in claim 4.

36. A method of providing satiety to a subject characterized in that it comprises administering to a subject an effective amount of at least one peptide defined in claim 1.

37. A method of providing satiety to a subject characterized in that it comprises administering to a subject a pharmaceutical composition as defined in claim 4.

38. A method of providing satiety to a subject characterized in that it comprises administering to a subject a food composition as defined in claim 12.

39. A method of providing satiety to a subject characterized in that it comprises administering to a subject a nutraceutical composition as defined in claim 16.

40. A method of providing lowering of blood pressure characterized in that it comprises administering to a subject an effective amount of at least one peptide defined in claim 1.

41. A method of providing lowering of blood pressure characterized in that it comprises administering to a subject a pharmaceutical composition as defined in claim 4.

42. A method of providing lowering of blood pressure characterized in that it comprises administering to a subject a food composition as defined in claim 12.

43. A method of providing lowering of blood pressure characterized in that it comprises administering to a subject a nutraceutical composition as defined in claim 16.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0056] FIG. 1—Index of antinociception, in the tail flick test, represented by symbols (*, **, #) referring to a statistically significant activity (S) and or when significant differences do not occur (NS) when comparing SEQ ID NO:1 to saline, morphine and Leu-enkephalin. In (*) beginning of the activity of SEQ ID NO: 1 P<0.001 (S). (**) SEQ ID NO: 1×Morphine with P>0.05 (NS). In (*) Leu-enkephalin×SEQ ID NO: 1 P<0.05 (NS).

[0057] FIG. 2—Index of antinociception, in the tail flick test, referring to SEQ ID NO: 2 when compared to saline, morphine and Leu-enkephalin. In (*) beginning of the activity of SEQ ID NO: 2, P<0.001 (S). (**) SEQ ID NO: 2×Morphine with P>0.005 (NS). (#) Leu-enkephalin×SEQ ID NO: 2, P>0.05 (NS).

[0058] FIG. 3—Index of antinociception, in the hot plate, referring to SEQ ID NO: 1 when compared to saline, morphine and Leu-enkephalin. In (*) beginning of the activity of, P<0.01 (S). (**) SEQ ID NO: 1×Morphine with P>0.05 (NS). (#) Leu-enkephalin×SEQ ID NO: 1, P>0.05 (NS). (##) SEQ ID NO:1×SEQ ID NO:1+naloxone P>0.05 (NS).

[0059] FIG. 4—Index of antinociception, in the hot plate, referring to SEQ ID NO: 2 when compared to saline, morphine and Leu-enkephalin. In (*) beginning of the activity of SEQ ID NO: 2, P<0.001 (S). (**) SEQ ID NO: 2×Morphine with P>0.005 (NS). (#) Leu-enkephalin×SEQ ID NO: 2, P>0.05 (NS). (##) SEQ ID NO:2×SEQ ID NO:2+naloxone P>0.05 (NS).

[0060] FIG. 5—A. Determining the area below the curve in the tail flick test, for SEQ ID NO: 1; B. Determining the area below the curve in the tail flick test, for SEQ ID NO: 2

[0061] FIG. 6—A. Determining the area below the curve, in the hot plate test, for SEQ ID NO: 1; B. Determining the area below the curve, in the hot plate test, for SEQ ID NO: 2; B.

[0062] FIG. 7. Representation of the fragments obtained at the different incubation times with the enzyme trypsin. Analysis performed by mass spectrometry, MALDI. The first spectrum, marked with the letter A, refers to the time of 15 minutes; The second spectrum marked with the letter B indicates the time of 30 minutes; C indicates the time of 1 hour; D indicates the time of 2 hours; E indicates the time of 4 hours and, finally, F is the control peptide (SEQ ID NO: 2).

DETAILED DESCRIPTION OF THE INVENTION

[0063] The term “patient” used in this invention includes humans and also other mammals such as intensive and extensive breeding animals, zoo animals, companion animals such as cat, dog and horse.

[0064] The term “pharmaceutically acceptable carrier” used in the present invention refers to a diluent or adjuvant or excipient or carrier with which or in which the active compound is administered.

