WO2025080692A1 - Compositions and methods for repelling insects and arachnids - Google Patents

Abstract

Provided herein are composition and methods for repelling an arachnid or an insect from a subject or a fabric comprising applying a composition comprising one or more of hemoglobin, hemocyanin and myoglobin. Also provided are compositions comprising two or more of hemoglobin, hemocyanin and myoglobin.

Description

^ COMPOSITIONS AND METHODS FOR REPELLING INSECTS AND ARACHNIDS ^ CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to, and the benefit of, U.S. Provisional Patent Application No. 63/588,801, filed October 9, 2023, which is incorporated by reference herein in its entirety. REFERNCE TO SEQUENCE LISTING [0002] The sequence listing submitted on October 9, 2024, as an .XML entitled “10504- 097WO1_ST26.xml” created on October 9, 2024, and having a file size of 13,244 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.52(e)(5). BACKGROUND OF THE INVENTION [0003] Ixodes scapularis, or the black-legged tick, is a vector for human diseases including but not limited to Lyme disease in the northeastern United States. In the last century, the northeastern United States, especially Pennsylvania, has become very favorable for the ecology of tick-borne diseases. The region’s woodlands, rivers, growing Odocoileus virginianus or white-tailed deer population, plentiful population of the Peromyscus leucopusor or white footed mouse, and the expansion of human populations into the black-legged tick’s habitat yield the ideal conditions for the transmission of Lyme disease and the other six pathogens to humans. [0004] Black-legged ticks can become infected with and transmit up to seven different pathogens during their life cycle, which spans 2 years. First, eggs hatch into larva, which will molt into nymphs. Nymphs then grow into adults, where the females lay eggs, perpetuating the cycle. I. scapularis must feed, or take one blood meal, during each stage of life and may acquire these pathogens during their larval or nymph stage when feeding on the reservoirs, such as P. leucopusor. During the next stage of the life cycle, the tick may feed on O. virginianus, its definitive host, or an accidental host, such as humans or domesticated animals where transmission can occur. Most infections occur between early spring and late summer through nymphs, because they are much smaller and harder to detect than adults.

^^ ^ ^ [0005] Black-legged ticks are active host seekers that are intensely attracted to carbon dioxide (CO2). A study showed that humans, under laboratory conditions, produced CO2 at the rate of 1.0-1.8 ml/hour on the hand. Increased production of CO2 was observed with an increase in temperature, as to be expected. Another study looked at carbon dioxide emission from human skin, in a more natural setting, determined that the emission rate was 3.4 x 10-5 ml CO₂/cm/minute for the arm and forearm combined. Emission rates are increased to 8.6 x 10-5 ml CO2/cm/minute after vigorous exercise, and 9.1 x 10-5 ml CO2/cm/minute after wetting the skin. [0006] Many studies have sought to determine if natural compounds derived from plants can act as repellents for these arachnids with varying levels of success. Interest is high in natural repellents for blood-sucking vectors because of the wide public concern regarding DEET. DEET has been correlated to seizures and encephalopathy in children. [0007] As a result of these concerns, what are needed are improved repellents of arachnids and insects. Also needed are methods for increasing a skin surface oxygenation level. SUMMARY OF THE INVENTION [0008] Provided herein is a method of repelling an arachnid or an insect from a subject or a fabric comprising applying a composition to the subject or the fabric, wherein the composition comprises one or more of hemoglobin, hemocyanin and myoglobin, and wherein the application repels the arachnid or the insect as compared to a control. [0009] In some embodiments, the composition comprises hemoglobin and hemocyanin. In some embodiments, the composition comprises hemocyanin and myoglobin. In some embodiments, the composition comprises myoglobin and hemoglobin. In some embodiments, the composition comprises hemoglobin. [0010] In some aspects, the hemoglobin comprises one or more of an ^1, ^2, ^1 and ^2 subunit. In some embodiments, the hemoglobin comprises one or more of SEQ ID NO: 1 and SEQ ID NO:2, or a sequence having at least 80% identity with SEQ ID NO: 1 and SEQ ID NO:2, or a fragment SEQ ID NO: 1 or SEQ ID NO:2. [0011] In some aspects, the composition is applied to the subject’s skin. In other aspects, the composition is applied to the fabric. [0012] In some embodiments, the arachnid is a tick. In some embodiments, the tick is a black legged tick.

