PatentView-2-2016

Expiring Patents	Notes
CA 2 223 616 Hypersensitive Response Induced Resistance in Plants FD 05-06-1996 PD 07-06-1995 ED 05-06-2016 Cornell Research Foundation Inc., USA	Cell death and tissue necrosis at sites of microbial infection (e.g. rust disease) prevent propagation of disease, reproduction of pathogenic microbes (viruses, bacteria, fungi) in plants. The result is resistance induced in plants by hypersensitive response (HR). The process is an interaction among products of plant resistance genes and that of microbial avirulent genes. The plant HR is species-specific both for plants and microbes in the interaction. Essence of the invention: HR induced microbial resistance in monocotyledons and dicotyledons (grain crops and ornamentals as well). Microbial source of polypeptides/proteins eliciting HR in plants: Erwinia amylovora, Erwinia chrysanthemi, Pseudomonas syringae, Pseudomonas solancearum, Xanthomonas campestris. The process: transformation of nonpathogenic bacterial expression system (e.g. E.coli) with gene sequences coding for polypeptides/proteins eliciting HR > contact application of the bacterial expression system, in plants. preparation of recombinant polypeptide/protein eliciting HR > contact application (spray, injection, scrape) in liquid vehicle (water) supplemented with accessory components (fungicides, insecticides...). Bio-perspective: in the EU regulation issued in January, 2016 the maximum levels of pesticide residues in/on food of plant and animal origin, are definitely restricted. Consequently, alternatives like HR induced resistance in crop protection are given a higher emphasis. Parameters to be clarified are: HR induced resistance depends on host plant genetic assembly... HR induced resistance depends on environmental factors, on breeding in open field circumstances... time period of HR induced resistance? Acceptance of new induction on an existing one? competitiveness of HR induced resistance with efficiency of pesticides?
GB 2 301 775 High Dosage Lutein and Zeaxanthin for Macula Therapy FD 07-06-1996 PD 07-06-1995 ED 06-06-2016 The Howard Foundation, GB	Approximately 20% of population aged over 65 are touched by visual impairment, the so-called age related macula degeneration (AMD). Regarding its anatomical situation, the macula is part of the retina and contains pigments (> xantophylls, > macular pigments) protecting tissues beneath. Pigments of the macula have an absorption spectrum characteristic to the carotenoids, suggesting the presence of lutein and zeaxanthin (isomers) in varying ratio. As for the gradient: predominance of zeaxanthin in macula area alters into predominance of lutein in the retinal periphery. According to confirmatory measurements, lutein/zeaxanthin content of AMD retina showed a level 30% lower than that of normal retina control. The purpose in therapy and prophylaxis is the adequate supplementation of retinal piments. Essence of the invention: target specific pharmacological formulation of antioxidant lutein/zeaxanthin (in ester) for effective absorption in combination with additional synergistic vitamins and mineral components. Source for lutein/zeaxanthin production are edible (green, coloured) plants containing carotenoids. Production is regulated/increased by genetically modified plants (e.g. GM tomato) targeted with expression vectors (plasmids) containing DNA sequences coding for key enzymes (four enzymes) in the biosynthesis of carotenoids Bio-perspective: experimental findings in vivo and in vitro may lead to broadened area and mode of application: diabetes retinopathy, cataract, uveal melanoma, phototoxicity, retinal ditachment, uveitis... delivery per os, in eye droplets, by intravitreal injection... turn to higher doses.