[0065] The term “therapeutically effective amount” used in the invention refers to the amount of active compound which, when administered to a patient to prevent or treat a condition (such as pain, hunger, etc.) is sufficient to effect such treatment. The therapeutically effective amount varies according to the active compound, the patient's condition, the severity of the condition, the age, weight and other characteristics of the patient.

[0066] The term “nutraceutical” used in the invention refers to a food or part of a food that provides medical and health benefits, including prevention and/or treatment of a condition.

[0067] The present invention relates to an opioid peptide or salt thereof having naloxone-like binding activity represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0068] X1 is represented by Gly or Lys;

[0069] X2 is represented by Ile or Val.

[0070] Preferably, the present invention relates to peptides or salts thereof having naloxone-like binding activity and sequences represented by SEQ ID NO: 1 and SEQ ID NO: 2.

[0071] The present invention further relates to a pharmaceutical composition comprising at least one opioid peptide or salt thereof having naloxone-like binding activity and at least one pharmaceutically acceptable carrier. The said peptide of the pharmaceutical composition is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0072] X1 is represented by Gly or Lys;

[0073] X2 is represented by Ile or Val.

[0074] Preferably, the peptides of the pharmaceutical composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0075] In the pharmaceutical composition of the invention the concentration of peptide or its salt having naloxone-like binding activity ranges preferably from 0.001% (w/w) to 99.999% (w/w).

[0076] In an alternative embodiment of the pharmaceutical composition of the invention, said composition further comprises an analgesic compound. Preferably, said analgesic compound is morphine.

[0077] The pharmaceutical composition of the invention may take various pharmaceutical forms. Among them, we can mention: tablet, capsule, elixir, solution, suspension, emulsion, lotion, cream, ointment, granulate, powder or lyophilized powder.

[0078] The present invention also relates to a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and a promoter-forming agent of at least one peptide having naloxone-like binding activity.

[0079] The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0080] X1 is represented by Gly or Lys;

[0081] X2 is represented by Ile or Val.

[0082] Preferably, the peptides of the pharmaceutical composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment of said pharmaceutical composition, the promoter is a genetically modified microorganism.

[0083] There are several pharmaceutical forms in which the pharmaceutical compositions of this invention may be present. Among these forms are the tablet, capsule, elixir, solution, suspension, lotion, cream, ointment, granulate, powder or lyophilized powder.

[0084] The present invention further relates to a food composition comprising at least one food grade substance and one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0085] X1 is represented by Gly or Lys;

[0086] X2 is represented by Ile or Val.

[0087] Preferably, the peptides of the food composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0088] In the food composition of the present invention the concentration of peptide or its salt having naloxone-like binding activity ranges preferably from 0.001% (w/w) to 99.999% (w/w).

[0089] The present invention also relates to a food composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0090] X1 is represented by Gly or Lys;

[0091] X2 is represented by Ile or Val.

[0092] Preferably, the peptides of the food composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment of said food composition of the invention, the promoter-forming agent is a genetically modified microorganism.

[0093] The present invention further relates to a nutraceutical composition comprising at least one food grade substance and one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0094] X1 is represented by Gly or Lys;

[0095] X2 is represented by Ile or Val.

[0096] Preferably, the peptides of the nutraceutical composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In the nutraceitucal composition of the present invention the concentration of peptide or its salt having naloxone-like binding activity ranges preferably from 0.001% (w/w) to 99.999% (w/w).

[0097] The present invention also relates to a nutraceitucal composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0098] X1 is represented by Gly or Lys;

[0099] X2 is represented by Ile or Val.

[0100] Preferably, the peptides of the nutraceutical composition of the present invention are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment of said nutraceitucal composition of the invention, the promoter-forming agent is a genetically modified microorganism. The present invention further relates to the use of the peptide defined in the invention in order to provide analgesia in an individual. The said peptide is an opioid peptide or salt thereof having naloxone-like binding activity represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0101] X1 is represented by Gly or Lys;

[0102] X2 is represented by Ile or Val.