^^ ^ ^ [0013] In some aspects, the subject is a human. [0014] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier and/or solvent. The pharmaceutically acceptable carrier can comprise glycerol, water, petroleum jelly, aloe vera, coconut oil, and/or avocado oil. [0015] In some embodiments, the arachnid or insect is repelled in an amount of at least 50% as compared to a control. In some embodiments, the arachnid or insect is repelled in an amount of at least 75% as compared to a control. In some embodiments, the arachnid or insect is repelled in an amount of at least 90% as compared to a control. [0016] Also included herein is a composition comprising two or more of hemoglobin, hemocyanin and myoglobin. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can comprise glycerol, water, petroleum jelly, aloe vera, coconut oil, and/or avocado oil. [0017] In some embodiments, the composition comprises hemoglobin and hemocyanin. In some embodiments, the composition comprises hemocyanin and myoglobin. In some embodiments, the composition comprises myoglobin and hemoglobin. BRIEF DESCRIPTION OF FIGURES [0018] Figure 1(A-B) shows that BITE about doubles latency compared to the vehicle alone and more than quadruples latency compared to no treatment. Fig.1(A) is a graph showing that BITE significantly delayed tick chemotaxis as compared to vehicle alone and no treatment (latency denoted in seconds; Wilcoxon Signed Rank: Z=-2.328, p=0.020). Figure 1(B) provides descriptive statistics. DETAILED DESCRIPTION [0019] Provided herein are compositions and methods of repelling an arachnid or insect from a subject or a fabric that includes applying a composition to the subject or the fabric, wherein the composition comprises one or more of hemoglobin, hemocyanin and myoglobin. Also provided herein are compositions and methods of increasing a skin oxygenation level comprising applying a composition to the skin, wherein the composition comprises one or more of hemoglobin, hemocyanin and myoglobin. [0020] The following description of the disclosure is provided as an enabling teaching of the disclosure in its best, currently known embodiment(s). To this end, those skilled in the relevant art will recognize and appreciate that many changes can be made to the various

^^ ^ ^ embodiments of the invention described herein, while still obtaining the beneficial results of the present disclosure. It will also be apparent that some of the desired benefits of the present disclosure can be obtained by selecting some of the features of the present disclosure without utilizing other features. Accordingly, those who work in the art will recognize that many modifications and adaptations to the present disclosure are possible and can even be desirable in certain circumstances and are a part of the present disclosure. Thus, the following description is provided as illustrative of the principles of the present disclosure and not in limitation thereof. [0021] Reference will now be made in detail to the embodiments of the invention, examples of which are illustrated in the drawings and the examples. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. [0022] Terminology [0023] The following definitions are provided for the full understanding of terms used in this specification. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. [0024] As used in this disclosure and in the appended claims, the singular forms “a”, “an”, “the”, include plural referents unless the context clearly dictates otherwise. For example, the term "a cell" includes a plurality of cells, including mixtures thereof. [0025] The terms "about" and "approximately" are defined as being “close to” as understood by one of ordinary skill in the art. In one non-limiting embodiment the terms are defined to be within 10%. In another non-limiting embodiment, the terms are defined to be within 5%. In still another non-limiting embodiment, the terms are defined to be within 1%. [0026] As used herein, “apply,” “applying,” or “applied” means to place onto or into. These terms include, but are not limited to, spraying, spreading and embedding. A surface having such an application is also referred to herein as a “treated surface.” [0027] A "composition" is intended to include a combination of active agent and another compound or composition, inert (for example, a detectable agent or label) or active, such as an adjuvant. [0028] The term “comprising”, and variations thereof as used herein is used synonymously with the term “including” and variations thereof and are open, non-limiting terms. Although