CA 1 340 580 Recombinant Alveolar Surfactant Protein FD 11-12-1985 PD 11-12-1984 ED 08-06-2016 BYK Gulden Lomberg Chemische Fabrik GmbH, Germany	Free blood-air transport on alveolar surfaces in the lung is assured by the formation and deposition of surfactants, i.e. protein complexes preventing alveolar collapse in exhalation. Constituents of the complex are the alveolar surfactant protein (ASP) synthesized by type II alveolar cells, and phospholipids. Failures in formation of the surfactant complex (infant and adult respiratory distress syndrome > RDS > hyaline membrane disease > idiopathic RDS, adult bronchopulmonary dysplasia) is an implication for replacement therapy. ASP is a conserved protein along mammalian evolution. BUT: isolates of animal origin are unsteady in composition + there are infectous hazards not to pass by. Ultimately, standard and controlled preparation of recombinant ASP was realized, and the product underwent combination with phospholipids in reaching the final form. Method: insertion of cDNAs (DNA II-IV exons + portion of DNA I exon) coding for ASP (32kD apoprotein) and for a hydrophobic protein (10-20 kD) into expression vector, the latter delivered into producer host cell. The produced protein was harvested, purified, and entered (Ca2+) into complex with synthetic phospholipids. Then, pharmacological formulation of the complex followed. Bio-perspective: resolving drawbacks of the recombinant products compared to natural surfactants; active substance: finding optimum ratio of apoproteins and hydrophobic proteins, optimum ratio of proteins and phospholipids (counteracting phospholipase activity) . delivery/formulation: tracheal instillation > endotracheal tube related complications, aerosol of particle size 0,5-2,0 µm > aerosol only enters ventilated alveoli. RDS and ALI (acute pulmonary injury) represent distinct solutions in therapy. need for personalized formulation of the complex. need for personalized delivery of the complex.
CA 2 224 565 Agent for Combating Parasites in Farmed Fish FD 10-06-1996 PD 13-06-1995 ED 10-06-2016 Nutreco Aquaculture Research Centre A/S, Norway	From public health and economic point of view, maintaining fish stocks in healthy conditions is among the basic requirements in fish farming. The same applies for sea salmon farms (e.g. Salmon salar), too. In the latter, priority is given to making fish farms free of sea lice Lepeophtheirus salmonis (Arthropoda > Crustacea, family Caligidae), parasites in juvenile phase of development, feeding on fish body surface mucus, skin and blood. Infestations due to these parasites manifest by direct and indirect ways, as well. Directly, these parasites make wounds on skin of salmon (aquafarm of 6^oC - 7^oC ), giving way to infections through them. Indirectly, microbial pathogens (mainly viruses, worms) themselves, carried by these parasite vectors and discharged in their secretions, infect the target fish or, together with the parasite vector, they enter the target fish organism at wounds or, through the mouth and the gills. Previous approaches in protective use of insecticides (e.g. formaline, hydrogen peroxide, macrolid antibiotics, organophosphates dissolved in water of aquafarms, synthetic pyrethroids in oil-emulsion formulations) proved to be insufficient in aspects of toxicity, labour demand, and of developing resistance. Essence of the invention: teflubenzuron embedded in or coating feeding pellets/granules /powder for the inhibition of arthropodal chitin synthesis > disturbing UDP-N-acetyl-glucosamine transport in epidermal cells in any phase of development > restrain of moulting, leading to situation incompatible with life of crustaceans. Bio-perspective:teflubenzuron is not the ideal solution. N-[(3,5-dichloro-2,4-difluorophenyl)carbamoyl]-2,6-difluorobenzamide (C₁₄H₆Cl₂F₄N₂O₂)_:is hardly soluble in water > associates to organic substances > low bioavailability in fish, hardly metabolized > excreted in original form > contribution to sediment formation > long life of drug in seabed (half life cca 115 days) > leaching / passive transport to nearby areas. Issue of target specificity: developmental abnormalities arisen in wild populations with chitin-exoskeleton living in proximity of coastal fish aquafarms. Driven by environmental factors back-and-forth transfer of parasites between populations of coastal fish aquafarm and of wild habitat, leading to higher mortality in populations of wild habitat. UK-Chile: inventive program on maintaining Atlantic salmon lice-free. Priorities: improving host organisms' resistance to parasites; finding optimum in nutrition to reach resistance required.