[0103] Preferably, the invention relates to the use of at least one peptide selected from those having sequence represented by SEQ ID NO: 1 and SEQ ID NO: 2 for the purpose of providing analgesia in a subject.

[0104] The present invention further relates to the use of the peptide defined in the invention in order to provide a sense of satiety to a subject. The said peptide is an opioid peptide or salt thereof having naloxone-like binding activity represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0105] X1 is represented by Gly or Lys;

[0106] X2 is represented by Ile or Val.

[0107] Preferably, the invention relates to the use of at least one peptide selected from those having sequence represented by SEQ ID NO: 1 and SEQ ID NO: 2 for the purpose of providing a sense of satiety to a subject. The present invention further relates to the use of the peptide defined in the invention for the purpose of lowering blood pressure of a subject. The said peptide is an opioid peptide or salt thereof having naloxone-like binding activity represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0108] X1 is represented by Gly or Lys;

[0109] X2 is represented by Ile or Val.

[0110] Preferably, the invention relates to the use of at least one peptide selected from those having sequence represented by SEQ ID NO: 1 and SEQ ID NO: 2 for the purpose of lowering blood pressure of a subject.

[0111] The invention further relates to the use of the pharmaceutical compositions described in the invention for providing analgesia in a subject. Said analgesia occurs via the opioid receptor.

[0112] The invention further relates to the use of the pharmaceutical compositions described in the invention for lowering arterial blood pressure in a subject.

[0113] The invention further relates to the use of the pharmaceutical compositions described in the invention for providing a sense of satiety to a subject. Said sense of satiety occurs via the opioid receptor.

[0114] The invention further relates to the use of the food compositions described in the invention for providing a sense of satiety to a subject. Said sense of satiety occurs via the opioid receptor.

[0115] The invention further relates to the use of the food compositions described in the invention for lowering arterial blood pressure in a subject.

[0116] The invention further relates to the use of the nutraceutical compositions described in the invention for providing a sense of satiety to a subject. Said sense of satiety occurs via the opioid receptor.

[0117] The invention further relates to the use of the nutraceutical compositions described in the invention for lowering arterial blood pressure in a subject.

[0118] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject an effective amount of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0119] X1 is represented by Gly or Lys;

[0120] X2 is represented by Ile or Val.

[0121] Preferably, the peptides administered are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0122] Said administration of the opioid peptide of the invention may occur through external, enteral or parenteral application.

[0123] It is also an object of the invention a method of activating the opioid receptor, such method comprising administering to a subject a pharmaceutical composition comprising at least one opioid peptide or salt thereof having naloxone-like binding activity and at least one pharmaceutically acceptable carrier. The said peptide of the pharmaceutical composition is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0124] X1 is represented by Gly or Lys;

[0125] X2 is represented by Ile or Val.

[0126] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0127] Said administration of the pharmaceutical composition of the invention may be effected through external, enteral or parenteral application.

[0128] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and one promoter-forming agent of at least one peptide having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0129] X1 is represented by Gly or Lys;

[0130] X2 is represented by Ile or Val.

[0131] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. Also preferably, the promoter of the pharmaceutical composition is a genetically modified microorganism.

[0132] Said administration of the pharmaceutical composition of the invention may be effected through external, enteral or parenteral application.

[0133] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject a food composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0134] X1 is represented by Gly or Lys;

[0135] X2 is represented by Ile or Val.

[0136] Preferably, the peptides of the food composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0137] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject a food composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0138] X1 is represented by Gly or Lys;

[0139] X2 is represented by Ile or Val.

[0140] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0141] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0142] X1 is represented by Gly or Lys;

[0143] X2 is represented by Ile or Val.

[0144] Preferably, the peptides of the nutraceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0145] Another object of the invention is a method of activating the opioid receptor, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0146] X1 is represented by Gly or Lys;

[0147] X2 is represented by Ile or Val.

