US20230414708A1 - Topical compositions comprising pea proteins and polyphenols - Google Patents

Topical compositions comprising pea proteins and polyphenols Download PDF

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US20230414708A1
US20230414708A1 US18/464,295 US202318464295A US2023414708A1 US 20230414708 A1 US20230414708 A1 US 20230414708A1 US 202318464295 A US202318464295 A US 202318464295A US 2023414708 A1 US2023414708 A1 US 2023414708A1
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acid
pea protein
polyphenol
disorders
fungal
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Miguel Angel Alonso Cohen
Marco Di Fulvio
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Devintec Sagl
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    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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Definitions

  • the object of the invention is topical compositions comprising pea proteins and polyphenols and the use thereof for the treatment of local or generalised disorders caused by increased permeation of the natural epithelial barriers.
  • Said disorders may be of bacterial, viral, inflammatory, allergic and/or fungal origin, or endogenous or idiopathic.
  • the complex of proanthocyanidins with pea protein has been proposed (WO2014154796). Said complex is particularly effective in the treatment of disorders caused by alterations of the intestinal epithelial tissue (non-ciliated simple columnar epithelium).
  • WO2018167131 describes combinations and conjugated products of pea protein and polysaccharides, in particular xyloglucans, fucoidans and ulvans, for topical treatment of inflammatory skin disorders such as atopic dermatitis, psoriasis, rosacea and rashes.
  • FIG. 1 A shows that pre-treatment at various time with formulations 1-3 preserved the reduction in TEER induced by Candida.
  • FIG. 1 B shows that pretreatment at various time with formulations 1-3 preserved the membrane integrity of fibroblasts infected with S. aureus.
  • FIG. 2 A shows that post-treatment increased TEER 6 and 24 hours after Candida infections.
  • FIG. 2 B shows that post-treatment preserved membrane integrity to a time dependent extent as from 6 hours after the S. aureus infection.
  • FIG. 3 A shows loss of membrane integrity in fibroblast infected by Candida when the mixtures were administered simultaneously with the infection.
  • FIG. 3 B shows limited increased TEER in fibroblast infected by S. aureus when the mixtures were administered simultaneous with the infection.
  • FIG. 4 A shows that pre-treatment at various time with formulations 1-3 preserved the membrane properties against Candida infection.
  • FIG. 4 B shows that pre-treatment at various time with formulations 1-3 preserved the membrane properties against Gardnerella vaginalis infection.
  • FIG. 5 A shows that post-treatment preserved membrane integrity at 6 hours after Candida infection.
  • FIG. 5 B shows that post-treatment preserved membrane integrity 24 hours after Candida infection.
  • FIGS. 6 A and 6 B show that when the mixtures were administered simultaneously with the infection, TEER increased in the infected cells.
  • pea protein and low-molecular-weight polyphenols belonging to the hydroxybenzoic and hydroxycinnamic acid families is particularly effective in the treatment of skin and mucosal disorders due to impairment of the barrier effect and of the intercellular tight junctions of the dermal and mucosal epithelia.
  • the invention therefore relates to topical formulations comprising combinations or complexes of pea protein with polyphenols belonging to the hydroxycinnamic and hydroxybenzoic acid families.
  • a further aspect of the invention relates to said combinations, complexes and formulations for use in the treatment of skin and mucosal disorders due to impairment of the barrier effect, and in particular in the treatment of local or generalised disorders caused by increased permeation of the natural epithelial barriers.
  • the formulations according to the invention exhibit increased specificity of action and optimum technological characteristics, which allow wide formulation flexibility.
  • skin and mucosal disorders due to impairment of the barrier effect is clear to the skilled person, as demonstrated, for example, by the following references: Grainne M. O. et al., “Filaggrin in atopic dermatitis”, J. Allergy Clin. Immunol., 2008, vol. 122(4), 689-693; Dorota Purzycka-Bohdan et al., “Genetic background of skin barrier dysfunction in the pathogenesis of psoriasis vulgaris”, Postepy Dermatol Allergol., 2015, 32(2), 123-126; and Flavia Alvim Sant'Anna Addor “Skin barrier in rosacea”, Ann. Bras Dermatol., 2016, vol. 91 (1), 59-63.