US 5,912,013 EQUETRO carbamazepine capsule XR CARBATROL carbamazepine capsule XR FD 21-04-1995 PD 23-07-1991 PD 23-07-1992 ED 15-06-2016 Validus Pharm. LLC & Shire US Inc.	Active substance Carbamazepine is a tricyclic substance with antidepressive, antiepileptic (> epileptic grand mal), analgetic (trigeminal neuralgia, diabetes neuropathia) and antidiuretic (indirectly on hypothalamus or directly on renal tubules?) effects. The mechanism of action is unclear. The interference of carbamazepine with action potentials by inactivation, inhibition of Na+ channels in presynaptic and postsynaptic membranes of CNS synapses, is presumed. At biochemistry level: carbamazepine acts as an inducer of cytochrome CYP450 3A4/1A2/2B6/2C9/2C19 enzymes. It down regulates transcriptional activity of cPLA2 (arachidonic acid selective cytosolic phospholipase A2) gene by down regulating the activity (DNA binding) of transcriptional factor AP-2 (the latter is achieved by cAMP dependent PKA pathway). Essence of the technical invention Maintaining blood level of carbamazepine (active substances) in therapeutic range. Formulation of one multiple-unit dose comprising three units with different substance release characteristics. The first of them is an immediate release (IR) unit. The second is a sustained release (SR) unit withstanding pH changes in the gastrointestinal tract and, providing sequential release of substance over 6-10 h residence time. Similar to the second, the third is also a sustained-extended release unit (XR), sensitive to pH of the lower gastrointestinal tract when releasing substance. Details of the inner and surface formulations of the units and related professional arguments are disclosed. Bio-perspective: extended release remains a demand (see per os mono- and combination therapy) prevention: daily dose in one delivery, in even dosage chronic diseases: maintaining blood level of active substances in therapeutic range nanotech hydrogel vehicles
EP 0 854 843 Controlling Wastewater Treatment By Monitoring Oxygen Utilisation Rates FD 21-06-1996 PD 22-06-1995 ED 20-06-2016 SFC Environmental Technologies Pvt. Ltd.	Recovery of wastewater by regulated biological/microbial removal of N/C/P from sludge biomass; a method widespread in our days. Essential part of the system is the apparatus comprising an open, two-zone reactor (aerobic fermenter) with zones related to each other. The first zone is a receptacle for incoming waste, the second is the receptacle for the mass coming from the first zone (partial recycling to the first zone has functional advantage). It is the second zone of the reactor where microbial (nitrificating, denitrificating bacteria...) metabolic processes are regulated by cyclic aeration and introduction of dissolved oxygen into the biomass. Electronic oxygen sensors for monitoring O₂consumption are located in the second zone or in the pipe for discharge of the mass. Bio-perspective: flocculating microbial components decreasing efficiency of the method, are changed for biofilm formation with microbes dedicated for metabolic processes in demand; target specific (> size, > material) carriers for biofilms Zero Liquid Discharge - ZLD: recovery and reuse of wastewater in increasing demand for fresh water: is it the future?