[0148] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0149] Another object of the invention is a method for providing analgesia, such method comprising administering to a subject an effective amount of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0150] X1 is represented by Gly or Lys;

[0151] X2 is represented by Ile or Val.

[0152] Preferably, the peptides administered are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0153] Said administration of the opioid peptide of the invention may be effected through external, enteral or parenteral application.

[0154] It is also an object of the invention a method of providing analgesia, such method comprising administering to a subject a pharmaceutical composition comprising at least one opioid peptide or salt thereof having naloxone-like binding activity and at least one pharmaceutically acceptable carrier. The said peptide of the pharmaceutical composition is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0155] X1 is represented by Gly or Lys;

[0156] X2 is represented by Ile or Val.

[0157] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0158] Said administration of the pharmaceutical composition of the invention may be effected through external, enteral or parenteral application.

[0159] Another object of the present invention is a method of providing analgesia, such method comprising administering to a subject a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and one promoter-forming agent of at least one peptide having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0160] X1 is represented by Gly or Lys;

[0161] X2 is represented by Ile or Val.

[0162] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. Also preferably, the promoter of the pharmaceutical composition is a genetically modified microorganism.

[0163] Said administration of the pharmaceutical composition of the invention may be effected through external, enteral or parenteral application. It is also an object of the invention a method of providing satiety to a subject, such method comprising administering to a subject an effective amount of at least one opioid peptide or salt thereof having naloxone-like binding activity.

[0164] The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0165] X1 is represented by Gly or Lys;

[0166] X2 is represented by Ile or Val.

[0167] Preferably, the peptides administered are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0168] Another object of the invention is a method of providing satiety to a subject, such method comprising administering to a subject a pharmaceutical composition comprising at least one opioid peptide or salt thereof having naloxone-like binding activity and at least one pharmaceutically acceptable carrier. The said peptide of the pharmaceutical composition is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0169] X1 is represented by Gly or Lys;

[0170] X2 is represented by Ile or Val.

[0171] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0172] It is also an object of the present invention a method of providing satiety to a subject, such method comprising administering to a subject a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and one promoter-forming agent of at least one peptide having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0173] X1 is represented by Gly or Lys;

[0174] X2 is represented by Ile or Val.

[0175] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. Also preferably, the promoter of the pharmaceutical composition is a genetically modified microorganism.

[0176] Another object of the invention is a method of providing satiety to a subject, such method comprising administering to a subject a food composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0177] X1 is represented by Gly or Lys;

[0178] X2 is represented by Ile or Val.

[0179] Preferably, the peptides of the food composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0180] Another object of the invention is a method of providing satiety to a subject, such method comprising administering to a subject a food composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0181] X1 is represented by Gly or Lys;

[0182] X2 is represented by Ile or Val.

[0183] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0184] Another object of the invention is a method of providing satiety to a subject, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0185] X1 is represented by Gly or Lys;

[0186] X2 is represented by Ile or Val.

[0187] Preferably, the peptides of the nutraceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0188] Another object of the invention is a method of providing satiety to a subject, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0189] X1 is represented by Gly or Lys;

[0190] X2 is represented by Ile or Val.

[0191] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0192] It is also an object of the invention a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject an effective amount of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0193] X1 is represented by Gly or Lys;

[0194] X2 is represented by Ile or Val.

[0195] Preferably, the peptides administered are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0196] Another object of the invention is a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a pharmaceutical composition comprising at least one opioid peptide or salt thereof having naloxone-like binding activity and at least one pharmaceutically acceptable carrier. The said peptide of the pharmaceutical composition is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0197] X1 is represented by Gly or Lys;

[0198] X2 is represented by Ile or Val.

[0199] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0200] It is also an object of the present invention a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and one promoter-forming agent of at least one peptide having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0201] X1 is represented by Gly or Lys;

[0202] X2 is represented by Ile or Val.

[0203] Preferably, the peptides of the pharmaceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. Also preferably, the promoter of the pharmaceutical composition is a genetically modified microorganism.

[0204] Another object of the invention is a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a food composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0205] X1 is represented by Gly or Lys;

[0206] X2 is represented by Ile or Val.