  • Atopic dermatitis, psoriasis, rosacea, contact dermatitis, cutaneous candidiasis, fungal or bacterial infections are examples of disorders wherein the barrier effect of the skin is impaired.
  • Vaginal candidiasis is an example of a disorder wherein the barrier effect of the mucosa is impaired.
  • compositions according to the invention facilitate inhibition of the mechanisms whereby allergens, bacteria, viruses and fungi pass through the epithelia, by reducing the permeability of the tight junctions and inhibiting paracellular transport to other systems.
  • the compositions of the invention also control and reduce water loss, with a consequent reduction in exposure to pro-allergenic factors.
  • the reinstatement of the barrier effect or prevention of its impairment obtainable with the formulations according to the invention also effectively counteract the transfer of the disorder from the epithelium to other systems, a phenomenon known as the “atopic march”, described by Selene K. Bantz et al., “ The Atopic March: Progression from Atopic Dermatitis to Allergic Rhinitis and Asthma ” in J Clin Cell Immunol. 2014 April; 5(2): 202.
  • Pea protein here means a protein powder containing legumin obtained by extraction by known methods from Pisum sativum seeds. Pea protein is available on the market under the trademarks Nutralys®, Pisane® and P80X. Commercially available products can be obtained, for example, from Cosucra and Dal Cin Gildo S.p.a. The product presents as a yellowish powder with a characteristic odour, which is poorly soluble in water.
  • the polyphenols which can be used according to the invention belong to the class of hydroxycinnamic and hydroxybenzoic acids.
  • Examples of said polyphenols comprise ferulic acid, isoferulic acid, chlorogenic acid, ellagic acid, gallic acid and rosmarinic acid, preferably ellagic acid and chlorogenic acid.
  • the polyphenols can be present in a substantially pure, isolated form, or in the form of extracts containing them in percentages ranging from 10 to 80%, depending on the source.
  • a known source of ellagic acid for example, is pomegranate extract, which has an ellagic acid content typically ranging between 30 and 60% by weight.
  • a known source of chlorogenic acid for example, is green coffee extract, which has a chlorogenic acid content typically ranging between 35 and 65% by weight.
  • the weight ratio of polyphenol to pea proteins ranges between 0.4:1 and 1:10; the ratio preferably ranges from 1:1 to 1:6.
  • the weight ratio of pea protein to an extract containing 40 to 50% by weight of polyphenol can range from 1:1 to 1:3, preferably 1:1.
  • suitable forms of administration comprise ointments, lotions, creams, solutions, suspensions, gels, mouthwashes, medicated patches or gauze, powders, pessaries and other forms suitable for topical application to the skin and mucosa.
  • the weight percentage of the pea protein can range from 1 to 30% of the total weight of the formulation, preferably from 1.5 to 10%, while the weight percentage of the polyphenol can range from 0.5 to 25% of the total weight of the formulation.
  • the formulations of the invention are prepared by known methods and with known carriers.
  • the mixture of polyphenol and pea protein according to the invention is preferably associated with moisturising agents, emollients, humectants, preservatives, antioxidants and pH regulators. Examples of said agents are given in WO 2018/167131.
  • Non-lipidic acidifiers such as lectin, lactic acid, sericin, poloxamers, carrageenan and arabinoxylan are particularly preferred. Of these, the following are particularly preferred: one or more of the following compounds selected from lectin, poloxamers, carrageenan and arabinoxylan, each at concentrations ranging from 0.1 to 2.5% by weight.
  • the formulations according to the invention can contain other active ingredients such as sucralfate, hyaluronic acid, antibiotics, steroidal and non-steroidal anti-inflammatories, chitosan, antihistamines and extracts of medicinal plants, in particular Zanthoxylum bungeanum , Aloe vera and camomile extracts.
  • active ingredients such as sucralfate, hyaluronic acid, antibiotics, steroidal and non-steroidal anti-inflammatories, chitosan, antihistamines and extracts of medicinal plants, in particular Zanthoxylum bungeanum , Aloe vera and camomile extracts.
