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Aspidosperma subincanum II. Usefulness of uleine and ribonucleic fragments in the treatment of AIDS patients

Abstract

Aids patients were treated during a year with three different food supplements commercially available: para-pau-aspido (Aspidosperma subincanum Mart. ex A. DC., Apocynaceae); 2Leid (nucleic acids and cytokines); and Para Immuno (propolis, pollen and royal jelly). All foods, given either alone or in combination, proved useful to all AIDS patients who received the supplements, be these under tri-therapy (Triomine: stavudine, lamivudine, névirapine) or left unattended.

Uleine; HIV; Tri-therapy; Ribonucleic Acid Oligomers (RNO)


Introduction

The treatment of AIDS patients rests on the administration of three chemicals designed to reduce the viral load. The HIV is immunodepressive but no effort is made during chemical treatment to stimulate the multiplication and activity of lymphocytes. This stimulation would however benefit the patients because some drugs are known to be immunosuppressive, which add to the immunodepression induced by the pathogen. Means exist to stimulate the immune response, either by inducing the synthesis of nitric oxide by lymphocytes and other cells, or else by promoting the multiplication of lymphocytes.

Uleine (1), present in Himatanthus lancifolius and Aspidosperma subincanum Mart. ex A. DC., Apocynaceae, have been extensively studied (Seidl et al., 2010Seidl, C., Correia, B.L., Stinghen, A.E.M., Santos, C.A.M., 2010. Acetylcholinesterase inhibitory activity of uleine from Himatanthus lancifolius. Z. Naturforsch C 65, 440-444.). Its capacity to promote the synthesis of nitric oxide (Souza et al., 2007Souza, W.M., Brehmer, F., Nakao, L.S., Stinghen, A.E.M., Santos, C.A.M., 2007. Ação da uleína sobre a produção de óxido nítrico em células RAEC e B16F10. Rev. Bras. Farmacogn. 17,191-196.) makes of these plants candidates for use as an adjuvant in the treatment of patients suffering from an immune-depression. The bark of Aspidosperma subincanum has been purified and the food supplement rich in uleine so obtained has been demonstrated innocuous (Federlin et al., 2014Federlin, J.D., Maes, D., Maes, R., 2014. Aspidosperma subincanum I. Characterisation, extraction of an uleine-enriched fraction and potential health hazard due to the contaminant ellipticine. Rev. Bras. Farmacogn. 24, 293-297.).

Nucleic acid oligomers have been shown to inhibit in vivo Shope fibroma and vaccinia viruses (Beljanski et al., 1975Beljanski, M., Chaumont, L., Bonissol, C., 1975. RNA fragments, in vivo inhibitors of Shope fibroma and vaccinia virus multiplication. C. R. Acad. Sci. Hebd. Seances Acad. Sci. D. 280, 783-789.). This effect was traced to a stimulation of the multiplication of lymphocytes and platelets (Beljanski and Plawecki, 1979Beljanski, M., Plawecki, M., 1979. Particular RNA fragments as promoters of leukocyte and platelet formation in rabbits. Exp. Cell Biol. 47, 218-225.). Lymphocyte populations depleted in vivo by cyclophosphamide responded to a stimulus of ribo-oligo-nucleotides (Beljanski et al., 1983Beljanski, M., Plawecki, M., Bourgarel, P., Beljanski, M., 1983. Leukocyte recovery with short-chain RNA fragments in cyclophosphamide-treated rabbits. Cancer Treat. Rep. 67,611-619.). This positive effect of regeneration of lymphocytes by nucleic acids observed in vivowas verified on two malignant lymphoma cases (Donadio et al., 1991Donadio, D., Lorho, R., Causse, J.E., Nawrocki, T., Beljanski, M., 1991. RNA fragments and tolerance of cytostatic treatments in haematology: a preliminary study about two non-Hodgkin malignant lymphoma cases. Deutsch. Zschr. Onkol. 23, 33-35.).