[0207] Preferably, the peptides of the food composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0208] Another object of the invention is a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a food composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0209] X1 is represented by Gly or Lys;

[0210] X2 is represented by Ile or Val.

[0211] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0212] Another object of the invention is a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and an opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0213] X1 is represented by Gly or Lys;

[0214] X2 is represented by Ile or Val.

[0215] Preferably, the peptides of the nutraceutical composition are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0216] Another object of the invention is a method of providing lowering of arterial blood pressure in a subject, such method comprising administering to a subject a nutraceutical composition comprising at least one food grade substance and a promoter-forming agent of at least one opioid peptide or salt thereof having naloxone-like binding activity. The said peptide is represented by the sequence:

TyrProPhe-X1-TrpGlyGly-X2-ProPro wherein:

[0217] X1 is represented by Gly or Lys;

[0218] X2 is represented by Ile or Val.

[0219] Preferably, the peptides are represented by SEQ ID NO: 1 and/or SEQ ID NO: 2. In a preferred embodiment, the peptide promoter-forming agent is a genetically modified microorganism.

[0220] Experiments and results obtained: Peptide synthesis: The synthesis of the peptides of the present invention was conducted using the Fmoc/t-butyl (9-fluorenylmethoxycarbonyl) strategy of manual synthesis in solid support (Chan & White, 2000) followed by purification by means of high performance liquid chromatography (HPLC) using a C18 reverse-phase semipreparative column (Vydac).

[0221] In vivo analyzes: Tests were carried out on Swiss mice of the Mus musculus species, in which nociception tests (hot plate and tail removal test) were used according to Le Bars et al. (2001). Injections were administered via intraperitoneal (IP). Doses were based upon dose and molaridade of the morphine (10 mg/kg animal). As a positive control, Leu-enkephalin (19.3 mg/kg animal) was used in addition to morphine. Sodium chloride saline was used as the negative control and, in order to suspend the antinociceptive effect of Leu-enkephalin and the peptide, serial doses (every hour) of Naloxone (4 mg/kg animal) were injected.

[0222] Digestion of the immobilized pepsin precursor: the peptide sequence SEQ ID NO: 2 was digested using immobilized pepsin for a period of 4 hours at 37° C. under agitation of 1400 RPM evaluating the fragmentation index at the times of 15 minutes, 30 minutes, 2 hours and 4 hours.

[0223] FIGS. 1,2,3 and 4 represent the raw data obtained from the experiments after statistical analysis. The symbols (*, **, #) indicate the beginning of the statistically significant activity (S) or the statistically non-significant differences (NS). Analyzes of opioid activity were performed in two classical models of studies for this activity: Tail Flick or Hot Plate test.

[0224] With the statistical analysis, it was possible to determine at which time (after application) the peptide started to present a statistically significant antinociceptive activity and compare it with the other groups in the tail-flick and hot plate tests.

[0225] In the tail-flick test, shown in FIG. 1, the peptide SEQ ID NO: 1 started its statistically significant action after 90 min. (*) up to 240 min. with P<0.001, in relation to the saline solution, negative control group. Comparing with morphine, positive control, SEQ ID NO: 1 showed no significant difference after 90 min. (**) with values of P>0.05. In the case of Leu-enkephalin, peptide positive control, there was no significant difference throughout the assay, i.e., SEQ ID NO: 1 showed a profile similar to that of Leukephalin (#) with P>0.05.

[0226] Except for the time of 30 min, there were significant differences between SEQ ID NO: 1 and SEQ ID NO: 1+naloxone during the 4 hour analysis, with P values <0.001.

[0227] As expected, there were no significant differences in activity profile between the following groups: saline, naloxone, SEQ ID NO: 1+naloxone and Leu-enkephalin+naloxone.