  • compositions according to the invention are particularly useful for the treatment of atopic dermatitis, rosacea, psoriasis, contact dermatitis, dermatitis caused by exposure to sunlight, radiotherapy or chemotherapy, cutaneous candidiasis, fungal infections, vaginal candidiasis, polybacterial vaginitis, fungal and bacterial infections of the skin and mucosa, and infections caused by protozoa.
  • Pre-treatment at various times with formulations 1-3 preserved the reduction in TEER induced by Candida ( FIG. 1 A ) and the membrane integrity of fibroblasts infected with S. aureus ( FIG. 1 B ).
  • Post-treatment increased TEER 6 and 24 hours after the Candida infection ( FIG. 2 A ) and preserved membrane integrity to a time-dependent extent as from 6 hours after the S. aureus infection ( FIG. 2 B ).
  • Pre-treatment at various times with formulations 1-3 preserved the membrane properties against CA and GV infection ( FIGS. 4 A and 4 B ).
  • TEER increased in the infected cells ( FIGS. 6 A and 6 B ).

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Abstract

The object of the invention is topical compositions comprising pea proteins and polyphenols and the use thereof for the treatment of local or generalised disorders caused by increased permeation of the natural epithelial barriers. Said disorders may be of bacterial, viral, inflammatory, allergic and/or fungal origin, or endogenous or idiopathic.

Description

  • This application is a Divisional Application of U.S. application Ser. No. 17/757,832 filed Jun. 22, 2022, which is a U.S. national stage of PCT/EP2020/087490 filed 21 Dec. 2020, which claims priority to and the benefit of Italian Patent Application No. 102019000025246 filed 23 Dec. 2019, the contents of which are incorporated herein by reference in their entireties.
  • The object of the invention is topical compositions comprising pea proteins and polyphenols and the use thereof for the treatment of local or generalised disorders caused by increased permeation of the natural epithelial barriers. Said disorders may be of bacterial, viral, inflammatory, allergic and/or fungal origin, or endogenous or idiopathic.
    • PRIOR ART
  • Complexes of tannins with animal proteins, especially gelatin of bovine origin, albumin, casein or ovalbumin, have been proposed as effective remedies for gastrointestinal disorders.
  • The use of said complexes in the treatment of diarrhoea, for example, is disclosed in EP 1764105, EP 2526939, EP 2361623 and US 20090062191. Gelatin tannate has been available on the market for some time as a medical device for the treatment of acute diarrhoea.
  • As an alternative to the use of the tannin and animal protein complexes, the complex of proanthocyanidins with pea protein has been proposed (WO2014154796). Said complex is particularly effective in the treatment of disorders caused by alterations of the intestinal epithelial tissue (non-ciliated simple columnar epithelium).
  • WO2018167131 describes combinations and conjugated products of pea protein and polysaccharides, in particular xyloglucans, fucoidans and ulvans, for topical treatment of inflammatory skin disorders such as atopic dermatitis, psoriasis, rosacea and rashes.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1A shows that pre-treatment at various time with formulations 1-3 preserved the reduction in TEER induced by Candida.
  • FIG. 1B shows that pretreatment at various time with formulations 1-3 preserved the membrane integrity of fibroblasts infected with S. aureus.
  • FIG. 2A shows that post-treatment increased TEER 6 and 24 hours after Candida infections.
  • FIG. 2B shows that post-treatment preserved membrane integrity to a time dependent extent as from 6 hours after the S. aureus infection.
  • FIG. 3A shows loss of membrane integrity in fibroblast infected by Candida when the mixtures were administered simultaneously with the infection.
  • FIG. 3B shows limited increased TEER in fibroblast infected by S. aureus when the mixtures were administered simultaneous with the infection.
  • FIG. 4A shows that pre-treatment at various time with formulations 1-3 preserved the membrane properties against Candida infection.
  • FIG. 4B shows that pre-treatment at various time with formulations 1-3 preserved the membrane properties against Gardnerella vaginalis infection.
  • FIG. 5A shows that post-treatment preserved membrane integrity at 6 hours after Candida infection.
  • FIG. 5B shows that post-treatment preserved membrane integrity 24 hours after Candida infection.
