IL92996A - Photoactive compounds as medicaments for the treatment of virus infections and method for production of vaccines - Google Patents

Description

E OF PHOTOACTIVE COMPOUNDS AS MEDICAMENTS FOR THE TREATMENT OF VIRUS INFECTIONS AND METHOD FOR PRODUCTION OF VACCINES USE OF PHOTOACTIVE COMPOUNDS AS MEDICAMENTS FOR THE TREATMENT OF VIRUS INFECTIONS AND METHOD FOR PRODUCTION OF VACCI ES FIELD OF THE INVENTION The present invention relates to the field of particularly to the use of certain photoactive compounds in the inactivation attenuation of viruses virus infected particularly virus infected via and the use of said inactivated attenuated viruses virus infected cells to engender an immune A particular aspect of the invention relates to the use of said photoactive compounds as medicaments for the treatment of patients who are infected with a particularly an HIV and who have an abnormally low white blood cell by using photopheresis in combination with the administration of a photoactive compound such as The invention also relates to vaccines against particularly HIV and methods for producing said BACKGROUND INVENTION debilitating infectious diseases which attributed to viral There are also different types of viruses including DNA viruses and RNA Retroviruses form a of RNA viruses in order must first employ reverse transcription of the RNA of their genome into DNA describes tha synthesis of RNA from Once in the of viral is incorporated into the host cell it to take full advantage of the host on machinery for the purpose of once incorporated into the host the virus persist for as long as the Certain retroviruses are known to cause a depression in an white blood cell This of which reduce white blood call count are known as Particular species of HIV retroviruses have been isolated from patients who suffer Acquired Immune Deficiency Syndrome and have been given the designations HIV 1 and HIV sometimes collectively referred to as or simply These retroviruses will infect cells expressing the such as human and These involved in functioning of the infect in results in the progressive loss of the cell population and disturbs the function of other CD4 such thereby reducing ability to other and predisposing patient to infections which frequently prove There are at least three clinical manifestations of AIDS the initial the only indication of infection is either the presence of antibodies in the or the ability to culture the The next known as related and the symptoms associated therewith may include persistent gene l general increased temperature and This condition usually progresses to the fatal AIDS when the patient loses the ability to A particularly troublesome problem associated with combatting HIV retrovirus infections is that the to reverse transcription process is fraught with repeated mutation which makes it extremely difficult for immune system to recognize and attack infected cells along with the virus It is an object of this invention to provide a method of treating viru3 infections in patients having an abnormally low white blood cell It is a further abject of the invention to provide a of treating patients with an HIV A still further object of the invention is to provide a vaccine against virus particularly when the infection by retrovirus such an Other objects of the invention will be apparent from the of invention which This invention relates to the use of photoactive compounds as a medicament for the treatment of viral infections associated with abnormally low counts of lymphocytes or white blood In accordance with the present invention a method has f for treating who infected with a an HIV using a photoactive compound that b in th case of a virus to cell membrane by binding to a receptor a nucleic acid on cell to nucleic acid in the cell nucleus or cell in case of either virus or associated that binds to the virus surface to a to a nucleic fragment on tha virus to nucleic acid DNA or which is incorporated in the upon activation by exposure to electromagnetic radiation of a prescribed such as ultraviolet for the purpose of inactivating attenuating the virus and permitting the so treated virus virus cells to presented to the immune of the for the purpose of engendering an immune response to the viral Psoralen compounds are particularly preferred for this especially the compound in which case UVA radiation is preferred for activating said EP teaches the use of f rocoumarins in the decontamination of blood by inactivation of polynucleotides such decontaminated blood being intended for use in uninfected recipients EP discloses a method for the inactivation of Human Tumor Leukemia Virus while maintaining its for use in in vitro diagnostic US discloses a method of reducing a lymphocyte count by in vitro treatment of the blood with psoralen and ultraviolet and US describes vaccines against viruses obtained from normal or tumor In contrast to the present invention relates to the use of a substance as a medicament for the treatment of a virus in an infected or for producing a vaccine to a An especially important feature of the present invention is that the treatment methods described herein are used in patients having an abnormally low number of circulating white blood In it has been found by that the treatment methods according to the invention can be used to reconstitute the immune in such This is of tremendous importance in the treatment of patients in addition to having an HIV infection itself heretofore been extremely difficult to also have a depressed immune Such patients generally cannot tolerate a reduction in the number of their Lymphocytes or white blood cells without succumbing to opportunistic has thus been found by the inventors that the treatment methods in accordance with the present invention useful to control and other retrovirus infections in patients having an abnormally low white