Hydrocarbon double-stapling remedies the proteolytic instability of a lengthy peptide therapeutic.

The pharmacologic utility of lengthy peptides can be hindered by loss of bioactive structure and rapid proteolysis, which limits bioavailability. For example, enfuvirtide (Fuzeon, T20, DP178), a 36-amino acid peptide that inhibits human immunodeficiency virus type 1 (HIV-1) infection by effectively targeting the viral fusion apparatus, has been relegated to a salvage treatment option mostly due to poor in vivo stability and lack of oral bioavailability. To overcome the proteolytic shortcomings of long peptides as therapeutics, we examined the biophysical, biological, and pharmacologic impact of inserting all-hydrocarbon staples into an HIV-1 fusion inhibitor. We find that peptide double-stapling confers striking protease resistance that translates into markedly improved pharmacokinetic properties, including oral absorption. We determined that the hydrocarbon staples create a proteolytic shield by combining reinforcement of overall alpha-helical structure, which slows the kinetics of proteolysis, with complete blockade of peptide cleavage at constrained sites in the immediate vicinity of the staple. Importantly, double-stapling also optimizes the antiviral activity of HIV-1 fusion peptides and the antiproteolytic feature extends to other therapeutic peptide templates, such as the diabetes drug exenatide (Byetta). Thus, hydrocarbon double-stapling may unlock the therapeutic potential of natural bioactive polypeptides by transforming them into structurally fortified agents with enhanced bioavailability.

Antiinflammatory effect of Oldenlandia diffusa and its constituent, hentriacontane, through suppression of caspase-1 activation in mouse peritoneal macrophages.

Oldenlandia diffusa (OD) has been used as a natural drug for the treatment of cancer in Asia and specifically in Korea. However, the antiinflammatory mechanisms employed by OD have yet to be completely understood. This study attempted to determine the effects of OD and hentriacontane, one of the constituent compounds of OD, on lipopolysaccharide (LPS)-induced inflammatory responses in mouse peritoneal macrophages. The findings of this study showed that OD inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and prostaglandin E(2) (PGE(2)). The OD inhibited the enhanced levels of cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) induced by LPS. It was shown that the antiinflammatory effect of OD occurs via the regulation of the activation of nuclear factor (NF)-κB and caspase-1. Moreover, hentriacontane was shown to ameliorate the expression of inflammatory mediators (TNF-α, IL-6, PGE(2), COX-2 and iNOS) and the activation of NF-κB and caspase-1 in LPS-stimulated peritoneal macrophages. These results provide novel insights into the pharmacological actions of OD as a potential candidate for the development of new drugs for the treatment of inflammatory diseases.

[Development of a new hydrocarbon extract from the medicinal raw material of Circassian walnut (Juglans regia) and study of its antiparasitic activity].

The authors developed a technology for preparing a hydrocarbon extract from the medicinal raw material of Circassian walnut (Juglans regia), including its green fruits, green leaves, and fresh roots. To prepare the preparation, they obtained for the first time a new extragent called petroleum Russia that was found to contain more than hundred chemical compounds by chromatography mass spectrometry. The new agent was named irillen. Experiments on albino mice and albino rats established that the new agent was low toxic. The lethal doses of irillen were calculated: LD50 was 16377 +/- 457.5 mg/kg; LD16 = 12986.4 mg/kg; LD84 was 18976.6 mg/kg for albino mice; LD50 was 16998.0 +/- 535.4 mg/kg; LD16 = 12875.3 mg/ kg; LD84 = 18583.4 mg/kg for albino rats. The irillen prepared by the authors should be referred to as a low toxic and practically nontoxic agent (Toxicity Class IV and V). Irillen has a broad spectrum of antiparasitic activity. It is effective in treating toxocariasis in dogs, larval alveolar echinococcosis, ascaridiasis, and eimeriasis in chickens, and siphachiasis.

Paraffin therapy induces a decrease in the passive stiffness of gastrocnemius muscle belly and Achilles tendon: A randomized controlled trial.

BACKGROUND: The purposes of this study were to examine the feasibility of using the MyotonPRO digital palpation device in measuring the passive stiffness of gastrocnemius muscle belly and Achilles tendon; to determine between-days test-retest reliability of MyotonPRO; and to evaluate the acute effect of paraffin therapy on stiffness measurements in healthy participants. METHODS: It is a randomized controlled trial. Twenty healthy participants (male, n = 10; female, n = 10; total, n = 20) were recruited to evaluate the passive stiffness of gastrocnemius muscle belly and Achilles tendon. Dominant and nondominant legs were randomly divided into an experimental side (20 cases) and a control side (20 cases). The experimental side received 20 minutes of paraffin therapy. RESULTS: The stiffness of muscle and tendon in the experimental side decreased significantly after paraffin therapy (P < .01), whereas no significant differences in stiffness measurements were found in the control side (P > .05). The passive stiffness of muscle and tendon was positively correlated with the ankle from 30° plantar flexion to10° dorsiflexion for dominant legs. Between-days test-retest reliability in stiffness measurements was high or very high (ICCs were above 0.737). CONCLUSION: Paraffin therapy induces a decrease in the passive stiffness of gastrocnemius muscle belly and Achilles tendon. Furthermore, the MyotonPRO can reliably determine stiffness measurements.