US 5,260,440 ¬¬ US RE 37314 CRESTOR rosuvastatin calcium oral tablet FD 12-06-1992 RE 27-08-1998 PD 01-07-1990 ED 08-07-2016 IPR Pharmaceuticals Inc.	Essence of the invention: description of the synthesis of pyrimidine derivative rosuvastatin-Ca {second generation -synthetic- statin C₄₄H₅₄CaF₂N₆O₁₂S₂}. Function: statins are selective and competitive inhibitors of HMG-CoA reductase (3-hydroxy-3-methyl-glutaryl-coenzymeA reductase), the enzyme catalyzing the step HMG-CoA → mevalonate in cholesterol synthesis. Further, statins enhance LDL (low density lipoprotein) receptor expression on liver cell surface. The aforementioned result in decrease of hepatic VLDL synthesis and, by entering of LDL cholesterol (LDL-C) from blood into liver cells (aiming catabolism) the level of LDL-C in circulation will be reduced [indication for use: familiar and acquired hypercholesterolemia, stroke, CVD therapy, prevention]. An example of statin efficacy in reducing LDL level in Qatar (2013): rosuvastatin (10 mg) 29,03%; atorvastatin (40 mg) 22,8%; pravastatin (20 mg) 20,3% . Bio-perspective: A/ Technology: generation of gem-difluoromethylenated rosuvastatin analogues by inserting functional groups in the hydrophilic side chain. According to preliminary information, the inhibition of enzyme activity > 50%, performed by very low doses of statins concerned (in vitro test, molecular docking evidence - Zhao Zhao et al. 2016). B/ Function: statin pleiotropy? S tabilizing atherosclerotic plaques: plaques arisen in the aortic vessel wall comprise of lipid core covered with fibrous cap, the latter mainly composed -among others- of collagens type I and III synthesized by aortic smooth muscle cells (key enzyme prolyl-4-hydroxylase/P4Hα1). Stability of the plaques depends also on the degrading activity of matrix metalloproteinases synthesized by macrophages and smooth muscle cells (MMP2, MMP9). The proinflammatory CD40L (tumor necrosis factor receptor/TNF-R family) induced repression of P4Hα1 (> repression of collagen synthesis) is attenuated by rosuvastatin inhibiting the signaling pathway TRAF6-JNK-NF-κB switched on by CD40L, in human aorta smooth muscle cell culture. Neuroprotection: rosuvastatin given per os in rat experimental glaucoma model led to a decrease in loss of retinal ganglion cell number, in apoptosis. The result was not achieved with rosuvastatin given in intravitreal dose. C/ New directions Proprotein convertase subtilisin/kexin type 9 serin protease PCSK9 is a catalyst of different proenzyme → enzyme conversions (see: hormones, growth factors, cytokines, cell surface receptors in proteolytic maturation). PCSK9 is highly expressed in liver, intestines, kidneys, and the central nervous system. Intracellular PCSK9 in liver cells promotes LDL receptor (LDLR) maturation (pro-PCSK9 chaperon + LDLR precursor in mutual catalytic maturation along the ER > Golgi > cell surface directed transport). Secreted PCSK9 is a posttranslational regulator of cell surface LDLR number. By binding to EGF-A region of LDLR, PCSK9 gets into complex with LDLR and by clathrin mediated endocytosis, this complex reaches the endosomal-lysosomal (degrading) compartment. The latter providing requirements in molecular conformation and pH, make a stop to LDLR recycling to the cell surface ultimately leading to reduction in the number of cell surface LDLR. Besides LDLR gene expression, PCSK9 gene expression too, are enhanced by statins. Nonsense mutations in PCSK9 gene result significant decrease in plasma LDL-C mean values. 426C→G encoding Y142X, a tyrosine > stop codon replacement at position 142. 2037C→A encoding C679X, a cysteine > stop codon replacement at position 679. One of the nonsense mutations above have 2% frequency in black populations; the resulting phenotype is a decrease by 40% in plasma LDL-C mean values. *137G→T encoding R46L, an arginine > leucine replacement at position 46. This latter nonsense mutation has 3,2% and 0,6% frequency in white and black populations, respectively; the resulting phenotype is a decrease by 21% in plasma LDL-C mean values. The corresponding new inventive approaches are: monoclonal antibody against PCSK9 (PCSK9-mAB) to bypass the catalytic action and to support statin therapy. OSLER clinical trials // Amgen // Evolocumab (human mAB) // Repatha™ (EMA 17-07-2015; FDA 27-08-2015) Phase III study (12-52 weeks): well tolerated statin therapy along with a diet + Evolocumab > decrease by roughly 60% in plasma LDL-C level in all groups under treatment. Homozygous familiar hypercholesterolemia: decrease by 31% in plasma LDL-C level. In process: ID. NCT01624142 and further clinical trials. ODYSSEY clinical trials // Sanofi + Regeneron Pharm. // Alirocumab (human mAB) // Praluent® (FDA 24-07-2015; EMA 23-09-2015) Placebo controlled trial (78 weeks): compared to placebo effect, Alirocumab resulted decrease by 62% in plasma LDL-C level (patients with heterozygous familiar hypercholesterolemia + other cardiovascular threats). In process: ID. NCT01604824 and further clinical trials. SPIRE clinical trials // Pfizer // Bococizumab (humanized mAB) Phase III study results are expected in 2017. In Phase II studies humanized mAB decreased plasma LDL-C level likewise to human mABs above. permanent loss-of-function mutation in PCSK9 gene by gene editing CRISPR-CAS9 system. Coexpressing {GFP (green fluorescent protein) + guideRNA (specificity to PCSK9 exon 1, 2) + Cas9 (nuclease)} adenovirus given to experimental mice result in three days in: decrease by 35-40% of plasma cholesterol level, reduction of circulating PCSK9 level, enhancement of LDLR exposure on liver cell surfaces. Related questions: safety of using adenovirus vector? / off-target mutagenesis in probability? geographical and lifestyle parameters in developing cardiovascular anomalies. ATP citrate lyase (ACL) links glucose metabolism to lipid synthesis; it catalyzes cytosolic acetyl-CoA synthesis. Apart from statin transporters, the chemical small molecule 'Bempedoic acid' (ETC-1002 // Esperion) is taken up by different mechanisms in the liver where it undergoes transformation and the derivative ETC-1002-CoA is generated. This latter by direct inhibition on ACL, impedes provision of substrates to cholesterol and fatty acid synthesis. There are already 10 safety-efficacy studies performed so far by Esperion, having involved among others patients with dyslipidemia, hypercholesterolemia, type 2 diabetes. Aftermath of the Esperion phrase 'statin intolerance' was a regulatory dialogue with FDA to define the phrase in essence. Further clinical studies are needed for assessment of the product ETC-1002 in dedicated patient populations. In process: ID. NCT02659397 clinical trial for the pharmacokinetic, pharmacodynamic, and safety analysis of placebo controlled ETC-1002 + Atorvastatin treatment.
EP 0 754 764 Method and Reagent for the Specific Determination of mRNA FD 18-07-1996 PD 21-07-1995 ED 17-07-2016 Roche Diagnostics GmbH Mannheim, DE	Essence of the invention: specific detection of 3'-poliA tailed nucleic acid sequences, i.e. eukaryotic mRNA sequences in cells and tissue samples without prior isolation or purification. According to the method, labeled oligonucleotide probe containing complementary sequence to the 3'-poliA tail or to the target nucleotide sequence, is added to the sample disrupted prior with lysis and/or hybridizing buffer (> knowledge of target sequences, at least in part, is a must). Oligonucleotide probes are labeled by biotin, and, correspondingly, the reaction vessel wall is covered by streptavidin/avidin. Target sequences captured then go through reverse transcription (> cDNA generation), thereafter DNA amplification driven by specific DNA primers and heatstable DNA polymerase (RT-PCR reaction) will follow. The end product is characterized in electrophoresis. The method/process is ready for automation. Bio-perspective: beyond detecting expression of some genes, more than 15 years ago, the microarray (microchip) technique capable for simultaneous analysis of thousands of genes/gene products of differential gene expression, became part of the genetics/genomics toolkit. Points to consider: For probes purchased, or produced in house, microarray detections also rely at least on partial knowledge of target sequences. Microarray detections require DNA in microgram quantities provided by PCR, a method not free of sample bias. Applicability of microarrays is limited by cross hybridization of similar sequences making analysis of allelic variations, SNPs (small nucleotide polymorphisms), alternative splicing variations and other alterations more difficult. For clearing up the difficulties mentioned before, new methods based on nucleic acid sequencing, the NGS, RNA-seq (next generation sequencing, RNA-sequencing) techniques dedicated for whole genome/whole transcriptome analysis were developed (and are under further development). These are techniques making it not necessary to know at least partially the target sequences, and, they require DNAs in nanogram quantities, hence, the need for PCR is minimal, if at all.
