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WO2021257630A2 - Drugs and methods for reducing body odor and sweat - Google Patents

Drugs and methods for reducing body odor and sweat Download PDF

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Publication number
WO2021257630A2
WO2021257630A2 PCT/US2021/037518 US2021037518W WO2021257630A2 WO 2021257630 A2 WO2021257630 A2 WO 2021257630A2 US 2021037518 W US2021037518 W US 2021037518W WO 2021257630 A2 WO2021257630 A2 WO 2021257630A2
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composition
oligonucleotides
effective amount
rna
nucleotides
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WO2021257630A4 (en
WO2021257630A3 (en
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Richelle CUTLER
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Cutler Richelle
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    • C—CHEMISTRY; METALLURGY
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/047—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates having two or more hydroxy groups, e.g. sorbitol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7125—Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/323—Chemical structure of the sugar modified ring structure
    • C12N2310/3231—Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA
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    • C12N2310/30—Chemical structure
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    • C12N2320/00—Applications; Uses
    • C12N2320/30—Special therapeutic applications
    • C12N2320/32—Special delivery means, e.g. tissue-specific

Definitions

  • Axillary sweating can cause embarrassing body odor and stain clothes. Clothes are most often laundered to remove the scent of body odor from axillary sweating. Weekly laundering of clothes is time- consuming, costs money and wears clothes out. In addition, clothes laundering releases microplastics from synthetic clothes into the wastewater. The microfibers are small enough to pass through wastewater treatment plants and end up in the oceans where they cause problems in marine organisms. Reducing axillary sweating could reduce laundering frequency, which could help preserve the ecosystem.
  • osmidrosis osmidrosis
  • hyperhidrosis axillary sweating
  • Amputees often report skin problems resulting from residual limb sweating in their prosthetic device. People can be so impacted by their body odor or excessive sweating that they elect to have Botox injections, laser treatment, or surgical resection of axillary sweat glands.
  • Deodorants or antiperspirants are used to prevent body odor and sweating. For maximum efficacy, deodorants usually require daily reapplication. Even so, for many people, deodorants and antiperspirants do not adequately prevent body odor and sweating. Deodorants contain chemicals that inhibit bacteria growth, whereas antiperspirants contain chemicals that block sweat ducts. Both deodorants and antiperspirants often contain fragrances to mask odors. These, and other chemicals in deodorants and antiperspirants, such as triclosan, and cyclotetrasiloxane, are known endocrine disruptors and can accumulate in the body over time, posing a health risk. Deodorants and antiperspirants can also cause skin irritation in many people.
  • Most antiperspirants contain aluminum salts.
  • Aluminum salts plug sweat ducts by acid-mediated precipitation of the aluminum salt with mucopolysaccharides and ductal epithelium damage.
  • CrystalTM is a popular deodorant composed of potassium alum, i.e. potassium aluminum sulfate.
  • potassium alum i.e. potassium aluminum sulfate.
  • Potassium alum, or its synthetic precursor bauxite must be mined and thus, its consumption has an environmental cost.
  • Glycopyrronium tosylate (Qbrexza TM ) was recently approved by the FDA for primary hyperhidrosis. Glycopyrronium tosylate is an anticholinergic that prevents the contraction of eccrine myoepithelium. Qbrexza significantly reduced sweat production in most subjects but requires a daily application and is not without side effects due to systemic anticholinergic effects, the most common including dry mouth, blurred vision, application site pain, nasopharyngitis, and dilated pupils.
  • Embodiments disclosed herein include methods to reduce body odor and sweat volume by topical application of compositions that reduce ABCC11 transporter activity and/or AQP5 channel water conduction.
  • the composition includes single- stranded RNA and DNA oligonucleotides that attenuate ABCC11 and AQP5 expression by targeting and disrupting ABCC11 and/or AQP5 mRNA expression.
  • oligonucleotide penetration and transdermal delivery methods.
  • Chemical penetration enhancers are included in the topical composition to aid in transdermal oligonucleotide penetration.
  • Transdermal delivery methods include using a cell-penetration peptide, iontophoresis, microneedle array, and lipid-oligonucleotide conjugates or complexes.
  • a composition is comprised of chemical compounds that inhibit ABCC11 transporter and / or AQP5 activity and may be used in combination with oligonucleotides.
  • One such compound tetramethylpyrazine (TMP) can reduce body odor by inhibiting ABCC11 transporter activity.
  • Another compound, 1,3- propanediol can reduce sweat volume by blocking AQP5 water channel conduction.
  • TMP tetramethylpyrazine
  • 1,3- propanediol can reduce sweat volume by blocking AQP5 water channel conduction.
  • a composition comprised of single-stranded oligonucleotides, TMP, and 1,3 -propanediol can be delivered in a topical application directly into the sweat ducts in a transappendageal route.
  • the axillae apocrine and apoeccrine glands secrete lipophilic anions that cutaneous bacterial enzymes convert to odorous compounds.
  • the protein ABCC11 secretes molecules out of myoepithelium cells into the lumina of the secretory duct.
  • the protein ABCC11 is an ATP-binding cassette transporter and is a member of a large family of proteins called multi drug resistant-associated proteins.
  • a sequence variant (rsl 7822931) in the gene ABCC11 at position 538 affects sweat gland secretion.
  • the 538G>A variant changes the 180th amino acid from glycine to arginine, which increases the proteasomal degradation of the protein. Thus, the 538G/G allele has a higher expression level. Artificially attenuating ABCC11 gene expression in a subject significantly reduces apocrine and apoeccrine gland secretion, and thereby reduces body odor.
  • Sweat volume is determined by the amount of water released from aquaporin 5 (AQP5) water channels in secretory epithelial cells.
  • AQP5 is the only known water channel localized to the luminal membrane of the sweat gland ducts.
  • Polymorphisms in the AQP5 gene can affect channel expression and regulation. For instance, the AQP5 promoter polymorphism 1364A/C decreases AQP5 expression and affects sweat output. Artificially reducing AQP5 water channel gene expression would decrease sweating and secretion of organic molecules.
  • oligonucleotide refers to a single-stranded sequence of nucleotides that can be either RNA or DNA.
  • Embodiments described herein include a method of reducing body odor and sweat volume in a subject by topical application of compositions that inhibit ABCC 11 and AQP5 activity.
  • Oligonucleotides can reduce ABCC11 and AQP5 activity by targeting complementary sequences in ABCC11 (SEQ NO. 1) and AQP5 (SEQ NO. 2) mRNA (or pre-mRNA) and attenuating mRNA expression.