The Curative Role of Bitumen in Traditional Persian Medicine

AIM: Bitumen is a natural substance effusing from rocks' notches in some highland areas; it has been known as an effective remedy for treating some illnesses. Considering pain relieving properties of bitumen in traditional Persian medicine (TPM) sources, this study aims to review the viewpoints of TPM sages regarding bitumen in the context of traditional Persian medicine. It also provides applicable information for interested researchers to conduct well-designed clinical trials and evaluate therapeutic effects of bitumen claimed in TPM sources. MATERIAL AND METHOD: Various databases including Embase, SID, IRANDOC, IranMedex, Scopus and PubMed were searched with keywords "bitumen" and "Shilajit". Furthermore, main traditional Persian medicine sources including Avicenna's "Canon of medicine", "Continens Liber" by Razes, "The storehouse of medicaments" by Aghili, "Gift for the faithful" by Momen Tonekaboni and "Measure for medicine" written by Muhammad Akbar Shah Arzani were reviewed with Persian keywords "Moomiaii" and "Mumnaei" Results: According to TPM sources, bitumen was used by Iranian's physicians to treat a wide range of diseases. It was known especially as an effective remedy to improve gastrointestinal digestive problems. CONCLUSION: Bitumen is cited in traditional Persian medicine sources as an effective remedy for treatment of a wide range of diseases, especially GI disorders and bone pain. Recent studies showed the beneficial effects of bitumen in treatment of wound healing, however using it in medical practice for other health dilemma should be confirmed by conducting well-designed clinical studies in the future.

Influence of various central moieties on the hypolipidemic properties of long hydrocarbon chain diols and diacids.

A series of long (11-15) hydrocarbon chain diols and diacids with various central functional groups and terminal gem-dimethyl or -methyl/aryl substituents was synthesized and evaluated in both in vivo and in vitro assays for its potential to favorably alter lipid disorders including metabolic syndrome. Compounds were assessed for their effects on the de novo incorporation of radiolabeled acetate into lipids in primary cultures of rat hepatocytes, as well as for their effects on lipid and glycemic variables in obese female Zucker fatty rats, Crl:(ZUC)-faBR. The most active compounds were hydroxyl-substituted symmetrical diacids and diols with a 13-atom chain and terminal gem-dimethyl substituents. Furthermore, biological activity was enhanced by central substitution with O, C=O, S, S=O compared to the methylene analogues and was diminished for compounds with central functional groups such as carbamate, ester, urea, acetylmethylene, and hydroxymethylene.

[Protective creams for prophylaxis of allergic contact dermatitis]. / Kremy ochronne w profilaktyce wyprysku kontaktowego.

Occupational medicine is interested in effective prophylaxis of skin occupational diseases. Use of protective ointments and creams makes one of prophylaxis methods of professional dermatitis. The purpose of the investigation was to evaluate both the primarily irritant and the allergic action of the two creams devised by us for protection of skin exposed to aqueous solution of electrolytes. One of these creams was composed of hydrocarbons components, the other had in addition silicone oil and cetaceum. Protective properties of these creams in patients with allergic contact dermatitis were also examined. The experiments were carried out on 10 rabbits, 75 guinea pigs, and 491 patients mainly suffering from metal (Cr, Co, Ni) and formaldehyde allergy. It was found that investigated creams do not exert primarily irritant and sensitizing effects both in animals and humans. These creams demonstrate pronounced protective action in allergy due to water soluble allergens.

Inhibitors and prodrugs targeting CYP1: a novel approach in cancer prevention and therapy.

Since Human CYP1 enzymes catalyze the metabolic activation of procarcinogens and deactivation of certain anticancer drugs, the inhibition of these enzymes has been considered as an effective approach for chemoprevention and treatment of CYP1-mediated drug resistance. Recent knowledge relating to the enhanced expression of CYP1B1 in tumors also provided certain advantages in cancer therapy by the activation of prodrugs only in tumor cells. This review concentrates on the characterized CYP1 inhibitors and CYP1-activatied anticancer prodrugs. The mechanism for enzyme inhibition and activation of prodrugs, the cancer preventive/therapeutic potential of these chemicals and their related SARs are highlighted. According to their structural features, CYP1 inhibitors are divided into the following categories: flavonoids, trans-stilbenes, coumarins, terpenoids, alkaloids, quinones, isothiocyanates and synthetic aromatics. In the same way, CYP1-activatied prodrugs are categorized into four groups: benzothiazoles, flavonoids, stilbenes and alkylating agents. Almost all of these inhibitors and prodrugs are planar molecules with one aromatic ring and some have similarity with identified CYP1 substrates. CYP1 inhibitors could effectively block the procarcinogen-induced tumor initiation in animal models and benefit us with chemoprevention. The advent of Phortress and aminoflavone as clinical candidates shows promising perspectives in developing CYP1-mediated prodrugs as chemotherapeutic drugs that are specifically activated in tumors. All of these preclinical and clinical studies indicate that inhibitors and prodrugs target CYP1 are promising anticancer strategies.