blood cell without causing a harmful depression of the immune the methods of the invention can employed in the of conditions such as without subjecting the treated patient to a of opportunistic The treatment methods in accordance with the invention may also be used in such patients to improve the functioning of their immune syste In HIV retroviruses may also be responsible for causing diseases other than The inventors believe that the inventive methods should be useful for treating such other diseases as In accordance with the a photoactive compound such is administered to the or fraction in vitro or in vivo using administration rou A portion of a blood then using photop which comprises blood to electromagnetic radiation in a wavelength suitable for activating such as ultraviolet preferably long wavelength ultraviolet light in the wavelength range of 320 to 400 commonly called UVA treated or a fraction returned to the patient the case of or remains in patient in vivo vaccines against and methods of making are also pxovided according to the According to the a photoactive compound described is administered to the blood or some fraction thereof of a donor who is infected with a such an HIV retrovirus who is suffering from AIDS or AIDS Related At least a portior of the blood is then treated using as described The treated blood or some fraction thereof treated free isolated may be used as a In the case of treating an HIV infection it is preferred to use treated virus infected cells along with the virus in order to obtain the desired immune the treated blood is processed by conventional techniques to the The resulting processed fraction is then used as which can be to a 1 is a block of a apparatus which can used to practice the invantiva a graph of changes in CD and GP24 and GP120 levels of patient 3 in Example is a graph of in antibody levels of the five treated in Example 4 a graph of changes in antibody levels of the five patients treated in Example and 5 is a graph of changes in CD helper cell percentages of five patients treated in the according to the invention autogenous vaccine againet viral inventive methods have been found by the inventors to be useful in the treatment of patients having a virus infection and who have an abnormally low white blood cell count and particullarly useful in treating HIV retrovirus The inventive methods also particullarly useful for treating patients who are AIDS Carriers or who have AIDS or AIDS Related According to the claimed photoactive compound is first administered to the blood of a patient who is infected with a The photoactive compound may be administered in vivo orally or or may administered in vitro to a portion of the blood which has removed from the patient by employing conventional blood withdrawal free virus is isolated from infected calls using conventional virus isolation methods which are known in photoactive compound can be administered to the infected cells prior to virus isolation or can be administered to the free isolated the case of treating HIV it is presently preferred to use both treated virus and treated virus infected cells in the methods described In accordance with the present the compound selected should preferably be one that in case of a virus infected to the cell by binding to a receptor a nucleic acid fragment on the to nucleic in the cell nucleus or cell in the case of either free virus or cell associated that binds to the virus surface to a receptor to nucleic acid fragment on the virus to nucleic acid or which is incorporated in upon activation by exposure to electromagnetic radiation of a prescribed such as ultraviolet for the purpose of inactivating attenuating the virus and permitting the so treated virus virus infected cells to be presented to the immune system of the Psoralen compounds are particularly preferred for this especially the compound in which case UVA radiation is preferred for activating said the portion of the or the free isolated to which the photoactive compound has been administered is treated by subjecting the portion of the or the free isolated to photopheresis using said radiation for ultraviolet The photop step is preferably carried out in vitro using an extracorporeal The photopheresis step in accordance with the present invention may also be carried out in Ά presently preferred extracorporeal photopheresis apparatus for use in the methods according to the invention currently manufactured by under the name description of the Therakoe apparatus nay be found in Patent granted to Edelson on August the contents of which are hereby incorporated by reference in their As illustrated diagramatically in the apparatus includes a pump 10 for removing blood the patient via a donor placed in an appropriate vein of the an irradiation chamber a radiation source 30 in close proximity to the Irradiation chamber and a centrifuge preferably of the continuous The various parts of the such as tu ing Collection bags for the blood and the which come in contact with blood or some fraction are preferably replaceable so that they may be disposed of after each use to prevent the possibility of transmitting infections from one patient to others who are subsequently treated with the The exposure of or free isolated to let light in a photopheresis apparatus is within the ability of persona having ordinary skill in the When the photopheresis step is carried out in at least a fraction of the treated or the treated free isolated is returned to the patient following the photopheresis the treatment method hereinabove is repeated at an interval of about once per to about once four Most in the the treatment methods described herein are on two successive days and repeated approximately once per month the patient preferably receives two treatments every In view of the disclosure contained those who are in the art will be able to adjust the treatment