CA 2 183 108 Algae Control in Swimming Pools FD 12-08-1996 ED 12-08-2016 Sani-Marc Inc. Canada	Algae colonizing swimming pools are hosts for potential microbial pathogens; elimination of them blocks chain reactions unwished. Conventional disinfection of swimming pools deals with chlorine compounds (calcium hypochlorite, lithium hypochlorite, sodium dichloro-s-triazinetrione); derivatives of them in aqueous media {x(OCL)yz ; OCl^_} are toxic to algae too, among others. When disinfecting chlorine (or bromine) compounds are in use, the adjustment of pH (7,2> pH <7,6) is needed regularly, mostly with adding soda. However, this latter in overdose necessitates further an acidic component to reach the pH required. The consequences are → high load of irritating (to the eyes, skin, lungs, environment) chemicals in the pool water and, in the vapor above it. Essence of the invention: Zn component (e.g. ZnO) added to chlorine compounds {chlorine compound 0,5% (w/w) < Zn component < chlorine compound 50% (w/w)} enhances biocidal effect of the latter, consequently, the quantity of chlorine compounds is to be reduced for reaching optimum results (an example of weight ratio: 1 g ZnO / 200 g Ca-hypochlorite). For the elimination of residential opalescence of pool water (due to particles smaller than filter pore size) remained after physical filtration, accessory collecting component ('particle collection agent') is added to the water. This latter component (preferably aluminium salts) collects remnant particles by flocculation/precipitation/coagulation, making them ready for final mechanical removal. Bio-perspective: results of complex analysis (gas chromatography, mass spectrometry, Salmonella mutagenicity test/Ames test) carried out on water samples taken from pools, recreational spas disinfected with chlorine, bromine coumpounds, give professional arguments in favor of algae (and microbial) control free of chemicals used. after physical filters, the insertion of ultrasound alone, or in combination with H₂O_2 . after physical filters, the insertion of ultraviolet irradiation. ionization by low direct current with silver and copper alloy electrodes; passing water is enriched in Ag⁺ and Cu²⁺ metal ions inhibitory to bacterial and algal growth/reproduction (non-competitive enzyme inhibition, DNA-binding).
US 5,585,135 Method for Extending the Shelf-Life of Chocolate Confectionery Products Containing Peanuts and the Product Produced Therefrom FD 24-08-1995 ED 21-08-2016 The Hershey Company PA/USA	Quality and acceptibility of most confectionery products are significantly influenced by their fat and oil content. In case of polyphenol-rich chocolate, perceptual quality comes along with rigorous requirements even at primary commodities level {> origin and quality of cocoa bean (Theobroma cacao), of milk powder, of peanut (Arachis hypogaea) additives, impact of moisture content (low moisture beans with potential of Salmonella heat tolerance). For hazelnut chocolates, prior to admixing peanuts are processed, preferably by roasting (in air or, in oil). Advantages of roasting: food hygiene peanuts beany taste is turned off inactivation of oxidative enzymes (like lipoxygenases oxidising fatty acids' unsaturated bonds) > longer shelf life. Disadvantages of roasting: it is also detrimental to antioxidants (e.g. to the enzyme superoxide dismutase) oil compartments in the cells are destroyed, agglomerations of oil come into contact with oxidising components (e.g. Cu²⁺, Fe²⁺, Fe³⁺ ions) giving rise to fast rancidity (see: shelf life of solid milk chocolate > 1 year, of milk chocolate containing peanut < 8 months). Instability (oxidative rancidity) of peanut oil is due to the relatively high proportion of polyunsaturated fatty acid linoleic acid (18:2). 