^^ ^ ^ the terms “comprising” and “including” have been used herein to describe various embodiments, the terms “consisting essentially of” and “consisting of” can be used in place of “comprising” and “including” to provide for more specific embodiments and are also disclosed. [0029] A "control" is an alternative subject or sample used in an experiment for comparison purposes. A control can be "positive" or "negative." [0030] The term “pharmaceutically acceptable carrier” means a carrier or excipient that is useful in preparing a pharmaceutical composition that is generally safe and non-toxic and includes a carrier that is acceptable for veterinary and/or human pharmaceutical or topical use. As used herein, the term "pharmaceutically acceptable carrier" encompasses any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, and emulsions, such as an oil/water or water/oil emulsion, and various types of wetting agents. As used herein, the term “carrier” encompasses any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other material well known in the art for use in pharmaceutical formulations. The compositions described herein also can include preservatives. A “pharmaceutically acceptable carrier” as used in the specification and claims includes both one and more than one such carrier. [0031] As used herein, “reduce” means to decrease by a statistically significant amount. In some embodiments, the reduction is about 10%, 20%, 30%, 40%, 50%, 60%, 70, 80%, or 90%. [0032] As used herein, “repel,” “repelling” or any grammatical form thereof, refers to a reduction in the movement of an arachnid or insect toward or onto a treated surface. [0033] The term “subject” is defined herein to include animals such as mammals, including, but not limited to, primates (e.g., humans), pets, livestock animals, beasts of burden, and rodents. The term “pets” includes, but is not limited to, cats and dogs. The term “livestock animals” includes, but is not limited to cows, sheep, goats, and pigs. The term “beasts of burden” includes, but is not limited to, horses, camels, and donkeys. The term “rodents” includes, but is not limited to, mice, rats, squirrels, prairie dogs, guinea pigs, and hamsters. In some embodiments, the subject is a human. In some embodiments, the subject is a pet. [0034] Compositions and Methods [0035] Provided herein is a method of repelling an arachnid or insect from a subject or a fabric comprising applying a composition to the subject or fabric, wherein the composition

^^ ^ ^ comprises one or more of hemoglobin, hemocyanin and myoglobin. Also provided herein are compositions comprising one or more of hemoglobin, hemocyanin and myoglobin. [0036] In some embodiments, the composition comprises hemoglobin. In some embodiments, the composition comprises hemoglobin and not hemocyanin or myoglobin. In some embodiments, the composition comprises hemocyanin and not hemoglobin or myoglobin. In some embodiments, the composition comprises myoglobin and not hemoglobin or hemocyanin. In some aspects, the composition comprises hemoglobin and hemocyanin. In other aspects, the composition comprises hemoglobin and myoglobin. In still other aspects, the composition comprises hemocyanin, myoglobin and hemocyanin. [0037] As used herein “hemoglobin” refers to an oxygen-, carbon dioxide- and iron-binding protein that can be found in blood or hemolymph. In some aspects, the hemoglobin is human hemoglobin. In some embodiments, the hemoglobin is a tetramer having two alpha subunits (^1 and ^2) and two beta subunits (^1 and ^2), wherein the alpha and beta subunits form two ^^ dimers (^1^1 and ^2^2). In some embodiments, the hemoglobin comprises one, two or three subunits of human hemoglobin, for example, ^1, ^2, and ^1, or ^1, ^2, ^2, or ^1, ^1 and ^2, or ^2, ^1 and ^2. In some embodiments, the hemoglobin is a hemoglobin A or adult hemoglobin. In some embodiments, the alpha subunit comprises SEQ ID NO: 2. In other embodiments, the hemoglobin alpha subunit described herein comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2. In some embodiments, the beta subunit comprises SEQ ID NO: 1. In other embodiments, the hemoglobin beta subunit described herein comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 1 or a fragment of SEQ ID NO: 1. [0038] In some aspects, the hemoglobin is a crustacean hemoglobin. In some embodiments, the hemoglobin comprises SEQ ID NO: 3. In other embodiments, the hemoglobin comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 3 or a fragment of SEQ ID NO:3. In some embodiments, the hemoglobin comprises SEQ ID NO: 8. In other embodiments, the hemoglobin comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 8 or a fragment of SEQ ID NO: 8.