parameters dosage of the photoactive compound and periodicity of treatment and the number of treatments administered in each period twice per month on two successive depending on the condition of the patient and the response to the Preferred photoactive compounds for use in accordance with the present invention are compounds known as psoralens which are described in Patent the disclosure of which is incorporated herein by reference in their The preferred photoactive compounds for use in accordance with the present invention include the psoralen treatment according to the is preferably carried out using long wavelength ultraviolet light at a wavelength within the range of 320 to 400 The exposure to ultraviolet light during the treatment preferably a duration of about three to four although shorter or longer periods may used if Whatever the spectrum of electromagnetic the exposure of virus infected cells virus following administration of the photoactive should of sufficient to effectively inactivate the The selection of an appropriate wavelength for as well as the depending upon the compound being employed and tha conditions of in vivo exposure or in vitro the ability of those skilled in the art in view of the presort When the photoactive compound is it is in accordance with invention to utilize an exposure to UVA radiation of about 2 based upon the surface area of the virus and virus infected cells undergoing When the photopheresis treatment according to the invention is carried out in careful attention should be paid controlling the maximum radiant exposure so as to avoid ry o radiant exposure to ultraviolet light known in the shall not be described herei In the invention provided novel treatment for patients who are infected by a virus and who have depressed immune systems as a result of such as well as for who are infected with an HIV retrovirus or who are AIDS Carriers or who have AIDS or S Related Such patients cannot tolerate a treatment that would depress their immune The treatment methods according to the invention have been by the inventors to be safe in this latter regard while also being effective in combatting HIV infection in The invention also provides methods for making According to the a donor who is infected with such as an HIV may be utilized to produce a vaccine against his infection as a photoactive compound as described hereinabove is administered to at least a portion of the blood conta free virus virus infected cells either prior to of the either orally or or after removal from the donor in which case it is administered in a portion of the blood could first be processed using known methods to substantially remove the erythrocytes and the photoactive compound is then administered to resulting fraction isolated to which the photoactive compound been administered is subjected to a treatment using radiation of a prescribed preferably in the previously The treated the treated portion thereof or the treated free isolated virus the case may then administered back to the donor as an autogenous vaccineJ It will be understood that in accordance with the present invention the treated virus can also be isolated from the treated blood or portion thereof following photopheresia treatment for use as a in accordance with present the treated which is itself a mixture of various blood components including peptides or the treated portion blood may ba as is within the ability of laving ordinary skill in the to isolate a particular or components which may be used in the treatment of the virus inflection of the donor may be used as a vaccine against Passages of the specification which are not within the scope of the claims do not constitute a part of the 1 A male 39 years of weighing approximately 70 kg who had been diagnosed as having AIDS Related Complex was treated in accordance with the present invention as was administered orally to the Inclusion criteria included only patients not having undergone any previous therapy against HIV Each patient received a monthly regimen of the substantially described in Example repeated on two The dosage of employed varied between and and was administered Four of the patients received a total of twelve each over the six course of the that on successive days at intervals for six The fifth patient voluntarily discontinued treatment after the fifth month and only received ten On the basis of the physical monthly antibody and antigen and all five patients had either a stabilization of theLr disease or a positive response to the None of the patients experienced damage to their immune systems during the of the By strictly clinical all five patients demonstrated improvement with respect to lymph node Energy index general state of improved in At the start of the increase in energy level lasted for approximately one By third month of a more consistent rise in energy level Most the first patient treated patient in Example a homosexual male who before treatment was able to walk up a flight of stairs only with is now capable of jogging three and miles per day and of er ara e a rs remained unchanged in all Those patients who antigen negative remained those who were positive and no changes in titer occured by the of the viral cultures of blood for HIV turned negative in one patient but remained positive in the others afte the reported The course of the patient whose HIV culture turned negative is outlined in The Gp 24 levels of all five patients over the course of treatment are in The Gp 120 antibody levels off all five over the course of treatment are plotted in the helper cell percentages of all five patients over course of are plotted in I A summary of the clinical characteristics and laboratory results is provided in Table 2 s herein bo by pereona having ordinary skill in tha art departing from spirit and scope thereof aa defined in to lowing insufficientOCRQuality