'flavor fade' effect (oxidation/rancidity products → peroxides → accumulation of aldehydes, hydrocarbons). Lipophilic items of oxidative decomposition in peanuts may act as cross-contamination too, by dissolving in the lipid phase of chocolates. To avoid this latter, in theory, there are more options to choose: antioxidants in overdose added to the product/deaeration during production/nitrogen flushing/storage under inert gas. However, neither is viable in practice. Essence of the invention: production of chocolates containing peanuts (5% - 60% w/w) with improved quality and shelf life. Peanuts of high oleic acid (18:1) content (HOAP) are produced with low share of linoleic acids (18:2). HOAP are processed in shell after blanching or without it (in whole, split, chopped). Key point in the invention is the production of peanut F1250 obtained by repeated genetic backcross resulting higher quality as of 80% oleic acid (in contrast to 40-50% of conventional nuts), with larger bean size, with closed shell. F1250 is less sensitive to oxidative rancidity, comes without 'flavor flade' but with longer shelf life than before. Bio-perspective: Peanut component Restriction of aflatoxin (Aspergillus toxin) synthesis by probiotic strains Saccharomyces and Lactobacillus. Mapping of polygenes determining major and minor fatty acid synthesis, search for decisive steps in regulation. Cocoa-chocolate component: what about us, cocoa tree? http://www.scientificamerican.com/article/the-race-to-save-chocolate/ Cocoa tree (cocoa bean) preferences: hot + shady + wet climate (within the 20. latitude), well drained soil (scarce in the tropics). Cocoa production 70%: Ivory Coast > Ghana > Nigeria > Cameroon Others: Indonesia, Brazil, Ecuador The way from cocoa bean to chocolate covers 50 million peoples's livelihood. Cocoa production: more than 4 million metric tons/year. Estimation for 2020: supply is surpassed by 1 million metric tons in demand. Fungal, viral, rots diseases, extremes in climate due to global changes: approximately 40% loss in cocoa harvest. Harvest in majority is provided by small farms contesting climate, soil quality, accession to fertilizers, irrigation, phytoparasites, expenses, sales, legal deficiencies, migration of young people, undereducation. Ways to ascend: Fair Trade Foundations World Cocoa Foundation (Bill & Melinda Gates Foundation + 17 companies) Nestlé's initiative Hershey's responsibility Quality and yield, diversity of cocoa trees depending on genotype
US 5,955,109 Methods and Compositions for Topical Delivery of Retinoic Acid RETIN-A MICRO tretinoin Gel microsphere topical FD 17-06-1993 PD 18-12-1985 ED 21-09-2016 Valeant Pharmaceuticals International Inc.	Topical gel dedicated for acne vulgaris appearing most frequently and intensely in late adolescence. The active substance is the vitamin A metabolite tretinoin (> retinoids), or {(all-E)-3,7-dimethyl-9-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2,4,6,8-nonatetraenoic acid} as of IUPAC, or all-trans-retinoic acid (C₂₀H₂₈O₂). Essence of the invention: gel of microporous structured crosslinked polymer methyl methacrylate/glycol dimethacrylate, with active substance tretinoin and supplementary components carried in the micropores. On treated surfaces the active substance is released from the pores according to law of diffusion. Besides photoprotection of tretinoin, depending on the pore size micropores support the controlled, extended release of the active substance. Bio-perspective: Acne vulgaris is a disorder related to the skin pilosebaceous unit (PSU). PSU = basal membrane boundered anatomical unit comprising hair follicle with the squamous cells covered follicular canal + muscle filament (erector pili) + sebaceous gland. Upon androgenic (profoundly DHT=dihydrotestosterone) hormone stimuli , the epithelial stem cells lining PSU basal membrane give rise to differentiating daughter cells explaining the excessive accumulation of keratinocytes, sebocytes and the overproduction of sebum, all the consequences of hormonal surges in adolescents. According to present knowledge, the proliferation and differentiation into keratinocyte and sebocyte cells are mainly due to the effects of metabolic determined IGF-1 (Insulin-Like Growth Factor 1), and its receptor