The Biologic Meaning of the Structural Components of Defibrotide (DF) in the Setting of HIV Disease with Refractory Herpes, Candida, Cryptosporidium Diarrhea and Multi-Organ Failure Against the Background of Down-Regulation of Cellular Immunity, Autocrine Upregulation of Inflammatory Cytokines of TNF-Alpha, IL2, IL1, IL6 and Dysregulation of cAMP/cGMP Protein Kinase A/C Signalling.

Abstract

Production of DF from native mammalian DNA involves heat controlled depolimerization of polymerized nucleic acids as a function of hyperchromicity measured at wavelengths of 260–280 nm as the ratio of Optical Density (OD) in (disorderly phase)/(OD) in (orderly phase). In double helix DNA this rearrangement can be of the order of 100%. DF is defined to represent a hyperchromicity ratio around 15%. Its structural formula is static: P1-5, (dAp)12–24, (dGp) 10–20, (dTp)13–26, (dCp)10–20. ASH Abstract #4086, 2004 reported sequence identities of the fragments at MW </=560 Da (the calculated MW of alkali Na-salt of ATP), using Reversed Phase- HPLC and (MALDI-TOF) (Oral, Lewis),to represent the capability of DF for 15% molecular rearrangement to include but not limited to: dC, dA, G, dGMP, AMP, dTTP, CTP, ATP, dGTP, CMP, cGMP, dAMP. 4 aptamers were sequenced from the PCR product of the double-stranded Defibrotide precursor (Schroer). We have analyzed Aptamer #4 of the sequence 5′ggtggtggttgtggt against the gag/pol region of HIV, and found 3 homology sites. Translation of the aptamer region in gag is a peptide “PEPTA”, and a pol gene fragment translates the same DNA sequence into TRANS. There is a conserved hairpin in the gag/pol region, just before the aptamer #4 homology. Other homologies of Aptamer #4 include but are not limited to the self-replicating portion of mitochondrial DNA (G.Gorilla, H.Sapiens), Candida tropicalis, but not Herpes, etc. T he reproducibility of these sequences in different batches of DF are still under study, and to be reported at the time of the ASH meeting. DF's mechanism of efficacy as a dose dependent modulator of 2nd messengers, however, are based on a more universally reported data. Highlighted in the literature are the HIV induced loss of bi-directional phosphorilation between Gs GTP/Gs GDP in favor of unidirectional continuous activation of adenylate cyclase (AC) by Gs GTP and secondary uninhibited upregulation of cAMP production. This leads to a known intracellular imbalance between cAMP/cGMP favoring viral replication. Also known are Beta Adrenoreceptor coupling to AC by Gs GTP, and secondary upregulation of autocrine production of inflammatory cytokines of TNF-Alpha, IL6, IL2, IL1, as well as micelle and spore formation of Candida Albicans (Sabie). Similarly, HSV-1 Alpha0 Promoter isknown to contain a putative cAMP responsive element (CRE) via activating transcriptor protein ATP/cAMP CRE binding protein CREB, NF-kB via CA ((Danaher). Reported are upregulation of cAMP, ATP, NADH, gluthathione, 2–3 DPG, NADP /NADPH, and AC by DF, albeit as static events. We will report on an in-vivo dose system aiming at resetting the broken feedback loops of bi-directional phosphorilation reactions as a common denominator for DF's mechanism of efficacy. The patient data dose-stratified from 40mg/kg/day to 300mg/kg/day using either approach are to be detailed at the time of the upcoming ASH meetings.