  • mRNA target sequences are selected using design tools.
  • single-stranded oligonucleotides compared to double-stranded siRNA, are more suitable for transdermal applications because a larger size restricts penetration through the stratum comeum.
  • single- stranded oligonucleotides are not as immunogenic, and their amphiphilic nature enhances cellular uptake by macropinocytosis.
  • researchers have shown that topically applied single- stranded oligonucleotides can penetrate through skin layers, enter cells, and attenuate gene expression in the epidermis and dermis in ex vivo human skin and animal models.
  • Single-stranded oligonucleotides can be composed of ribonucleic acid (RNA) or deoxyribonucleic acids (DNA).
  • Ribonucleic acid bases are adenosine, uracil, cytosine, and guanine, while deoxyribonucleic acid bases are adenosine, thymine, cytosine, and guanine.
  • a hydroxyl group (-OH) is present on the ribose carbon in ribonucleic acid but is absent (-H) in deoxyribonucleic acid.
  • Gene expression can be silenced by introducing short single-stranded oligonucleotides complementary to target mRNA, which can initiate mRNA degradation by several different mechanisms.
  • Single-stranded oligonucleotides composed of RNA can induce mRNA degradation by binding to the enzyme Argonaute-2, simulating unwound siRNA in RNA interference.
  • Argonaute-2 and the single-stranded RNA binds and cleaves the complementary mRNA.
  • Single-stranded oligonucleotides (antisense oligonucleotides) composed of DNA binds to the complementary mRNA and the enzyme RNase HI is recruited to the DNA-mRNA hybrid, where it cleaves mRNA, releasing the oligonucleotide intact.
  • Complementary oligonucleotides can also simply bind complementary mRNA sequences in the 5’UTR to inhibit ribosomal assembly by steric hindrance or 5’cap assembly, preventing 5’ -cap-dependent translation.
  • mRNA can be cleaved by a ribozyme, wherein the oligonucleotide encodes a minimized Ml RNA catalytic subunit of the E. coli RNase P ribozyme linked to a guide sequence complementary to the mRNA target.
  • the mRNA degradation mechanism utilized by single-stranded oligonucleotides depends on the composition, i.e. RNA vs. DNA, and by oligonucleotide length, modifications, and target location.
  • oligonucleotide target mRNA sequences with minimal secondary structure, 30-50% GC content, and low homology to other sequences.
  • Techniques for selecting oligonucleotides are outlined in Technical Bulletin #506, "siRNA Design Guidelines,"; and by other web-based design tools at “BLOCK-iTTM RNAi Designer” by Thermo Fisher Scientific. Initial search parameters can include G/C contents between 35% and 55%.
  • the target sequence may be in the coding region or the 5' or 3' untranslated regions of the mRNAs.
  • Oligonucleotide gene silencing is tested by transfecting the cells expressing the target mRNA with test oligonucleotide and scrambled control followed by transcript expression assessment by Quantitative reverse transcript real-time PCR.
  • a method of reducing body odor by attenuating ABCC11 mRNA includes administering a topical application of a composition comprised of an effective amount of oligonucleotide having a length of 14 to 25 nucleotides and a pharmaceutically acceptable carrier.
  • the oligonucleotide selected from the group consisting of a region of at least 9 contiguous nucleotides having at least 80% sequence complementarity to the mRNA, corresponding to any one of SEQ ID N0:3-SEQ ID NO: 16.
  • Table I lists examples of mRNA target sequences for ABCC11. (Note: thymine represents uracil for mRNA).
  • a method of reducing sweat output by attenuating APQ5 mRNA (SEQ ID NO. 3; GenBank accession NM 001651) expression includes a topical application of a composition comprised of an effective amount of oligonucleotides having a length of 14 to 25 nucleotides and a pharmaceutically acceptable carrier.
  • the oligonucleotide selected from the group consisting of a region of at least 9 contiguous nucleotides having at least 80% sequence complementarity to the mRNA, corresponding to any one of SEQ ID N0:17-SEQ ID N0:28.
  • Table II. lists examples of mRNA target sequences for APQ5. (Note: thymine represents uracil for mRNA).
  • Oligonucleotide modifications may include a combination of the following: o locked nucleic acid (LNA), methylene bridge; a-L-LNA (a diastereomer of LNA) o 2 ’-(9-methyl, 2 ’ -O-m ethoxy ethyl , 2’ -fluorine o methyl phosphonate (methyl group attached to phosphate) o phosphorothioate linkage (PS) o phosphodiester linkage (PO) o phosphodiamidate morpholino oligomer o gapmers with terminal or mixed nucleic acid modification of 2’- oxygen and 4’ -carbon in the furanose ring o 2 , -fluoroarabinonucleic acid o amide linked nucleotides o peptide nucleic acid (PNA) o 5’ -phosphonate
  • LNA locked nucleic acid
  • PNA peptide nucleic acid
  • Oligonucleotide modifications can differ based on the intended degradation mechanism (RNase HI or Argonaute-2 or hindrance).
  • An example of oligonucleotide modifications for Argonaute-2 degradation comprises a 21 -nucleotide strand with a 5 ’phosphonate, alternating PS and PO linkages, and two 3 ’-terminal adenosines modified with T -(9-methyl or LNA.
  • oligonucleotide modifications for RNase HI degradation comprises a 14-nucleotide strand with three 5’ LNA nucleotides, followed by 9 contiguous unmodified nucleotides with phosphodiester linkage (gapmer) and two 3’ terminal 2’- ( -methyl or LNA.
  • a lower number of ribosomal translation complexes can increase mRNA degradation by allowing more time for RNase HI binding and cleavage. Although, this effect depends on the oligonucleotide target location. For instance, RNase HI efficiency is less dependent on translation rate in the 3 ’UTR regions, while more 5 ’ targeting oligonucleotides are most effective during lower translation rates. Translation of ribosomal machinery follows a circadian rhythm, which affects bulk mRNA translation rate. Thus, RNase HI efficacy can be affected by the diurnal timing of antisense oligonucleotide application unless 3’-mRNA targets are chosen.
  • One embodiment involves conjugating or complexing the oligonucleotide to a cell-penetrating peptide or lipophilic molecule, which can enhance both skin permeability and cell internalization.
  • a cell-penetrating peptide or lipophilic molecule which can enhance both skin permeability and cell internalization.
  • cell-penetrating peptides include the following: VP22, TAT, POD, polyarginine, penetratin, meganin, Pip6a, TD-1, GGL27, and SPACE. Conjugating oligonucleotides to the SPACE peptide resulted in delivery to the depth of the dermis from a topical application. Other conjugated or complexed cell-penetrating peptides could perform similarly.