Claims (21)

92996/3 WE CLAIM:

1. A photoactive compound, namely psoralen, psoralen derivatives or combinations of psoralen and/or psoralen derivatives, for use as a medicament for the treatment of a virus infection associated with an abnormally low number of lymphocytes or white blood cells.

2. A compound according to claim 1, which is a compound that binds to the cell membrane and/or to nucleic acid in the cell nucleus or cell cytoplasm.

3. A compound according to claim 1, which is a compound that binds to the virus surface and/or to nucleic acid which is incorporated in the virus.

4. A compound according to claim 1, which is reactive to ultraviolet light.

5. A compound according to any one of claims 1 to 4, which is a psoralen compound.

6. ^6. A compound according to claim 5, which is selected from the group consisting of psoralen, 8-methoxypsoralen, 4, 5 '-8- trimethylpsoralen, 5-methoxypsoralen, 4-methylpsoralen, 4, 4-dimethylpsoralen, 4, 5 ' -dimethylpsoralen, and 4', 8- dimethylpsorale . 92996/3

7. A compound according to claim 5, which is selected from the group consisting of 8-methoxypsoralen, psoralen and 4, 5', 8-trimethylpsoralen .

8. A compound according to claim 5, wherein the psoralen compound is 8-methoxypsoralen.

9. A compound according to any one of claims 1 to 8, for use as a medicament for the treatment of AIDS or AIDS Related Complex.

10. A compound according to any one of claims 1 to 8, for use as a medicament for the treatment of an HIV Retrovirus infection.

11. A pharmaceutical preparation comprising an effective amount of a compound according to any one of claims 1 to 10.

12. A pharmaceutical preparation according to Claim 11 in intravenously administrable form.

13. A pharmaceutical preparation according to claim 11, in injectable form.

14. A pharmaceutical preparation according to claim 11, in orally administrable form.

15. Use of a compound according to any one of claims 1 to 7, for the manufacturing of a medicament for the treatment of viral infections, as described in the specification. 3i 92996/3

16. A method Cor producing a vaccina against an HIV retrovirus infection in an infected donor, said method comprising the steps of; a. administering in vitro to at least a portion of the donor's blood a photoactive compound that binds, in the case of a virus infected cell, to the cell membrane (e.g., by binding to a receptor and/or a nucleic acid fragment on the cell membrane) and/or to nucleic acid in the cell nucleus or cell cytoplasm, or, in the: case of either free virus or cell associated virus, that binds to the virus surface (e.g., to a receptor and/or to a nucleic acid fragment on the virus surface) and/or to nucleic acid ([e.g., DNA or RNA) which is incorporated in the virus, upon activat ion by exposure to electromagnetic radiation of a prescribed spectrum for the purpose of inactivating and/or attenuating the virus and permitting the so treated virus and/or virus J.nfected cells to be presented to the immune system of the patient for the purpose of engendering an immune response to the viral .nfectionj and b. then treating the portion o£ the donor's blood to which :he photoactive compound has been administered, said treatment comprising subjecting at least a fraction of said portion of blood to photopheresis using said electro-magnetic radiatLon, said photoactive compound and said electro-magnetic radiatLon being administered in amounts which are 92996/3 pharmaceutically effective for attenuating and/or inactivating at least some of the HIV retrovirus which is present in the treated fractipn.

17. The method of claim 16. wherein the fraction of blood |:Ls processed to substantially remove erythroc tes.

18. The method of claim 17, wherein the fraction of blood lie processed to substantially remove erythrocytes prior to perfoijming step (b) .

19. The method of claim 17. wherein the fraction of blood is processed to .substantially remove erythrocytes after perfoifming step (b) .

20. The method of claim 16. wherein the fraction which is treated in step (b) consists essentially of free isolated HIV retro irus .

21. The method of claim 16. further .comprising procensing the treated fraction of blood following step (b) to isolate treated HIV retrovirus for use s a vaccine against an HIV r«troviru« infection.