These food supplements would appear useful to help the immune system of immuno depressed patients as tuberculosis and Aids patients to counter the immunodepressive activity of the pathogens as well as the immunodepressive activity of some drugs used to eliminate the pathogen. This hypothesis was verified by administering the supplements during a year to AIDS patients. In addition, a third food supplement consisting of propolis, royal jelly and pollen, known to stimulate the immune system, was also offered to the clinician, enabling her to make a complete study of available immuno stimulators.

The clinician focused on the improvement of the health of her patients by verifying the activity of a combination of food supplements given to patients either under chemical treatment or else, due to lack of resources, left unattended. Lack of resources restricted the monitoring in the course of time of the effects of the treatments, to weight and lymphocyte numbers.

The detailed raw results (66 pages) are available (Bouillon and Ndayikunda, 2006Bouillon, C., Ndayikunda, C., 2006. Traitement complémentaire du sida par un apport naturel et traditionnel. Chalets du bonheur a.s.b. l - Belgique, 56 p (Document is available by the corresponding author.). The authors analysed and exploited them.

Materials and Methods

Uleine source and the immunostimulant

Uleine (1) was extracted from the plant Aspidosperma subincanum Mart. ex A. DC., Apocynaceae, and presented as the food supplement “Para-pau-aspido” by the Company Parabolic Biologicals (Beauvechain, Belgium) under two different concentrations: PP20 (20 mg total alkaloids) and PP 80 (80 mg total alkaloids). A supply of the products sufficient to cover a year of treatment was donated. The food supplement “Para Immuno” elaborated by the Company Parabolic biologicals was made of propolis extracted from bee-hives, Royal Jelly and pollen. This product was donated in amounts sufficient to cover a year of treatment.


2Leid is prepared by the laboratory “Laboratoire Labo-Life”. This immunostimulant to be taken sub-lingually is made of cytokines, ribonucleic acids, deoxyribonucleic acids and TGFbeta, and the product was donated in amounts sufficient to cover a year of treatment.

Triomine 530: stavudine 30 mg, lamivudine 150 mg, névirapine 20 mg, for patients whose weights was inferior to 60 kg.

Triomine 540: stavudine 40 mg, lamivudine 150 mg, névirapine 200 mg, for weights over 60 kg.

Clinical studies

Number of subjects treated was eighty, of whom sixty were included in this study. They were orphans at the orphanage “Maison Shalom”, located in the Burundi.

The files of the patients were a year old when the study was initiated in 2006 and many showed an unfavourable clinical symptomatology and opportunist infections, i.e. chronic diarrhoea, persistent fever, generalised lymphadenopathies, diverse pneumopathies, mycoses, generalised dermatoses, zona, alteration of the general state, loss of weight and asthenia. Nearly all patients had conserved their symptomatology when the treatment with supplements started and some had worsened and developed additional opportunist infections.

Among the subjects under tri-therapy since six months, some showed no clinical change, others had developed new opportunist illnesses and others were intolerant to the tri-therapy. Those patients under tri-therapy who showed no improvement and who had developed opportunistic infections that were not observed before the chemical treatment, were included in the study.

CD4’ were monitored at time one year before treatment (T-1) and at the initiation of the treatment (T0) but this monitoring could not be pursued. Weight and total lymphocyte numbers were monitored one year before treatment (T-1), at time 0 (T0), 3 months (T3) and 12 months (T12) after initiation of food supplement treatment. Treatment was stopped at T12.

The food supplement was approved by the Belgian Ministry of Health on June 23 2009 and does not require the approval of an Ethics committee for its use.