  • Another embodiment of the invention is to deliver oligonucleotides in microvesicles, such as exosomes.
  • Clinical-grade exosomes can be produced in large batches from an immortalize mesenchymal cell line. Mesenchyme cells do not express major histocompatibility complex (MHC) proteins and are therefore less immunogenic.
  • Oligonucleotides can be loaded directly into the exosome production cell by transfection, squeezing, or direct liposome fusion. Alternatively, oligonucleotides can be loaded post-generation by electroporation. Exosomes can be isolated from culture supernatant by size exclusion chromatography for optimal scale, cost, and purity. For quality control, oligonucleotides can be quantified by RNA isolation, reverse transcription, and quantitative RT-PCR.
  • oligonucleotides are delivered conjugated to a lipid, or as lipid/peptide-complexes.
  • Lipophilic molecules conjugated to 5’ -end of oligonucleotides, with an optimal length carbon linker could include cholesterol, oleyl alcohol, lithocholic acid, and oleylamide of lithocholic acid, eicosapentaenoic (EPA), palmitoyl, pyrrolidininum, and docosahexaenoic acid (DHA), can aid in tissue distribution and reduce the inflammatory response.
  • EPA eicosapentaenoic
  • DHA docosahexaenoic acid
  • the oligonucleotides described herein are targeted for cellular internalization in the secretory duct epithelium.
  • a conjugated fatty acid could ensure that oligonucleotides have minimal release into the circulation.
  • Apocrine and apoeccrine sweat glands in the axilla have long ducts, with a diameter of approximately 70-100 pm that expand into a secretory coil.
  • the secretory coil of the apocrine and apoeccrine sweat gland is localized in the hypodermis at the subcutaneous fat interface, approximately between 0.65-2.78 mm from the skin surface.
  • a transepidermal approach may be necessary for oligonucleotides to reach the full extent of the secretory coil (glomerulus).
  • Optimal delivery could require physically penetrating the stratum comeum, which consists of 10- 25 layers of keratinocytes.
  • One embodiment is to deliver oligonucleotides through microneedles to enhance transdermal delivery.
  • a microneedle array breaks through the stratum comeum to painlessly dispense oligonucleotides directly into a subject’s epidermis.
  • Oligonucleotides can be delivered in hollow needles (MicronJet600®), coated onto the microneedles, or applied after creating microporations. Alternatively, oligonucleotides can be incorporated into dissolvable microneedles in a patch application.
  • Another embodiment is to combine oligonucleotides with ionic compounds that mask the nucleic acid negative charge, along with chemical permeation enhancers, such as fatty acids.
  • Equimolar synergistic combinations are known to enhance topical oligonucleotide delivery.
  • Synergistic combinations may include glycols (propylene glycol), triethanol amine, mefenamic acid, Aquaphor, surfactants (sodium dodecyl sulfate), phenyl piperazine, alcohols, water, fatty acids, fatty esters, lauric acid, geranic acid, choline, propylparaben, methylparaben, isopropyl myristate, glyceryl monostearate, hydroxyproyl methylcellulose, polyoxyl-40-stearate, short-chain alkanols, tricaprylin, n-methyl pyrrolidone, oleic acid, bomeol, benzyl dimethyl octyl ammonium, and lauroylcholine.
  • glycols propylene glycol
  • triethanol amine mefenamic acid
  • Aquaphor Aquaphor
  • surfactants sodium dodecyl sulfate
  • phenyl piperazine alcohols
  • pharmaceutical formulations comprise oligonucleotides, or salts thereof, of the invention up to 99% by weight mixed with a physiologically acceptable carrier medium such as water, buffer, saline, glycine, hyaluronic acid, mannitol, and permeation enhancers.
  • a physiologically acceptable carrier medium such as water, buffer, saline, glycine, hyaluronic acid, mannitol, and permeation enhancers.
  • Oligonucleotides are administered as, solutions, suspensions, cream, ointment, or emulsions.
  • the following are examples of possible formulations.
  • Oligonucleotides may only produce a 40% knockdown in ABCC11 and AQP5 expression, thus combining oligonucleotide treatment with chemical compounds that reduce ABCC11 and AQP5 activity can be beneficial.
  • Small molecule ABC transporter inhibitors block chemotherapy drug efflux to reduce resistance to cytotoxic cancer chemotherapy.
  • Tetramethylpyrazine (TMP) with formula C 8 H
  • TMP is isolated from the Chinese herb Ligusticum Chuanxiong Hort , which has been used in Chinese herbal medicine for 2000 vear researchers are currently testing the safety and efficacy of TMP in human trials for treating various diseases, which appear unrelated to ABC transporter inhibition. TMP formulated with enhancers readily penetrates human skin. A topical application of TMP has few safety concerns. Topical TMP has been proposed as 1) a fragrance in deodorants and antiperspirants and 2) for transdermal delivery for distribution to the brain and heart to prevent middle cerebral artery occlusion. It is unknown whether a deodorant or antiperspirant product was ever formulated with TMP, although if so, it was not formulated for skin penetration; thus, inadvertent function as described herein is unlikely. Transdermal delivery of TMP for distribution to the brain or heart used methods for transdermal flux at a location (chest) without apocrine glands, and thus sans ABCC11, at high TMP concentrations (250 mg over 20 cm 2 ).
  • PDO 1,3 -propanediol
  • another embodiment involves a method of reducing body odor and sweat output by topically applying a composition comprised of an effective amount of TMP and/ or PDO.
  • Blocking APQ5 water channel conductance will further reduce the delivery of apocrine secreted molecules to the skin surface.
  • the physiochemical properties of TMP and PDO predict good transdermal penetration.
  • oligonucleotides and the chemical compounds, TMP and PDO will reduce ABCC11 and AQP5 activity for a different duration.
  • Gene expression silencing could last from 1 week to 6 months, whereas chemical compounds could last from 1-7 days.
  • Oligonucleotide and chemical compounds have different physiochemical properties, and thus, require different permeation enhancers, carriers, and delivery methods.
  • Maximum body odor and sweat output reduction may require a two-step process involving a weekly or monthly oligonucleotides application, and a daily or weekly chemical compound application. Alternatively, an individual may choose to use one or the other composition for convenience.
  • a topical application of oligonucleotide and chemical compounds can reduce ABCC11 and AQP5 activity by entering sweat ducts in a transappendageal route, providing direct access to luminal myoepithelial and secretory epithelial cells.