^^ ^ ^ [0039] In some aspects, the hemoglobin is a Daphnia magna hemoglobin. In some embodiments, the hemoglobin comprises one or more of SEQ ID NOs: 4-7. In other embodiments, the hemoglobin comprises polypeptide sequences at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to one or more of SEQ ID NOs: 4-7 or a fragment of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7. Exemplary but non-limiting fragments of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 include sequences that exclude the signal peptide and/or N-terminal extension. [0040] “Hemocyanin” refers to an oxygen- and carbon dioxide-binding protein that can be found in arthropod or mollusk species. Hemocyanins comprise monomers of approximately 75 kDa that form one or more dimers or hexamers. When there is more than one dimer or hexamer, the dimers or hexamers form complexes. For example, hemocyanin from spiny lobster has one hexamer, hemocyanin from lobster and crab has two hexamers, hemocyanin from scorpion and spider has four hexamers, hemocyanin from centipede has six hexamers, hemocyanin from horseshoe crab has eight hexamers, hemocyanin from octopus, cuttle fish, squid and chiton has five dimers, and hemocyanin from snail has ten dimers. Accordingly, in some aspects, the hemocyanin is a spiny lobster hemocyanin, a lobster hemocyanin, a crab hemocyanin, a scorpion hemocyanin, a spider hemocyanin, a centipede hemocyanin, or a horseshoe crab hemocyanin. In some embodiments, the hemocyanin is an octopus hemocyanin, a cuttle fish hemocyanin, a squid hemocyanin, a chiton hemocyanin, or a snail hemocyanin. In some embodiments, the hemocyanin comprises SEQ ID NO: 9. In other embodiments, the hemocyanin comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 9 or a fragment of SEQ ID NO: 9. In some embodiments, the hemocyanin comprises SEQ ID NO: 10. In other embodiments, the hemocyanin comprises a polypeptide sequence at least about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 10 or a fragment of SEQ ID NO: 10. [0041] As used herein, the term “myoglobin” refers to an oxygen- and carbon dioxide-binding protein that can be found in muscles. In some aspects, the myoglobin is human. In some embodiments, the myoglobin comprises SEQ ID NO: 11. In other embodiments, the myoglobin comprises a polypeptide sequence at least about 80%, about 85%, about 90%,

^^ ^ ^ about 95%, about 97%, about 98%, about 99%, or about 99.5% identical to SEQ ID NO: 11 or a fragment of SEQ ID NO: 11. [0042] In certain aspects, the hemoglobin, hemocyanin and/or myoglobin is about 1% to about 30% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is about 5% to about 15% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is about 8% to about 12% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is about 10% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% by weight of the composition. In certain aspects, the hemoglobin, hemocyanin and/or myoglobin is at least about 1% to at least about 20% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is at least about 5% to at least about 15% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is at least about 8% to at least about 12% by weight of the composition. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin is at least about 10% by weight of the composition. [0043] The hemoglobin, hemocyanin and/or myoglobin in the composition can also be dissolved in a pharmaceutically acceptable solvent. Suitable solvents include, but are not limited to, polyethylene glycol (PEG), diglycol monoethyl ether; alklene glycols, such as propylene glycol; dimethyl isosorbide; alcohols, such as isopropyl alcohol and ethanol. The solvents are typically selected for their ability to dissolve the drug. In some embodiments, the solvent for the hemoglobin, hemocyanin and/or myoglobin comprises polyethylene glycol (PEG). In some embodiments, the solvent for the hemoglobin, hemocyanin and/or myoglobin comprises PEG 400. [0044] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can comprise glycerol, water, petroleum jelly, aloe vera, coconut oil, and/or avocado oil. In some embodiments, the pharmaceutically acceptable carrier comprises aloe vera. In some embodiments, the pharmaceutically acceptable carrier comprises water. In some embodiments, the pharmaceutically acceptable carrier comprises aloe vera and water. In some embodiments, the