  • FIGS. 6A and 6B show that when the mixtures were administered simultaneously with the infection, TEER increased in the infected cells.
    •  DESCRIPTION OF THE INVENTION
  • It has now been found that the combination of pea protein and low-molecular-weight polyphenols belonging to the hydroxybenzoic and hydroxycinnamic acid families is particularly effective in the treatment of skin and mucosal disorders due to impairment of the barrier effect and of the intercellular tight junctions of the dermal and mucosal epithelia.
  • The invention therefore relates to topical formulations comprising combinations or complexes of pea protein with polyphenols belonging to the hydroxycinnamic and hydroxybenzoic acid families.
  • A further aspect of the invention relates to said combinations, complexes and formulations for use in the treatment of skin and mucosal disorders due to impairment of the barrier effect, and in particular in the treatment of local or generalised disorders caused by increased permeation of the natural epithelial barriers.
  • The formulations according to the invention exhibit increased specificity of action and optimum technological characteristics, which allow wide formulation flexibility.
  • The term “skin and mucosal disorders due to impairment of the barrier effect” is clear to the skilled person, as demonstrated, for example, by the following references: Grainne M. O. et al., “Filaggrin in atopic dermatitis”, J. Allergy Clin. Immunol., 2008, vol. 122(4), 689-693; Dorota Purzycka-Bohdan et al., “Genetic background of skin barrier dysfunction in the pathogenesis of psoriasis vulgaris”, Postepy Dermatol Allergol., 2015, 32(2), 123-126; and Flavia Alvim Sant'Anna Addor “Skin barrier in rosacea”, Ann. Bras Dermatol., 2016, vol. 91 (1), 59-63.
  • Atopic dermatitis, psoriasis, rosacea, contact dermatitis, cutaneous candidiasis, fungal or bacterial infections are examples of disorders wherein the barrier effect of the skin is impaired. Vaginal candidiasis is an example of a disorder wherein the barrier effect of the mucosa is impaired.
  • The compositions according to the invention facilitate inhibition of the mechanisms whereby allergens, bacteria, viruses and fungi pass through the epithelia, by reducing the permeability of the tight junctions and inhibiting paracellular transport to other systems. The compositions of the invention also control and reduce water loss, with a consequent reduction in exposure to pro-allergenic factors.
  • The reinstatement of the barrier effect or prevention of its impairment obtainable with the formulations according to the invention also effectively counteract the transfer of the disorder from the epithelium to other systems, a phenomenon known as the “atopic march”, described by Selene K. Bantz et al., “The Atopic March: Progression from Atopic Dermatitis to Allergic Rhinitis and Asthma” in J Clin Cell Immunol. 2014 April; 5(2): 202.
  • “Pea protein” here means a protein powder containing legumin obtained by extraction by known methods from Pisum sativum seeds. Pea protein is available on the market under the trademarks Nutralys®, Pisane® and P80X. Commercially available products can be obtained, for example, from Cosucra and Dal Cin Gildo S.p.a. The product presents as a yellowish powder with a characteristic odour, which is poorly soluble in water.
  • The polyphenols which can be used according to the invention belong to the class of hydroxycinnamic and hydroxybenzoic acids. Examples of said polyphenols comprise ferulic acid, isoferulic acid, chlorogenic acid, ellagic acid, gallic acid and rosmarinic acid, preferably ellagic acid and chlorogenic acid. The polyphenols can be present in a substantially pure, isolated form, or in the form of extracts containing them in percentages ranging from 10 to 80%, depending on the source.
  • A known source of ellagic acid, for example, is pomegranate extract, which has an ellagic acid content typically ranging between 30 and 60% by weight.
  • A known source of chlorogenic acid, for example, is green coffee extract, which has a chlorogenic acid content typically ranging between 35 and 65% by weight.
  • The weight ratio of polyphenol to pea proteins ranges between 0.4:1 and 1:10; the ratio preferably ranges from 1:1 to 1:6. The weight ratio of pea protein to an extract containing 40 to 50% by weight of polyphenol can range from 1:1 to 1:3, preferably 1:1.