Results

Table 1 presents results of fourteen patients who were under conventional tri-therapy for about six months before addition of the food supplements to the treatment. Their state of illness was advanced. The regimen was given according to the medical imperatives defined by the treating physician. Three very ill patients under tri-therapy who died either at the beginning of the treatment or within the first three months of treatment were omitted because treatment with food supplements is a long-term process that could not have been effective at their stage of the illness. Table 2 shows the results of 46 AIDS people who were without specific medical treatment. Their state of illness was less severe than the patients under tri-therapy. The results obtained by patients under tri-therapy are presented at some length because these results are most interesting.

Table 1
Fourteen AIDS patients under conventional tri-therapy. These patients were advanced in disease, with only #9 and #10 in a conserved state. For this reason, they were chemically-treated but the treatment sometimes worsened their condition, shown by a loss in weight (e.g. #2, 11, and 14, or else a status quo: e.g. #5, #6, #10). Some underwent a severe loss of CD4 during the treatment: e.g. #6, #7, #11, #12). All gained in lymphocytes numbers during food supplement treatment.
Table 2
46 patients receiving no standard chemical therapy. Some patients were with opportunistic diseases but a sizable number were in good state or "conserved". All of them saw the number of their lymphocytes increase during treatment.

Patients under tri-therapy

P.P 80, 2Leid and P.I. and tri-therapy

The first four patients (Nbs. 1 to 4) were in a critical physical condition at time zero (T0) and received the combination [P.P 80 + 2Leid + P.I. + tri-therapy]. Patient 1 was a 19-year old girl. She weighed 30 kg at the beginning of the treatment (T0) and the number of CD4 cells had collapsed down to 6. Evidently, she did not benefit from the tri-therapy and seemed doomed. In three months of food supplements intake (T3), she gained 15 kg and her number of total lymphocytes increased from 212 to 2915. She was in good health at T12, after 12 months of the additional alternative treatment.

The tri-therapy followed from time T-1 to T0 appeared more beneficial to patients 2 to 4 because they maintained or even slightly increased in weight during that time and had retained their CD4 and lymphocyte levels, albeit in very low numbers. The food supplements [P.P 80 + 2Leid + P.I. and tri-therapy] caused a dramatic gain in weight. More importantly, there was a spectacular increase in the number of their total lymphocytes. All were in a satisfactory state of health after 12 months of food supplements intake.

P.P.80, 2Leid and tri-therapy

Patients 5 to 8 received only P.P.80 + 2Leid, in addition to the chemicals. They were in a very poor state of health before starting food supplementation: severe drops in CD4 counts, except in patient 8, low weight and low lymphocyte count. The tri-therapy had proven ineffective. After three months of food supplement intake [P.P.80 + 2Leid + tri-therapy], lymphocyte counts increased dramatically and 3 out of 4 patients had a significant gain in weight. Patient 6 also gained weight after 12 months of alternative therapy and all were in a satisfactory state of health at that moment (T12).

PP 80 and tri-therapy

Patients 9 to 12 received P.P.80 in addition to the tri-therapy. Patients 9 and 10 were clinically speaking in a “stable state”, with relatively high CD4 and lymphocyte counts at time T0. The weight of patient 9 was not measured before the start of the study, but patient 10 had a stable weight. By contrast, patients 11 and 12 had low CD4 counts despite tri-therapy and had lost weight. The tri-therapy had proven inefficient. Three months after they started to take the P.P.80 food supplement, their weight and their lymphocyte counts had increased. After twelve months of treatment with the food supplement, their health was good.

PP20, Para Immuno and tri-therapy

This regimen was given to patient 13, a 15-years old girl weighing 24 kg at the time of her admission in the orphans’ home, under chemo-therapy but with diarrhea, fever and pneumopathy and having lost ¾ of her CD4's during chemical treatment, recovered when P.P.20 and P.I. was given during three months. The use of P.P.20 instead of P.P.80, a 4-fold lower dose in active principles, resulted in a similar weight gain and lymphocyte count recovery as with the previous 12 patients, indicating that this lower dosage was just as efficacious, at least in a teenager.