  • Eccrine and apoeccrine ducts open at the skin surface and make up approximately 65% of the sweat glands.
  • most apocrine sweat gland ducts open into the hair follicle, thus removing the hair follicle at the root by waxing could increase access to the apocrine duct.
  • a direct transappendageal route would significantly reduce the possibility of any inflammatory response and preclinical studies support feasibility.
  • Another embodiment is to drive transdermal oligonucleotides in the transappendageal route using iontophoresis set at a constant anode current in combination with a penetration enhancer pretreatment.
  • iontophoresis set at a constant anode current in combination with a penetration enhancer pretreatment.
  • Embodiments described herein make use of oligonucleotides to reduce body odor and sweat output by attenuating ABCC11 transporter and AQP5 water channel mRNA expression.
  • Embodiments involve using an effective concentration of the chemical compounds TMP and PDO to reduce ABCC11 transporter and AQP5 water channel activity, and thereby reduce body odor and sweat output.
  • the embodiments include a method of preventing body odor and sweat output in a subject by topical application of a composition comprised of oligonucleotides and/ or chemical compounds that reduce ABCC11 and AQP5 activity.
  • Embodiments include known formulations and delivery methods that enhance oligonucleotide skin penetration.
  • the specificity of gene expression silencing by oligonucleotide targeting, along with the affinity of TMP and PDO for ABCC1 1 and AQP5, respectively, has the potential to significantly reduced body odor and sweating free from anticholinergic side-effects.
  • Embodiments of the invention provide a solution to the longstanding need for a convenient, robust, painless, and long-lasting treatment to reduce body odor and sweating.
  • Embodiments of the invention have the potential to provide significant relief to those suffering from osmidrosis, bromhidrosis, or hyperhidrosis.
  • widespread use of this invention has the potential to reduce synthetic clothes laundering and thereby reduce the amount of microplastic fibers entering the ecosystem.

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Abstract

The ABC transporter, ABCCll, is expressed in secretory sweat duct myoepithelium and is responsible for secreting compounds that bacterial enzymes convert into odorous products. The Aquaporin 5 water channel is expressed in the secretory epithelium and secretes water into the sweat duct. Body odor and sweat production can be reduced by attenuating ABCC11 and Aquaporin 5 gene expression in human skin by topical application of a composition comprised of an effective amount of oligonucleotides having complementary sequences to ABCC11 and Aquaporin 5 mRNA. Embodiments of the invention include methods to enhance the delivery of the oligonucleotides using permeation enhancers, cell-penetrating peptides, lipid-oligonucleotide conjugates, microneedle array, and iontophoresis. The invention further describes reducing human body odor and sweat production by inhibiting ABCC11 and Aquaporin 5 activity by topical application of compositions comprised of an effective amount of tetramethylpyrazine and 1,3 -propanediol.

Description

DRUGS AND METHODS FOR REDUCING BODY ODOR AND
SWEAT
[0001] This application claims priority to U.S. Provisional Application No. 63/039,457, filed on June 15, 2020, the entire contents of which are incorporated herein by reference.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on June 15, 2021, is named 1000B-PCT.txt and is 19 kb in size.
BACKGROUND
[0003] Axillary sweating can cause embarrassing body odor and stain clothes. Clothes are most often laundered to remove the scent of body odor from axillary sweating. Weekly laundering of clothes is time- consuming, costs money and wears clothes out. In addition, clothes laundering releases microplastics from synthetic clothes into the wastewater. The microfibers are small enough to pass through wastewater treatment plants and end up in the oceans where they cause problems in marine organisms. Reducing axillary sweating could reduce laundering frequency, which could help preserve the ecosystem.
[0004] Some people have offensive body odor, known as osmidrosis, or profuse axillary sweating, known as hyperhidrosis. Amputees often report skin problems resulting from residual limb sweating in their prosthetic device. People can be so impacted by their body odor or excessive sweating that they elect to have Botox injections, laser treatment, or surgical resection of axillary sweat glands.
[0005] Deodorants or antiperspirants are used to prevent body odor and sweating. For maximum efficacy, deodorants usually require daily reapplication. Even so, for many people, deodorants and antiperspirants do not adequately prevent body odor and sweating. Deodorants contain chemicals that inhibit bacteria growth, whereas antiperspirants contain chemicals that block sweat ducts. Both deodorants and antiperspirants often contain fragrances to mask odors. These, and other chemicals in deodorants and antiperspirants, such as triclosan, and cyclotetrasiloxane, are known endocrine disruptors and can accumulate in the body over time, posing a health risk. Deodorants and antiperspirants can also cause skin irritation in many people.
[0006] Most antiperspirants contain aluminum salts. Aluminum salts plug sweat ducts by acid-mediated precipitation of the aluminum salt with mucopolysaccharides and ductal epithelium damage. Crystal™ is a popular deodorant composed of potassium alum, i.e. potassium aluminum sulfate. Many people suspect that the aluminum in antiperspirants absorbs into the body and can lead to toxic accumulation in the brain, which may contribute to a neurodegenerative process. Potassium alum, or its synthetic precursor bauxite, must be mined and thus, its consumption has an environmental cost.
[0007] Glycopyrronium tosylate (Qbrexza™) was recently approved by the FDA for primary hyperhidrosis. Glycopyrronium tosylate is an anticholinergic that prevents the contraction of eccrine myoepithelium. Qbrexza significantly reduced sweat production in most subjects but requires a daily application and is not without side effects due to systemic anticholinergic effects, the most common including dry mouth, blurred vision, application site pain, nasopharyngitis, and dilated pupils.
SUMMARY
[0008] Embodiments disclosed herein include methods to reduce body odor and sweat volume by topical application of compositions that reduce ABCC11 transporter activity and/or AQP5 channel water conduction. In one embodiment, the composition includes single- stranded RNA and DNA oligonucleotides that attenuate ABCC11 and AQP5 expression by targeting and disrupting ABCC11 and/or AQP5 mRNA expression.
[0009] Several embodiments pertain to formulations that enhance oligonucleotide penetration and transdermal delivery methods. Chemical penetration enhancers are included in the topical composition to aid in transdermal oligonucleotide penetration. Transdermal delivery methods include using a cell-penetration peptide, iontophoresis, microneedle array, and lipid-oligonucleotide conjugates or complexes.
[0010] In another embodiment, a composition is comprised of chemical compounds that inhibit ABCC11 transporter and / or AQP5 activity and may be used in combination with oligonucleotides. One such compound, tetramethylpyrazine (TMP) can reduce body odor by inhibiting ABCC11 transporter activity. Another compound, 1,3- propanediol, can reduce sweat volume by blocking AQP5 water channel conduction. A composition comprised of single-stranded oligonucleotides, TMP, and 1,3 -propanediol can be delivered in a topical application directly into the sweat ducts in a transappendageal route.