  • Examples of suitable forms of administration comprise ointments, lotions, creams, solutions, suspensions, gels, mouthwashes, medicated patches or gauze, powders, pessaries and other forms suitable for topical application to the skin and mucosa.
  • The weight percentage of the pea protein can range from 1 to 30% of the total weight of the formulation, preferably from 1.5 to 10%, while the weight percentage of the polyphenol can range from 0.5 to 25% of the total weight of the formulation.
  • The formulations of the invention are prepared by known methods and with known carriers. The mixture of polyphenol and pea protein according to the invention is preferably associated with moisturising agents, emollients, humectants, preservatives, antioxidants and pH regulators. Examples of said agents are given in WO 2018/167131. Non-lipidic acidifiers such as lectin, lactic acid, sericin, poloxamers, carrageenan and arabinoxylan are particularly preferred. Of these, the following are particularly preferred: one or more of the following compounds selected from lectin, poloxamers, carrageenan and arabinoxylan, each at concentrations ranging from 0.1 to 2.5% by weight.
  • If desired, the formulations according to the invention can contain other active ingredients such as sucralfate, hyaluronic acid, antibiotics, steroidal and non-steroidal anti-inflammatories, chitosan, antihistamines and extracts of medicinal plants, in particular Zanthoxylum bungeanum, Aloe vera and camomile extracts.
  • The compositions according to the invention are particularly useful for the treatment of atopic dermatitis, rosacea, psoriasis, contact dermatitis, dermatitis caused by exposure to sunlight, radiotherapy or chemotherapy, cutaneous candidiasis, fungal infections, vaginal candidiasis, polybacterial vaginitis, fungal and bacterial infections of the skin and mucosa, and infections caused by protozoa.
  • The examples below illustrate the invention in greater detail.
    • Example 1
  • Cream Formulation for Cutaneous Application (Percentages by Weight)
  • Ingredient %
    Pea protein 1.5%  
    Ferulic acid 1.5%  
    Sericin
    2%
    Zanthoxylum b. extract 0.5%  
    Lactic acid 0.3%  
    Palmitate 6%
    Cocoa butter
    1%
    Petrolatum
    1%
    Dimethicone
    2%
    Cetearyl alcohol 3%
    Propylene glycol
    1%
    Glycerin
    1%
    Disodium EDTA 0.2%  
    Phenoxyethanol
    1%
    Dicetylphosphate
    1%
    Water q.s. to 100
    • Example 2
  • Cream Formulation for Cutaneous Application (Percentages by Weight)
  • Ingredient %
    Pea protein 2.5%  
    Chlorogenic acid 2.5%  
    Sericin
    2%
    Zanthoxylum b. extract 0.3%  
    Lectin
    1%
    Palmitate 6%
    Cocoa butter
    1%
    Petrolatum
    1%
    Dimethicone
    2%
    Cetearyl alcohol 3%
    Propylene glycol
    1%
    Glycerin
    1%
    Carrageenan 1%
    Disodium EDTA 0.2%  
    Phenoxyethanol
    1%
    Dicetylphosphate
    1%
    Water q.s. to 100
    • Example 3
  • Cream Formulation for Vaginal Application (Percentages by Weight)
  • Ingredient %
    Pea protein 2.5%  
    Ellagic acid 2.5%  
    Zanthoxylum b. extract 0.2%  
    Arabinoxylan
    2%
    Lactic acid 0.4%  
    Poloxamer
    2%
    Sericin 3%
    Squalane
    2%
    Sodium bicarbonate 3%
    Salcare ® 4%
    Nipaguard ®
    1%
    Water q.s. to 100
    • Example 4
  • Cream Formulation for Vaginal Application (Percentages by Weight)
  • Ingredient %
    Pea protein 2.5%  
    Gallic acid 2.5%  
    Poloxamer
    5%
    Zanthoxylum b. extract 0.5%  
    Lactic acid 0.4%  
    Poloxamer
    2%
    Sericin 3%
    Sodium bicarbonate 3%
    Salcare ® 4%
    Nipaguard ®
    1%
    Carrageenan 1%
    Water q.s. to 100
    • Example 5
  • Three cream formulations containing 5% by weight of the following mixtures:
      • 1. 50% green coffee extract containing 45% chlorogenic acid and 50% pea protein;
      • 2. 40% green coffee extract containing 45% chlorogenic acid and 60% pea protein;
      • 3. 30% green coffee extract containing 45% chlorogenic acid and 70% pea protein,
      • were subjected to the permeability adhesion test (TEER) using murine fibroblasts L-929 which were infected with Staphylococcus aureus and Candida albicans strains.