Palmitone isolated from Annona diversifolia induces an anxiolytic-like effect in mice.

The aim of this study was to investigate the behavioral effects of palmitone in the anti-anxiety response in experimental models in mice. In the elevated plus-maze test, palmitone (0.3, 1, 3, 10 and 30 mg/kg, I. P.) lengthened, from 50 % to 199 %, the time spent in the open arm region of the maze at all doses tested, as compared to the vehicle group ( P < 0.001). In relation to the rearing activity in the exploratory cylinder, palmitone significantly modified ( P < 0.05), in a dose-dependent manner, this activity by decreasing the number of rearings with an effective dose value (ED (50)) and 95 % confidence limits (CL (50)) of 0.79 (0.23 - 2.68) mg/kg. In addition, in the hole-board test, nose-poking was also significantly decreased ( P < 0.01) in a dose-dependent fashion [ED (50) (CL (50)) = 9.07 (4.51 - 18.26) mg/kg]. Moreover, palmitone at any dose caused no change in motor activity nor disruption in traction performance. In contrast, diazepam, used as reference drug, produced an anxiolytic effect with a significant and dose-dependent decrease in motor coordination accompanied by disruption of the traction performance. Behavioral studies suggest an anti-anxiety effect produced by palmitone, but its neuropharmacological profile differs from that observed for benzodiazepines such as diazepam.

Anticonvulsant effect of Annona diversifolia Saff. and palmitone on penicillin-induced convulsive activity. A behavioral and EEG study in rats.

PURPOSE: To evaluate hypnotic and anticonvulsant activities of Annona diversifolia Saff. and palmitone by using behavior and electroencephalographic (EEG) analysis in an experimental model of focal seizures in rats. METHODS: For hypnotic assessment, EEG analysis of polysomnographic slow-wave sleep (SWS) and rapid eye movement (REM) sleep for a 1 h period were performed after vehicle, A. diversifolia extract or palmitone, administration. For anticonvulsant effect, 60 minutes after treatments, EEG and behavior were analyzed during penicillin-induced seizures. Latency to the onset of the first paroxystic spike, first seizure and frequency, as well as seizure severity using Racine's scale, were determined. RESULTS: Palmitone, but not A. diversifolia extract, produced a delay in the latency to the SWS phase. In addition, both palmitone and extract decreased SWS duration and accumulated REM sleep phase. With regard to the seizures, both the extract and palmitone increased the latency to the onset of spikes and seizures, but also decreased the duration of penicillin-induced seizures. This reduction in the EEG recordings was associated with an attenuation in the severity of behavioral seizures. CONCLUSIONS: A. diversifolia and palmitone did not produce a sedative-hypnotic effect although both of them were effective in reducing the severity of behavioral and EEG seizures induced by penicillin in rats, suggesting that the diminution in the paroxystic activity by A. diversifolia is likely produced by palmitone through GABAergic neurotransmission. This study justifies and reinforces the traditional use of this plant in epilepsy.

M2000: a revolution in pharmacology.

BACKGROUND: The tolerability and the anti-inflammatory and immunosuppressive properties of a novel designed non-steroidal anti-inflammatory drug, M2000 (beta-D-mannuronic acid), were investigated in various experimental models. MATERIAL/METHODS: The anti-inflammatory and immunosuppressive properties of M2000 were tested in experimental models of rheumatoid arthritis (AIA) and multiple sclerosis (EAE). Its therapeutic potency on kidney diseases was studied using experimental models of nephrosis and immune complex glomerulonephritis (ICG). Biocompatibility and pharmacotoxicology assessment of M2000 was carried out using a fibrosarcoma cell line, zymography, and serum and urine determinants. RESULTS: Data showed that oral and/or i.p. administration of M2000 significantly reduces paw edema and histopathological parameters in arthritic rats. The immunosuppressive property of M2000 could significantly diminish clinical signs and histological erosions in the EAE model. Lymph node cell proliferation assay in EAE confirmed the immunosuppressive efficacy of the tested drug. Our findings in ICG and experimental nephrosis showed that M2000 enables a significant decrease in proteinuria, BUN, serum creatinine and cholesterol, as well as glomerular lesion in M2000-treated rats. Moreover, this drug inhibited MMP-2 activity. The pharmacotoxicology study showed that M2000 is the safest anti-inflammatory and immunosuppressive drug in comparison with dexamethasone and conventional NSAIDs tested. Additionally, M2000 had no ulcerogenic effect on the rat stomach. CONCLUSIONS: M2000 is the first novel designed NSAID with the lowest molecular weight, no gastro-nephrotoxicity, and therapeutic effects in glomerulonephritis and nephrosis and could be strongly recommended on an extensive scale as the safest drug for decreasing anti-inflammatory reactions.