DETAILED DESCRIPTION
[0011] The axillae apocrine and apoeccrine glands secrete lipophilic anions that cutaneous bacterial enzymes convert to odorous compounds. The protein ABCC11 secretes molecules out of myoepithelium cells into the lumina of the secretory duct. The protein ABCC11 is an ATP-binding cassette transporter and is a member of a large family of proteins called multi drug resistant-associated proteins. A sequence variant (rsl 7822931) in the gene ABCC11 at position 538 affects sweat gland secretion. The 538G>A variant changes the 180th amino acid from glycine to arginine, which increases the proteasomal degradation of the protein. Thus, the 538G/G allele has a higher expression level. Artificially attenuating ABCC11 gene expression in a subject significantly reduces apocrine and apoeccrine gland secretion, and thereby reduces body odor.
[0012] Sweat volume is determined by the amount of water released from aquaporin 5 (AQP5) water channels in secretory epithelial cells. AQP5 is the only known water channel localized to the luminal membrane of the sweat gland ducts. Polymorphisms in the AQP5 gene can affect channel expression and regulation. For instance, the AQP5 promoter polymorphism 1364A/C decreases AQP5 expression and affects sweat output. Artificially reducing AQP5 water channel gene expression would decrease sweating and secretion of organic molecules. [0013] All the embodiments disclosed herein can be made and executed without undue experimentation and those skilled in the art will find the applied embodiments fully supported. Regardless, the embodiments should not be construed to unduly narrow the full scope of protection to which the present invention is entitled. The phrase, "attenuating” mRNA expression as used herein, means to reduce translation of the target mRNA into protein, through administering a number of oligonucleotides, resulting in mRNA cleavage by RNase HI or Argonaute-2, or through direct steric inhibition of translation machinery. The term “oligonucleotide” used herein, refers to a single-stranded sequence of nucleotides that can be either RNA or DNA.
[0014] Embodiments described herein include a method of reducing body odor and sweat volume in a subject by topical application of compositions that inhibit ABCC 11 and AQP5 activity. Oligonucleotides can reduce ABCC11 and AQP5 activity by targeting complementary sequences in ABCC11 (SEQ NO. 1) and AQP5 (SEQ NO. 2) mRNA (or pre-mRNA) and attenuating mRNA expression. In embodiments of the present invention, mRNA target sequences are selected using design tools.
[0015] The smaller size of single-stranded oligonucleotides, compared to double-stranded siRNA, are more suitable for transdermal applications because a larger size restricts penetration through the stratum comeum. Further, single- stranded oligonucleotides are not as immunogenic, and their amphiphilic nature enhances cellular uptake by macropinocytosis. Researchers have shown that topically applied single- stranded oligonucleotides can penetrate through skin layers, enter cells, and attenuate gene expression in the epidermis and dermis in ex vivo human skin and animal models. Moreover, new developments, such as cell-penetrating peptides, synergic penetration enhancers, iontophoresis, lipid conjugates or as lipid/peptide-complexes, and microneedle array, increase the efficiency of topical single-stranded oligonucleotide delivery [0016] Single-stranded oligonucleotides can be composed of ribonucleic acid (RNA) or deoxyribonucleic acids (DNA). Ribonucleic acid bases are adenosine, uracil, cytosine, and guanine, while deoxyribonucleic acid bases are adenosine, thymine, cytosine, and guanine. A hydroxyl group (-OH) is present on the ribose carbon in ribonucleic acid but is absent (-H) in deoxyribonucleic acid. Gene expression can be silenced by introducing short single-stranded oligonucleotides complementary to target mRNA, which can initiate mRNA degradation by several different mechanisms.
[0017] Single-stranded oligonucleotides composed of RNA (single- stranded- siRNA) can induce mRNA degradation by binding to the enzyme Argonaute-2, simulating unwound siRNA in RNA interference. Argonaute-2 and the single-stranded RNA binds and cleaves the complementary mRNA. Single-stranded oligonucleotides (antisense oligonucleotides) composed of DNA binds to the complementary mRNA and the enzyme RNase HI is recruited to the DNA-mRNA hybrid, where it cleaves mRNA, releasing the oligonucleotide intact. Complementary oligonucleotides can also simply bind complementary mRNA sequences in the 5’UTR to inhibit ribosomal assembly by steric hindrance or 5’cap assembly, preventing 5’ -cap-dependent translation. Lastly, mRNA can be cleaved by a ribozyme, wherein the oligonucleotide encodes a minimized Ml RNA catalytic subunit of the E. coli RNase P ribozyme linked to a guide sequence complementary to the mRNA target. The mRNA degradation mechanism utilized by single-stranded oligonucleotides depends on the composition, i.e. RNA vs. DNA, and by oligonucleotide length, modifications, and target location.
[0018] There are similar guidelines for selecting target mRNA with different degradation mechanisms. For instance, optimal oligonucleotide target mRNA sequences with minimal secondary structure, 30-50% GC content, and low homology to other sequences. Techniques for selecting oligonucleotides are outlined in Technical Bulletin #506, "siRNA Design Guidelines,"; and by other web-based design tools at “BLOCK-iT™ RNAi Designer” by Thermo Fisher Scientific. Initial search parameters can include G/C contents between 35% and 55%. The target sequence may be in the coding region or the 5' or 3' untranslated regions of the mRNAs. Oligonucleotide gene silencing is tested by transfecting the cells expressing the target mRNA with test oligonucleotide and scrambled control followed by transcript expression assessment by Quantitative reverse transcript real-time PCR.
[0019] One embodiment, a method of reducing body odor by attenuating ABCC11 mRNA (SEQ ID no. 1; GenBank accession AF367202.1) expression includes administering a topical application of a composition comprised of an effective amount of oligonucleotide having a length of 14 to 25 nucleotides and a pharmaceutically acceptable carrier. The oligonucleotide selected from the group consisting of a region of at least 9 contiguous nucleotides having at least 80% sequence complementarity to the mRNA, corresponding to any one of SEQ ID N0:3-SEQ ID NO: 16. Table I. lists examples of mRNA target sequences for ABCC11. (Note: thymine represents uracil for mRNA).