IGF1R, inducing PI3K–AKT (phosphoinositol3kinase - proteinkinaseB) signalization for gonadal and adrenal androgenic hormone synthesis. By the availability and binding of the hormone, the androgenic receptor (> AR, nuclear transcription factor) is activated, since the metabolic corepressor FoxO1 (forkhead box O1) is phosphorylated and subsequently is translocated to the cytoplasm. Likewise, mTORC1 (mechanistic target of rapamycin complex 1) kinase regulating lipogenesis is liberated too, from FoxO1 suppression. It means that IGF-1 through AR mediation stimulates cell proliferation leading to production of sebocytes synthesising sebum and oil, and also to production of keratinocytes making hairs. If imbalanced by hormonal, nutrition, or metabolic reasons, excessive cell proliferations result in failed discharge of sebum, oil, corneocytes, through the follicular canal and skin disorders without inflammation (e.g. blackheads) or with inflammation (e.g. pustules) appear (comedogenesis). The process is accelerated by microbial commensals, residents on skin surface anaerob Gram positive Propionibacterium acnes, aerob Gram positive Staphylococcus epidermidis, lipophilic basidiomycetous Malassezia furfur. These microbial accelerators exert their effects by enhancing expression of keratinocyte surface Toll-like receptors (TLR2 és TLR4), by increasing synthesis of keratinocyte MMP9 (Matrix Metallopoteinase 9) for progression of inflammation set around PSU. Active substance The tretinoin mechanism of action is not completely unfolded. Effects of tretinoin (all-trans-retinsav) manifest presumably through activities of retinoc acid receptors {steroid receptor superfamily > cell nuclear retinoic acid receptors (RARs = RARα, RARβ, RARγ), retinoic acid receptor heterodimers (RAR-RXR with RXR = RXRα or RXRβ or RXRγ} also involving corepressors NCOR1, NCOR2 and coactivators NCOA1, NCOA2, NCOA3, regulating structure and function of retinoic acid receptor heterodimers. According to its morphogenic feature, tretinoin enhances cell proliferation in the follicular epithelium (epithelial cell turnover), supports detachment and release of overproduced corneocytes from the follicles (comedolysis) and by repressing among others TLR2 expression (see above), it decreases inflammation around PSU . Options in topical treatment of Acne vulgaris (slight and medium appearance) Benzoyl-peroxide (BP): traditional antibacterial and comedolytic gel or cream, with efficiency in time suitable for preventing antibacterial resistance. Drawbacks: skin irritation, hypersensitivity reaction (contact dermatitis). BP in combination: BP + clindamycin // BP + erythromycin // BP + nadifloxacin // BP + glycolyc acid // BP + urea (formulation: wash, gel, cream). Drawbacks: developing hypersensitivity reaction and/or resistance. Antibiotics (AB): clindamycin, erythromycin (in gel for oily skin; in ointment or lotion for dry skin; in foam for trunk) bind to bacterial ribosomal 50S subunit and irreversibly inhibit protein synthesis; they have antiinflammatory effects. Drawbacks: developing resistance. AB in combination: AB + tretinoin Drawbacks: AB in use Sulfur, sodium-sulfacetamide (wash, lotion, cream, foam). Drawbacks: skin irritation. Azelaic acid: antimicrobial substance inhibiting protein synthesis (in gel), preferably in combinations of +BP/+antibiotics /+tretinoin. Drawbacks: as above. Synthetic retinoids: adapalene, tretinoin, tazarotene. In treatment of acne vulgaris, retinoids prove to be the most efficient and safe substances (comedolysis, antiinflammatories). Drawbacks: skin irritation. Solutions: Effective, new generation drug delivery systems ensuring minimum quantity active substance for stable and controlled release. "microsponge delivery system" is a polymer of porous microspheres in gel / cream / powder /.... formulations hydrogels micronized tretinoin/nanogel: oil-in-water emulsion with oil drop diameter < 200nm, including lipophilic tretinoin dissolved. Alternatives: cosmeceuticals as solutions for treatment?