^^ ^ ^ aloe vera is about 20% to about 80% by weight of the composition or the pharmaceutically acceptable carrier. In some embodiments, the aloe vera is about 30% to about 70% by weight of the composition or the pharmaceutically acceptable carrier. In some embodiments, the aloe vera is about 40% to about 60% by weight of the composition or the pharmaceutically acceptable carrier. In some embodiments, the aloe vera is about 45% to about 55% by weight of the composition or the pharmaceutically acceptable carrier. In some embodiments, the aloe vera is about 50% by weight of the composition or the pharmaceutically acceptable carrier. [0045] The arachnid or insect repelled using the methods described herein can be any. In some embodiments, the arachnid is a tick. The tick can be any tick. In some embodiments the tick can be selected from the group consisting of blacklegged tick (Ixodes scapularis), lone star tick (Amblyomma americanum), american dog tick (Dermacentor variabilis, d. Similis), brown dog tick (Rhipicephalus sanguineus), groundhog tick (Ixodes cookie), gulf coast tick (Amblyomma maculatum), rocky mountain wood tick (Dermacentor andersoni), soft tick (Ornithodoros spp.), and western blacklegged tick (Ixodes pacificus). In some embodiments, the tick is a black legged tick. In some embodiments, the insect is a mosquito. [0046] The application of the composition to the subject or fabric can be via any means. Included herein are applications made by spreading and spraying. Also included herein are applications via embedding the composition into the fabric. In some embodiments, the composition is applied topically to the subject. In some embodiments, the composition is applied to the subject’s exposed skin. In some embodiments, the composition is applied to the subject’s clothing. [0047] The composition can be applied in such amounts, time, and route deemed necessary in order to achieve the desired result. The exact amount of the composition will vary from subject to subject, depending on the species, age, and general condition of the subject, the particular compound, its mode of application, its mode of activity, and the like. The specific effective application amount for any particular subject will depend upon a variety of factors including the activity of the composition employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration and the amount of carbon dioxide produced by the subject; the number of applications; and like factors well known in the medical arts. [0048] Topical Compositions

^^ ^ ^ [0049] Dosage forms for topical administration of the hemoglobin, hemocyanin and/or myoglobin compounds described herein include ointments, gels, pastes, liquids, solutions, creams, oils, powders, aerosols, and sprays. Thickening agents, emollients, and stabilizers can be used to prepare the topical compositions of the present invention. Examples of thickening agents include petrolatum, beeswax, xantham gum, or polyethylene glycol, humectants such as sorbitol, emollients such as mineral oil, lanolin and its derivatives, or squalene. The hemoglobin, hemocyanin and/or myoglobin compounds described herein are admixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, thickening agents, cosmetic agents, or propellants as can be required. [0050] The therapeutically effective amount of the topical hemoglobin, hemocyanin and/or myoglobin compositions described herein can be determined by one of ordinary skill in the art and includes exemplary dosage amounts for a mammal of from about 0.5 to about 200 mg/kg of body weight of active composition per day, which can be administered in a single dose or in the form of individual divided doses, such as from 1 to 4 times per day. Alternatively, the dosage amount can be from about 0.5 to about 150 mg/kg of body weight of active composition per day, about 0.5 to 100 mg/kg of body weight of active compound per day, about 0.5 to about 75 mg/kg of body weight of active compound per day, about 0.5 to about 50 mg/kg of body weight of active composition per day, about 0.5 to about 25 mg/kg of body weight of active composition per day, about 1 to about 20 mg/kg of body weight of active composition per day, about 1 to about 10 mg/kg of body weight of active composition per day, about 20 mg/kg of body weight of active composition per day, about 10 mg/kg of body weight of active composition per day, or about 5 mg/kg of body weight of active composition per day. [0051] Those of skill in the art will understand that the specific dose level and frequency of dosage for any particular subject can be varied and will depend upon a variety of factors, including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the species, age, body weight, general health, sex and diet of the subject, the mode and time of administration, rate of excretion, drug combination, and severity of the particular condition. [0052] An “emulsion” is a preparation of one liquid distributed in small globules throughout the body of a second liquid. The dispersed liquid is the discontinuous phase, and the dispersion medium is the continuous phase. When oil is the dispersed liquid and an aqueous solution is the