Patient 14, under tri-therapy and receiving only 2Leid as food supplement had a moderate CD4 cell count but was losing weight before joining the study. Three months later, she had not only recovered but gained weight and her lymphocyte count had doubled. These results are comparable to those observed in the other 13 patients.

Patients without tri-therapy

A larger number of patients (Table 2: Nb 15 to 60) received no tri-therapy. The clinician defined their health status at time zero by qualitative criteria (conserved, good, bad etc.) and took note of opportunist infections. The loss of weight in these AIDS patients from the moment of their admission in the orphans’ home (T-1) to time zero (T0), i.e. a year later when food supplementation started, is an objective sign of their evolution, as is their viral load and the level of their CD4’. Some of them, especially young children, gained weight and increased the number of CD4 while others saw a decline in their CD4 between T-1 and T0, but only two of them (Nbs 29 and 41) had CD4 levels below the 100 mark at time T0.

Table 2 shows that, irrespective of the severity of their illness and the worsening of their physical condition during the year that preceded supplement treatment, all recovered and gained weight in addition to increasing their total lymphocyte count. All were in satisfactory or good health at the end of the twelve-month course of food-treatment. If the effects of this treatment are visible after three months in most of the treated AIDS people, some however needed more than three months of food supplement intake before showing clear evidence of recovering health (Nbs 21, 27, 56) even if an increase in numbers of total lymphocytes took already place at time T3.

Discussion

During the whole year of treatment with natural products, we deplored no collateral secondary effects except two patients on PP80, who signalled a nose blooding that disappeared after the end of the treatment. This evidence of innocuity corroborates the results obtained in animal studies (Federlin et al., 2014Federlin, J.D., Maes, D., Maes, R., 2014. Aspidosperma subincanum I. Characterisation, extraction of an uleine-enriched fraction and potential health hazard due to the contaminant ellipticine. Rev. Bras. Farmacogn. 24, 293-297.).

All applied protocols proved efficacious. The uleine boosts the synthesis of nitric oxide, the ribonucleotides stimulate the multiplication of lymphocytes and the Para Immuno (propolis, Royal jelly and pollen) stimulates the immune system. A combination of all three works best in as much that every facet of the global treatment was exploited. However, the single application of uleine to non-treated patients, be it at its highest or lowest concentration, yielded remarkable results by itself. Patients 26, 27 and 28, who were given PP80, showed a substantial improvement of their condition, at a par with that observed when complete regimens were given. This observation is valid also for patients 47 to 51, who received PP20: their condition improved at the same rate as patients handled with the two additional food supplements. The chemically unattended patients were generally in a better condition (i.e. said “conserved”) than those who were under treatment. In this group, patients 9, 10, 11 and 12 gained no weight during chemical treatment or else lost weight. Yet, PP80 given alone to them in addition to tri-therapy improved considerably their health. An explanation for this remarkable effect may be found in the fact that uleine also interferes with the inflammatory response (Nardin et al., 2008Nardin, J.M., de Souza, W.M., Lopes, J.F., Florão, A., de Moraes Santos, C.A., Weffort-Santos, A.M., 2008. Effects of Himatanthus lancifolius on human leukocyte chemotaxis and their adhesion to integrins. Planta Med. 74,1253-1258.) and regulates the immune system (Nardin et al., 2010Nardin, J.M., Lima, M.P., Machado, G.C., Hilst, L.F., Santos, C.A.M., Weffort-Santos, A.M., 2010. The uleine-rich fraction of Himathantus lancifolius blocks proliferative responses of human lymphoid cells. Planta Med. 76,697-700.). The substance possesses thus properties shared by the two other regimen proposed. One must note here that a reduction in the number of pills taken, and in the frequency of their administration, must be favoured in all cases not only for reasons of cost and burden but because it plays a considerable positive role on the psychology of the patients.