Figure imgf000011_0001
Number ABCC 11 starting Region
Figure imgf000011_0002
Target sequence nucleotide
3 GCTGAAAGAATTGGCAGGAACTGAA 53 5TJTR
4 GGGAAGTATGATGCTGCCTTGAGAA 262 Coding
5 CATCTGCTTCTGCATTGCCAGTGTA 588 Coding
6 CCGGTGATGTAAACTACCTGTTTGA 872 Coding
7 TGCAGCATTTCTTCCTACTTCATTA 949 Coding
8 CACCAGCTGCAGTACTTAGAATT 2148 Coding
9 CATGCTGCAGGACACAGCAAAGATA 2298 Coding
10 AAGAAGGCTCCTTGAGTTGGAGG 2428 Coding
11 CGGCCCTGCACAACAAGCTCTTTAA 2756 Coding
12 GAGCAGTTCCTGGTCCTGTCCTTAA 2893 Coding
13 AACCAU CAGAUU C AACCUAGAU C 3767 Coding
14 AAGAU CAU CCUUAU CGAUGAAGC 3966 Coding
15 CAATGGGAAGGTGGTAGAATTTGAT 4125 Coding
16 GGAAACCCTGGAATAGGCTACTTGA 4380 3TJTR
[0020] In another embodiment, a method of reducing sweat output by attenuating APQ5 mRNA (SEQ ID NO. 3; GenBank accession NM 001651) expression includes a topical application of a composition comprised of an effective amount of oligonucleotides having a length of 14 to 25 nucleotides and a pharmaceutically acceptable carrier. The oligonucleotide selected from the group consisting of a region of at least 9 contiguous nucleotides having at least 80% sequence complementarity to the mRNA, corresponding to any one of SEQ ID N0:17-SEQ ID N0:28. Table II. lists examples of mRNA target sequences for APQ5. (Note: thymine represents uracil for mRNA).
Figure imgf000012_0001
Number Target sequence starting Region nucleotide
Figure imgf000012_0002
TTCTTGGCCACCCTCATCTT 193 Coding
18 TGGCCACCCTCATCTTCGTCTTCTT 202 Coding
19 GGCCACCCTCATCTTCGTCTTCTTT 203 Coding
20 GCCACCCTCATCTTCGTCTTCTTTG 204 Coding
21 CCACCCTCATCTTCGTCTTCTTTGG 205 Coding
22 TGGTGGAGCTGATTCTGACCTTCCA 544 Coding
23 GAGTGAGCGTGTGGCCATCATCAAA 842 Coding
24 AGTGAGCGTGTGGCCATCATCAAAG 843 Coding
25 AAGTACCTAACACAAGCTTCCTT 964 3’ UTR
26 CAGAGATTGTGMTGCAGTGCCAAG 1174 3’ UTR
27 GATTGTGMTGCAGTGCCAAGCTCA 1178 3’ UTR
28 AAGACAGACUGGUUCAUUGAAUG 1340 3’UTR
[0021] Oligonucleotide modifications may include a combination of the following: o locked nucleic acid (LNA), methylene bridge; a-L-LNA (a diastereomer of LNA) o 2 ’-(9-methyl, 2 ’ -O-m ethoxy ethyl , 2’ -fluorine o methyl phosphonate (methyl group attached to phosphate) o phosphorothioate linkage (PS) o phosphodiester linkage (PO) o phosphodiamidate morpholino oligomer o gapmers with terminal or mixed nucleic acid modification of 2’- oxygen and 4’ -carbon in the furanose ring o 2,-fluoroarabinonucleic acid o amide linked nucleotides o peptide nucleic acid (PNA) o 5’ -phosphonate
[0022] Oligonucleotide modifications can differ based on the intended degradation mechanism (RNase HI or Argonaute-2 or hindrance). An example of oligonucleotide modifications for Argonaute-2 degradation (ss-siRNA) comprises a 21 -nucleotide strand with a 5 ’phosphonate, alternating PS and PO linkages, and two 3 ’-terminal adenosines modified with T -(9-methyl or LNA.
[0023] An example of oligonucleotide modifications for RNase HI degradation (antisense oligonucleotides) comprises a 14-nucleotide strand with three 5’ LNA nucleotides, followed by 9 contiguous unmodified nucleotides with phosphodiester linkage (gapmer) and two 3’ terminal 2’- ( -methyl or LNA.
[0024] A lower number of ribosomal translation complexes can increase mRNA degradation by allowing more time for RNase HI binding and cleavage. Although, this effect depends on the oligonucleotide target location. For instance, RNase HI efficiency is less dependent on translation rate in the 3 ’UTR regions, while more 5 ’ targeting oligonucleotides are most effective during lower translation rates. Translation of ribosomal machinery follows a circadian rhythm, which affects bulk mRNA translation rate. Thus, RNase HI efficacy can be affected by the diurnal timing of antisense oligonucleotide application unless 3’-mRNA targets are chosen.
[0025] One embodiment involves conjugating or complexing the oligonucleotide to a cell-penetrating peptide or lipophilic molecule, which can enhance both skin permeability and cell internalization. There are several known cell-penetrating peptides, all of which feature cationic amino acids, such as lysine or arginine. Some known cell-penetrating peptides include the following: VP22, TAT, POD, polyarginine, penetratin, meganin, Pip6a, TD-1, GGL27, and SPACE. Conjugating oligonucleotides to the SPACE peptide resulted in delivery to the depth of the dermis from a topical application. Other conjugated or complexed cell-penetrating peptides could perform similarly.
[0026] Another embodiment of the invention is to deliver oligonucleotides in microvesicles, such as exosomes. Clinical-grade exosomes can be produced in large batches from an immortalize mesenchymal cell line. Mesenchyme cells do not express major histocompatibility complex (MHC) proteins and are therefore less immunogenic. Oligonucleotides can be loaded directly into the exosome production cell by transfection, squeezing, or direct liposome fusion. Alternatively, oligonucleotides can be loaded post-generation by electroporation. Exosomes can be isolated from culture supernatant by size exclusion chromatography for optimal scale, cost, and purity. For quality control, oligonucleotides can be quantified by RNA isolation, reverse transcription, and quantitative RT-PCR.
[0027] In another embodiment, oligonucleotides are delivered conjugated to a lipid, or as lipid/peptide-complexes. Lipophilic molecules conjugated to 5’ -end of oligonucleotides, with an optimal length carbon linker, could include cholesterol, oleyl alcohol, lithocholic acid, and oleylamide of lithocholic acid, eicosapentaenoic (EPA), palmitoyl, pyrrolidininum, and docosahexaenoic acid (DHA), can aid in tissue distribution and reduce the inflammatory response. Unlike topical RNA applications aiming for transdermal passage, the oligonucleotides described herein are targeted for cellular internalization in the secretory duct epithelium. A conjugated fatty acid could ensure that oligonucleotides have minimal release into the circulation.