^^^ ^ ^ continuous phase, it is known as an oil-in-water emulsion, whereas when water or aqueous solution is the dispersed phase and oil or oleaginous substance is the continuous phase, it is known as a water-in-oil emulsion. The oil phase may consist at least in part of a propellant, such as an HFA propellant. Either or both of the oil phase and the aqueous phase may contain one or more surfactants, emulsifiers, emulsion stabilizers, buffers, and other excipients. Preferred excipients include surfactants, especially non-ionic surfactants; emulsifying agents, especially emulsifying waxes; and liquid non-volatile non-aqueous materials, particularly glycols such as propylene glycol. The oil phase may contain other oily pharmaceutically approved excipients. For example, materials such as hydroxylated castor oil or sesame oil may be used in the oil phase as surfactants or emulsifiers. [0053] "Emollients" are an externally applied agent that softens or soothes skin and are generally known in the art. These include, without limitation, almond oil, castor oil, ceratonia extract, cetostearoyl alcohol, cetyl alcohol, cetyl esters wax, cholesterol, cottonseed oil, cyclomethicone, ethylene glycol palmitostearate, glycerin, glycerin monostearate, glyceryl monooleate, isopropyl myristate, isopropyl palmitate, lanolin, lecithin, light mineral oil, medium-chain triglycerides, mineral oil and lanolin alcohols, petrolatum, petrolatum and lanolin alcohols, soybean oil, starch, stearyl alcohol, sunflower oil, xylitol and combinations thereof. In one embodiment, the emollients are ethylhexylstearate and ethylhexyl palmitate. [0054] "Surfactants" are surface-active agents that lower surface tension and thereby increase the emulsifying, foaming, dispersing, spreading and wetting properties of a product. Suitable non-ionic surfactants include emulsifying wax, glyceryl monooleate, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polysorbate, sorbitan esters, benzyl alcohol, benzyl benzoate, cyclodextrins, glycerin monostearate, poloxamer, povidone and combinations thereof. [0055] "Emulsifiers" are surface active substances which promote the suspension of one liquid in another and promote the formation of a stable mixture, or emulsion, of oil and water. Common emulsifiers are: metallic soaps, certain animal and vegetable oils, and various polar compounds. Suitable emulsifiers include acacia, anionic emulsifying wax, calcium stearate, carbomers, cetostearyl alcohol, cetyl alcohol, cholesterol, diethanolamine, ethylene glycol palmitostearate, glycerin monostearate, glyceryl monooleate, hydroxpropyl cellulose, hypromellose, lanolin, hydrous, lanolin alcohols, lecithin, medium-chain triglycerides,