[0228] Analyzing the results for the peptide of SEQ ID NO: 2 (FIG. 2), its activity started at 90 minutes and lasted until the end of the test, except for the time of 150 minutes, when no significant difference was found in relation to the saline P<0.05. Regarding morphine, from the time of 120 minutes, there were no significant differences (P>0.005) with SEQ ID NO: 2 until the end of the test at 240 minutes. When compared to Leu-enkephalin, from the time of 60 minutes, SEQ ID NO: 2 did not show significant differences (#) with P>0.05.

[0229] At the other times (before 60 minutes) the difference was statistically significant with P<0.01. From 60 minutes there was a significant difference in the activity profile of SEQ ID NO: 2 when compared to SEQ ID NO: 2+naloxone with P<0.001 and it was maintained up to 240 minutes.

[0230] There were no significant differences in activity profile between the following groups: saline, naloxone, SEQ ID NO: 2+naloxone and Leu-enkephalin+naloxone.

[0231] In the Hot Plate test shown in FIG. 3, SEQ ID NO: 1 started its activity at 60 minutes, as it presented statistically significant differences in relation to saline with P<0.01 (*). It is noteworthy that at the time of 120 minutes, there was no significant difference, P>0.05.

[0232] Comparing the activity profile of the SEQ D NO: 1 with morphine, at the time of 30 minutes, there was a significant difference with P<0.001. From 60 minutes there were no statistically significant differences up to 240 minutes (**) with P>0.05. There were also no statistically significant differences between the SEQ ID NO: 1 and the Leu-enkephalin (#) with P>0.05, this value being constant throughout the test. In the assays with SEQ ID NO: 1+naloxone when compared to SEQ ID NO: 1, there were no significant differences only at 210 minutes and 240 minutes (##) P>0.05 which are the times at which the peptide returned to action after administration of naloxone was discontinued. With the discontinuation of naloxone administration in the naloxone+Leu enkephalin group, it can be observed that Leu-enkephalin activity has also been resumed.

[0233] There were no significant differences in activity profile between the following groups (which was expected): saline, naloxone, SEQ ID NO: 2+naloxone and Leu-enkephalin+naloxone.

[0234] Since the beginning of the test, the peptide SEQ ID NO: 2 (FIG. 4) started acting. There were statistically significant differences in relation to saline (*) with P<0.001 which persisted throughout the test. SEQ ID NO: 2 when compared to morphine (**) and Leu-enkephalin (#) did not show statistically significant differences, with P>0.05 during the entire assay.

[0235] At 180 minutes to 240 minutes there were no statistically significant differences between SEQ ID NO: 2+naloxone and SEQ ID NO: 2, (##) with P>0.05. The other times showed significant differences with P<0.001.

[0236] There were no significant differences in activity profile between the following groups: saline, naloxone, SEQ ID NO: 2+naloxone and Leu-enkephalin+naloxone.

[0237] Besides the analysis of variance, the area under the curve was also determined for each experimental group and positive controls (morphine and enkephalin). This analysis aimed at obtaining values for the cumulative effect acquired during the test. FIG. 5A shows that SEQ ID NO: 1, in the tail flick test, showed a cumulative effect over time greater than morphine and similar to Leu-enkephalin, which showed no statistically significant differences over the test with p>0.05 (NS).

[0238] In relation to the area below the curve, it can be noted that SEQ ID NO: 2 in the tail flick test showed lower cumulative effect to morphine and Leu-enkephalin (FIG. 5B).

[0239] The area below the curve on the hot plate, demonstrates that SEQ ID NO: 1 (FIG. 6A) exhibited a higher cumulative effect similar to morphine and Leu-encefaiina, because there was no significant difference (SEQ ID NO: 1×Leu-enkephalin) with P value>0.05 (NS) throughout the test.

[0240] The cumulative effect of SEQ ID No: 2 in the hot plate test (FIG. 6B) also showed indices area below the curve most similar to morphine and Leu-encefaiina (P>0.05).

[0241] Digestion of the precursor ion referring to SEQ ID NO: 2 (FIG. 7) shows that during incubation with trypsin enzyme (enzymes present in the gastrointestinal tract) the major peak of SEQ ID NO: 2 will reduce its intensity. Through this experiment it was possible to try to observe in vitro the digestion of the peptide.