The progress registered initially on the clinical level by nearly all patients receiving food supplements was confirmed by a significant increase of total lymphocytes numbers and weight increase from the 3th month on. Patients under tri-therapy progressed most obviously at the level of secondary effects and opportunistic infections. In 2006, at the end of the supplement treatment, we registered no case of diarrhoea, no case of hyperthermia, no case of dermatose or other opportunistic illnesses. We had only three deaths by pneumocytosis, meningitis and bacterial pneumopathy but these were very advanced cases at the beginning of the treatment.

The patient under tri-therapy were given food supplements when they showed no improvement and/or developed opportunistic diseases that did not exist before the treatment with the anti-retrovirals. A tri-therapy based on stavudine, lamivudine and nevirapine is a heavy treatment that often causes secondary effects in the patient and can be a factor of health degradation in many treated subjects. Our results indicate that tri-therapy worsens the health of some AIDS patients instead of improving it, while taking food supplements not only does not cause the same negative effects but can help the patients get over those produced by conventional therapy and even cause partial recovery of AIDS patients not receiving it.

Acknowledgment

Doctor Christian Bouillon and Chloe Ndayikunda provided us with the files of the patients.

References

  • Beljanski, M., Chaumont, L., Bonissol, C., 1975. RNA fragments, in vivo inhibitors of Shope fibroma and vaccinia virus multiplication. C. R. Acad. Sci. Hebd. Seances Acad. Sci. D. 280, 783-789.
  • Beljanski, M., Plawecki, M., 1979. Particular RNA fragments as promoters of leukocyte and platelet formation in rabbits. Exp. Cell Biol. 47, 218-225.
  • Beljanski, M., Plawecki, M., Bourgarel, P., Beljanski, M., 1983. Leukocyte recovery with short-chain RNA fragments in cyclophosphamide-treated rabbits. Cancer Treat. Rep. 67,611-619.
  • Bouillon, C., Ndayikunda, C., 2006. Traitement complémentaire du sida par un apport naturel et traditionnel. Chalets du bonheur a.s.b. l - Belgique, 56 p (Document is available by the corresponding author.
  • Donadio, D., Lorho, R., Causse, J.E., Nawrocki, T., Beljanski, M., 1991. RNA fragments and tolerance of cytostatic treatments in haematology: a preliminary study about two non-Hodgkin malignant lymphoma cases. Deutsch. Zschr. Onkol. 23, 33-35.
  • Federlin, J.D., Maes, D., Maes, R., 2014. Aspidosperma subincanum I. Characterisation, extraction of an uleine-enriched fraction and potential health hazard due to the contaminant ellipticine. Rev. Bras. Farmacogn. 24, 293-297.
  • Nardin, J.M., Lima, M.P., Machado, G.C., Hilst, L.F., Santos, C.A.M., Weffort-Santos, A.M., 2010. The uleine-rich fraction of Himathantus lancifolius blocks proliferative responses of human lymphoid cells. Planta Med. 76,697-700.
  • Nardin, J.M., de Souza, W.M., Lopes, J.F., Florão, A., de Moraes Santos, C.A., Weffort-Santos, A.M., 2008. Effects of Himatanthus lancifolius on human leukocyte chemotaxis and their adhesion to integrins. Planta Med. 74,1253-1258.
  • Seidl, C., Correia, B.L., Stinghen, A.E.M., Santos, C.A.M., 2010. Acetylcholinesterase inhibitory activity of uleine from Himatanthus lancifolius Z. Naturforsch C 65, 440-444.
  • Souza, W.M., Brehmer, F., Nakao, L.S., Stinghen, A.E.M., Santos, C.A.M., 2007. Ação da uleína sobre a produção de óxido nítrico em células RAEC e B16F10. Rev. Bras. Farmacogn. 17,191-196.

Publication Dates

  • Publication in this collection
    Jan-Feb 2015

History

  • Received
    24 May 2014
  • Accepted
    04 Nov 2014
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