  • Pre-treatment at various times with formulations 1-3 preserved the reduction in TEER induced by Candida (FIG. 1A) and the membrane integrity of fibroblasts infected with S. aureus (FIG. 1B).
  • Post-treatment increased TEER 6 and 24 hours after the Candida infection (FIG. 2A) and preserved membrane integrity to a time-dependent extent as from 6 hours after the S. aureus infection (FIG. 2B).
  • When the mixtures were administered simultaneously with the infection, loss of membrane integrity (FIG. 3A) in fibroblasts infected by Candida, and increased TEER in fibroblasts infected by S. aureus (FIG. 3B), were again limited.
    • Example 6
  • Three cream formulations containing 5% by weight of the following mixtures:
      • 1. 50% pomegranate extract containing 40% ellagic acid and 50% pea protein;
      • 2. 40% pomegranate extract containing 40% ellagic acid and 60% pea protein;
      • 3. 30% pomegranate extract containing 40% ellagic acid and 70% pea protein,
      • were subjected to the permeability adhesion test (TEER) using a vaginal epithelium cell line VK2 E6/E7 (ATCC CRL-2616) which was infected with Gardnerella vaginalis (GV) and Candida albicans (CA) strains.
  • Pre-treatment at various times with formulations 1-3 preserved the membrane properties against CA and GV infection (FIGS. 4A and 4B).
  • Post-treatment preserved membrane integrity 6 and 24 hours after Candida infection (FIGS. 5A and 5 B).
  • When the mixtures were administered simultaneously with the infection, TEER increased in the infected cells (FIGS. 6A and 6B).

Claims (9)

1. Method of treating localised disorders of bacterial, viral, inflammatory, allergic and/or fungal, endogenous, or idiopathic origin caused by permeation of the natural epithelial barriers, and disorders transferred from the epithelia to other systems in a subject in need thereof, said method comprising:
administering a pharmaceutically effective amount of topical the compositions comprising pea protein and polyphenols of the hydroxycinnamic and hydroxybenzoic acid families selected from ferulic acid, isoferulic acid, gallic acid and rosmarinic acid wherein the weight ratio of polyphenol to pea protein is from 0.4:1 to 1:10, in admixture with a suitable carrier to said subject.
2. The method according to claim 1, wherein said localized disorders comprise atopic dermatitis, rosacea, psoriasis, contact dermatitis, dermatitis caused by exposure to sunlight, radiotherapy or chemotherapy, cutaneous candidiasis, fungal infections, vaginal candidiasis, polybacterial vaginitis, fungal and bacterial infections of the skin and mucous membranes, and infections caused by protozoa.
3. The method according to claim 1, wherein the polyphenol is ferulic acid.
4. The method according to claim 1, wherein the weight ratio of polyphenol to pea protein ranges from 1:1 to 1:6.
5. The method according to claim 1, wherein the weight ratio of polyphenol to pea protein is 1:1.
6. The method according to claim 1, wherein the compositions are in the form of ointments, lotions, creams, solutions, suspensions, gels, mouthwashes, medicated plasters or gauzes, powders, or pessaries.
7. The method according to claim 1, wherein the compositions further comprise moisturising, emollient, humectant, preservative, antioxidant, or pH-regulating additives.
8. The method according to claim 7, wherein the additives are selected from lectin, lactic acid, sericin, poloxamers, carrageenan and arabinoxylan.
9. The method according to claim 8, wherein the additives are selected from lectin, poloxamers, carrageenan and arabinoxylan, each in concentrations by weight ranging from 0.1 to 2.5%.
US18/464,295 2019-12-23 2023-09-11 Topical compositions comprising pea proteins and polyphenols Abandoned US20230414708A1 (en)

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