[0028] Apocrine and apoeccrine sweat glands in the axilla have long ducts, with a diameter of approximately 70-100 pm that expand into a secretory coil. The secretory coil of the apocrine and apoeccrine sweat gland is localized in the hypodermis at the subcutaneous fat interface, approximately between 0.65-2.78 mm from the skin surface. A transepidermal approach may be necessary for oligonucleotides to reach the full extent of the secretory coil (glomerulus). Optimal delivery could require physically penetrating the stratum comeum, which consists of 10- 25 layers of keratinocytes.
[0029] One embodiment is to deliver oligonucleotides through microneedles to enhance transdermal delivery. A microneedle array breaks through the stratum comeum to painlessly dispense oligonucleotides directly into a subject’s epidermis. Oligonucleotides can be delivered in hollow needles (MicronJet600®), coated onto the microneedles, or applied after creating microporations. Alternatively, oligonucleotides can be incorporated into dissolvable microneedles in a patch application.
[0030] Another embodiment is to combine oligonucleotides with ionic compounds that mask the nucleic acid negative charge, along with chemical permeation enhancers, such as fatty acids. Equimolar synergistic combinations are known to enhance topical oligonucleotide delivery. Synergistic combinations may include glycols (propylene glycol), triethanol amine, mefenamic acid, Aquaphor, surfactants (sodium dodecyl sulfate), phenyl piperazine, alcohols, water, fatty acids, fatty esters, lauric acid, geranic acid, choline, propylparaben, methylparaben, isopropyl myristate, glyceryl monostearate, hydroxyproyl methylcellulose, polyoxyl-40-stearate, short-chain alkanols, tricaprylin, n-methyl pyrrolidone, oleic acid, bomeol, benzyl dimethyl octyl ammonium, and lauroylcholine.
[0031 ] In some embodiments, pharmaceutical formulations comprise oligonucleotides, or salts thereof, of the invention up to 99% by weight mixed with a physiologically acceptable carrier medium such as water, buffer, saline, glycine, hyaluronic acid, mannitol, and permeation enhancers. Oligonucleotides are administered as, solutions, suspensions, cream, ointment, or emulsions. The following are examples of possible formulations.
Formula _ Amount in weight, %
Polyoxyl-40-stearate 15
Hy droxyproyl 0.5 methycellulose
Glyceryl monostearate 10
Isopropyl myristate 10
Methylparaben 0.5
Propylparaben 0.5
RNase-free water 58.5
Oligonucleotides_ 5
Formula Amount in weight, %
Sodium laureth sulfate 10
Phenyl piperazine 10
RNase-free water 75
Oligonucleotides 5
[0032] Oligonucleotides may only produce a 40% knockdown in ABCC11 and AQP5 expression, thus combining oligonucleotide treatment with chemical compounds that reduce ABCC11 and AQP5 activity can be beneficial. Small molecule ABC transporter inhibitors block chemotherapy drug efflux to reduce resistance to cytotoxic cancer chemotherapy. Tetramethylpyrazine (TMP), with formula C8H|2N2. is a small polar bioactive that can inhibit ABCC transporters 1, 2, 3, 5. TMP is also likely to inhibit ABCCl 1 considering its sequence similarity with ABCC5. TMP is isolated from the Chinese herb Ligusticum Chuanxiong Hort , which has been used in Chinese herbal medicine for 2000 vear Researchers are currently testing the safety and efficacy of TMP in human trials for treating various diseases, which appear unrelated to ABC transporter inhibition. TMP formulated with enhancers readily penetrates human skin. A topical application of TMP has few safety concerns. Topical TMP has been proposed as 1) a fragrance in deodorants and antiperspirants and 2) for transdermal delivery for distribution to the brain and heart to prevent middle cerebral artery occlusion. It is unknown whether a deodorant or antiperspirant product was ever formulated with TMP, although if so, it was not formulated for skin penetration; thus, inadvertent function as described herein is unlikely. Transdermal delivery of TMP for distribution to the brain or heart used methods for transdermal flux at a location (chest) without apocrine glands, and thus sans ABCC11, at high TMP concentrations (250 mg over 20 cm2).
[0033] The molecule 1,3 -propanediol (PDO) is predicted to fit deep into the AQP5 water channel and block the channel with an IC50 of 0.245 m M and a Kd of 0.3m The prediction is based on computational simulations with proven accuracy. PDO is an amphiphilic surfactant with the formula Cl CiCl hOi 1)2.
[0034] Accordingly, another embodiment involves a method of reducing body odor and sweat output by topically applying a composition comprised of an effective amount of TMP and/ or PDO. Blocking APQ5 water channel conductance will further reduce the delivery of apocrine secreted molecules to the skin surface. The physiochemical properties of TMP and PDO predict good transdermal penetration.
[0035] Treatment with oligonucleotides and the chemical compounds, TMP and PDO, will reduce ABCC11 and AQP5 activity for a different duration. Gene expression silencing could last from 1 week to 6 months, whereas chemical compounds could last from 1-7 days. Oligonucleotide and chemical compounds have different physiochemical properties, and thus, require different permeation enhancers, carriers, and delivery methods. Maximum body odor and sweat output reduction may require a two-step process involving a weekly or monthly oligonucleotides application, and a daily or weekly chemical compound application. Alternatively, an individual may choose to use one or the other composition for convenience.
[0036] In another embodiment, a topical application of oligonucleotide and chemical compounds can reduce ABCC11 and AQP5 activity by entering sweat ducts in a transappendageal route, providing direct access to luminal myoepithelial and secretory epithelial cells. Eccrine and apoeccrine ducts open at the skin surface and make up approximately 65% of the sweat glands. However, most apocrine sweat gland ducts open into the hair follicle, thus removing the hair follicle at the root by waxing could increase access to the apocrine duct. A direct transappendageal route would significantly reduce the possibility of any inflammatory response and preclinical studies support feasibility.