Larvicidal effects of mineral turpentine, low aromatic white spirits, aqueous extracts of Cassia alata, and aqueous extracts, ethanolic extracts and essential oil of betel leaf (Piper betle) on Chrysomya megacephala.

BACKGROUND: Many methods have been employed, with variable success, in the treatment of cutaneous myiasis caused by Chrysomya species. AIMS: Experiment 1: to assess the larvicidal effect of mineral turpentine (MT) and the main ingredient of MT, low aromatic white spirits (LAWS), on Chrysomya megacephala larvae in vitro. Experiment 2: to assess the larvicidal effects of aqueous extracts of winged senna (Cassia alata), and aqueous extracts, ethanolic extracts and essential oil of betel leaf (Piper betle). METHODS: In experiment 1, two samples of LAWS were obtained from two industrialists (samples 1 and 2). Adult flies of C. megacephala were bred in the insectory of the Department of Parasitology, Faculty of Medicine, University of Colombo. Petri dishes were prepared with pads of cotton wool. These cotton pads were soaked separately in MT, LAWS samples 1 and 2, and normal saline as a control. Ten larvae were placed in each Petri dish. The activity of the larvae was observed and recorded half-hourly. MT and the two samples of LAWS were analyzed by chromatography. In experiment 2, volatile essential oil of betel was prepared using a standard steam distillation process. An ethanolic extract of betel was obtained after boiling the crushed leaf with water, and mixing the stock with ethanol. Betel oil dilutions of 1-4% were prepared using 1% Tween 80 (v/v aq) as a solvent, with 0.05 g/100 mL sodium lauryl sulphate (as stabilizer) and 0.01 g/100 mL methyl paraben (as a preservative). Cotton wool swabs soaked in 1, 2, 3 and 4% essential oil of betel in 1% Tween 80 (v/v aq) prepared as above, 1, 2, 3 and 4% ethanolic extract of betel, 50 and 25% aqueous extract of C. alata, and 50 and 25% aqueous extract of betel were placed in separate Petri dishes. Ten larvae were placed in each Petri dish. 1% Tween 80 solvent with the stabilizer and the preservative, but without betel essential oil, was used as a negative control and MT was used as a positive control. Larval motility was assessed as before. RESULTS: MT and the two LAWS samples killed the larvae in vitro within 4 h. Chromatography showed more unidentified constituents in MT than in pure LAWS, indicating additional substances in MT. The 4 and 3% preparations of the essential oil of betel were effective in killing 100% of the larvae of Chrysomya within 3 h 30 min. The 2% extract of betel essential oil killed 96.7% of larvae in 4 h. Ethanolic and aqueous extracts of betel, the aqueous extract of C. alata, normal saline and the Tween 80 solvent were not larvicidal. CONCLUSIONS: MT and LAWS, the main ingredient of MT, were effective in killing Chrysomya larvae. Essential oil obtained from betel leaves also showed a dose-dependent larvicidal effect on Chrysomya larvae. This natural product may be effective in the treatment of wound myiasis.

M-2000, as a new anti-inflammatory molecule in treatment of experimental nephrosis.

The therapeutic effect of M-2000 (C6H10O7) molecule was tested in Adriamycin-induced nephropathy. To induce experimental nephrosis, Adriamycin was given once by a single intravenous injection (7.5 mg/kg) through the tail vein. Six days after injection of Adriamycin, therapeutic protocol was developed by intraperitoneally (i.p) administration of 30 mg/kg M-2000 solution. Total of i.p. injections were 14, in which five injections were made every day and nine injections were carried out at regular 48-h intervals. Therapeutic protocol was terminated on day 28 and animals were killed on day 43. The treated patient rats showed a significant reduction in proteinuria, BUN, serum creatinine and serum cholesterol, as well as, administration of M-2000 could significantly diminish the serum level of interleukin-6 (IL-6) in treated animals compared to non-treated controls. Moreover, treatment with M-2000 significantly reduced number of glomerular leukocytes, Hypercellularity and hydropic change in capillary network within the renal cortex and decreased tubular casts. These data suggest that M-2000 therapy can ameliorate proteinuria, and suppress the progression of glomerular lesions in experimental model of nephrosis.