[0037] Another embodiment is to drive transdermal oligonucleotides in the transappendageal route using iontophoresis set at a constant anode current in combination with a penetration enhancer pretreatment. Below is a formulation example:
Iontophoresis _ Amount in weight, %
Phosphate-buffered saline Ethanol diluent 15 Limonene/70% ethanol 1:1 pretreatment 85 Oligonucleotide 99% Purified water RNase-free 1.0
[0038] Embodiments described herein make use of oligonucleotides to reduce body odor and sweat output by attenuating ABCC11 transporter and AQP5 water channel mRNA expression. Embodiments involve using an effective concentration of the chemical compounds TMP and PDO to reduce ABCC11 transporter and AQP5 water channel activity, and thereby reduce body odor and sweat output. CONCLUSION
[0039] The embodiments include a method of preventing body odor and sweat output in a subject by topical application of a composition comprised of oligonucleotides and/ or chemical compounds that reduce ABCC11 and AQP5 activity. Embodiments include known formulations and delivery methods that enhance oligonucleotide skin penetration. [0040] The specificity of gene expression silencing by oligonucleotide targeting, along with the affinity of TMP and PDO for ABCC1 1 and AQP5, respectively, has the potential to significantly reduced body odor and sweating free from anticholinergic side-effects. Embodiments of the invention provide a solution to the longstanding need for a convenient, robust, painless, and long-lasting treatment to reduce body odor and sweating. Embodiments of the invention have the potential to provide significant relief to those suffering from osmidrosis, bromhidrosis, or hyperhidrosis. In addition, widespread use of this invention has the potential to reduce synthetic clothes laundering and thereby reduce the amount of microplastic fibers entering the ecosystem.

Claims

1. A composition of RNA or DNA antisense oligonucleotides comprised of a pharmaceutically acceptable carrier and oligonucleotides 13-25 nucleotides in length with 80% contiguous sequence similarity to oligonucleotides selected from the group consisting of SEQ ID NO. 3-27.
2. A composition according to claim 1, wherein the composition is further comprised of permeation enhancers.
3. A composition according to claim 1 , wherein oligonucleotide modifications are selected from the group consisting of locked nucleic acid (LNA), methylene bridge; a-L-LNA (a diastereomer of LNA), 2,-0-methyl, 2’-0- m ethoxy ethyl, -fluorine, methyl phosphonate (methyl group attached to phosphate), phosphorothioate linkage (PS), phosphodiester linkage (PO), phosphodiamidate morpholino oligomer, gapmers with terminal or mixed nucleic acid modification of -oxygen and 4’ -carbon in the furanose ring, 2,-fluoroarabinonucleic acid, amide linked nucleotides, peptide nucleic acid (PNA), and 5’ -phosphonate.
4. A method for reducing human body odor by administering a topical application of a composition comprised an effective amount of single- stranded RNA or DNA oligonucleotides having the length of 13-25 nucleotides and a pharmaceutically acceptable carrier, wherein the oligonucleotide sequence is 80% complementary to any contiguous ABCC1 1 mRNA sequence (SEQ ID. NO 1), and wherein the oligonucleotides selectively inhibit ABCC11 protein expression.
5. A method according to claim 4, wherein the RNA or DNA oligonucleotide composition is composed of 13-25 nucleotides with 80% contiguous sequence similarity to oligonucleotides selected from the group consisting of SEQ ID NO. 3-16.
6. A method according to claim 4, wherein the RNA or DNA composition is further comprised of permeation enhancers.
7. A method according to claim 4, wherein the RNA or DNA oligonucleotides have modifications selected from the group consisting of locked nucleic acid (LNA), methylene bridge; a-L-LNA (a diastereomer of LNA), 2,-0-methyl, 2,-0-methoxyethyl, 2,-fluorine, methyl phosphonate (methyl group attached to phosphate), phosphorothioate linkage (PS), phosphodiester linkage (PO), phosphodiamidate morpholino oligomer, gapmers with terminal or mixed nucleic acid modification of T -oxygen and 4’ -carbon in the furanose ring, 2,-fluoroarabinonucleic acid, amide linked nucleotides, peptide nucleic acid (PNA), and 5’ -phosphonate.
8. A method for reducing sweat by administering a topical application of a composition comprised of an effective amount of single-stranded RNA or DNA oligonucleotides and having the length of 13-25 nucleotides and a pharmaceutically acceptable carrier, wherein the oligonucleotide sequence is 80% complementary to any contiguous AQP5 mRNA sequence (SEQ ID. NO 2), wherein the oligonucleotides selectively inhibit AQP5 protein expression.
9. A method according to claim 8, wherein the composition of RNA or DNA oligonucleotides are 13-25 nucleotides in length with 80% contiguous sequence similarity to oligonucleotides selected from the group consisting of SEQ ID NO. 17-27.
10. A method according to claim 8, wherein the RNA or DNA composition is further comprised of permeation enhancers.
11. A method according to claim 8, wherein the RNA or DNA oligonucleotides have modifications selected from the group consisting of locked nucleic acid (LNA), methylene bridge; a-L-LNA (a diastereomer of LNA), 2,-0-methyl, 2,-0-methoxyethyl, 2,-fluorine, methyl phosphonate (methyl group attached to phosphate), phosphorothioate linkage (PS), phosphodiester linkage (PO), phosphodiamidate morpholino oligomer, gapmers with terminal or mixed nucleic acid modification of -oxygen and 4’ -carbon in the furanose ring, 2,-fluoroarabinonucleic acid, amide linked nucleotides, peptide nucleic acid (PNA), and 5’ -phosphonate.
12. A method according to claim 4, wherein the composition is further comprised of an effective amount of single- stranded RNA or DNA oligonucleotides with a length between 13-25 nucleotides and a pharmaceutically acceptable carrier, wherein the oligonucleotide contiguous sequence is 80% complementary to the AQP5 mRNA sequence (SEQ ID. NO 2), and wherein the oligonucleotide selectively inhibits AQP5 protein expression.
13. A method according to claim 12, wherein the composition of RNA or DNA oligonucleotides are 13-25 nucleotides in length with 80% contiguous sequence similarity to oligonucleotides selected from the group consisting of SEQ ID NO. 3-27.
14. A method according to claim 13, wherein the RNA or DNA composition is further comprised of permeation enhancers.
15. A method according to claim 13, wherein the composition is further comprised of an effective amount of tetramethylpyrazine.
16. A method according to claim 13, wherein the composition is further comprised of an effective amount of 1,3 -propanediol.
17. A method according to claim 13, wherein the composition is further comprised of an effective amount of tetramethylpyrazine and an effective amount of 1,3 -propanediol.
18. A method according to claim 14, wherein the composition is further comprised of an effective amount of 1,3 -propanediol.
19. A method according to claim 14, wherein the composition is further comprised of an effective amount of tetramethylpyrazine.
20. A method according to claim 14, wherein the composition is further comprised of an effective amount of tetramethylpyrazine and an effective amount of 1,3 -propanediol.
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