^^^ ^ ^ methylcellulose, mineral oil and lanolin alcohols, monobasic sodium phosphate, monoethanolamine, nonionic emulsifying wax, oleic acid, poloxamer, poloxamers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, propylene glycol alginate, self-emulsifying glyceryl monostearate, sodium citrate dehydrate, sodium lauryl sulfate, sorbitan esters, stearic acid, sunflower oil, tragacanth, triethanolamine, xanthan gum and combinations thereof. [0056] A "lotion" is a low- to medium-viscosity liquid formulation. A lotion can contain finely powdered substances that are in soluble in the dispersion medium through the use of suspending agents and dispersing agents. Alternatively, lotions can have as the dispersed phase liquid substances that are immiscible with the vehicle and are usually dispersed by means of emulsifying agents or other suitable stabilizers. In one embodiment, the lotion is in the form of an emulsion having a viscosity of between 100 and 1000 centistokes. The fluidity of lotions permits rapid and uniform application over a wide surface area. Lotions are typically intended to dry on the skin leaving a thin coat of their medicinal components on the skin's surface. [0057] A "cream" is a viscous liquid or semi-solid emulsion of either the "oil-in-water" or "water-in-oil type". Creams may contain emulsifying agents and/or other stabilizing agents. In one embodiment, the formulation is in the form of a cream having a viscosity of greater than 1000 centistokes, typically in the range of 20,000-50,000 centistokes. Creams are often time preferred over ointments as they are generally easier to spread and easier to remove. [0058] The basic difference between a cream and a lotion is the viscosity, which is dependent on the amount/use of various oils and the percentage of water used to prepare the formulations. Creams are typically thicker than lotions, may have various uses and often one uses more varied oils/butters, depending upon the desired effect upon the skin. In a cream formulation, the water- base percentage is about 60-75% and the oil-base is about 20-30% of the total, with the other percentages being the emulsifier agent, preservatives and additives for a total of 100%. [0059] An "ointment" is a semisolid preparation containing an ointment base and optionally one or more active agents. Examples of suitable ointment bases include hydrocarbon bases (e.g., petrolatum, white petrolatum, yellow ointment, and mineral oil); absorption bases (hydrophilic petrolatum, anhydrous lanolin, lanolin, and cold cream); water-removable bases (e.g., hydrophilic ointment), and water-soluble bases (e.g., polyethylene glycol ointments). Pastes

^^^ ^ ^ typically differ from ointments in that they contain a larger percentage of solids. Pastes are typically more absorptive and less greasy that ointments prepared with the same components. [0060] A "gel" is a semisolid system containing dispersions of small or large molecules in a liquid vehicle that is rendered semisolid by the action of a thickening agent or polymeric material dissolved or suspended in the liquid vehicle. The liquid may include a lipophilic component, an aqueous component or both. Some emulsions may be gels or otherwise include a gel component. Some gels, however, are not emulsions because they do not contain a homogenized blend of immiscible components. Suitable gelling agents include, but are not limited to, modified celluloses, such as hydroxypropyl cellulose and hydroxyethyl cellulose; Carbopol homopolymers and copolymers; and combinations thereof. Suitable solvents in the liquid vehicle include, but are not limited to, diglycol monoethyl ether; alklene glycols, such as propylene glycol; dimethyl isosorbide; alcohols, such as isopropyl alcohol and ethanol. The solvents are typically selected for their ability to dissolve the drug. Other additives, which improve the skin feel and/or emolliency of the formulation, may also be incorporated. Examples of such additives include, but are not limited, isopropyl myristate, ethyl acetate, C12-C15 alkyl benzoates, mineral oil, squalane, cyclomethicone, capric/caprylic triglycerides, and combinations thereof. [0061] Foams consist of an emulsion in combination with a gaseous propellant. The gaseous propellant consists primarily of hydrofluoroalkanes (HFAs). Suitable propellants include HFAs such as 1,1,1,2-tetrafluoroethane (HFA 134a) and 1,1,1,2,3,3,3-heptafluoropropane (HFA 227), but mixtures and admixtures of these and other HFAs that are currently approved or may become approved for medical use are suitable. The propellants preferably are not hydrocarbon propellant gases which can produce flammable or explosive vapors during spraying. Furthermore, the compositions preferably contain no volatile alcohols, which can produce flammable or explosive vapors during use. [0062] Buffers are used to control pH of a composition. Preferably, the buffers buffer the composition from a pH of about 4 to a pH of about 7.5, more preferably from a pH of about 4 to a pH of about 7, and most preferably from a pH of about 5 to a pH of about 7. In a preferred embodiment, the buffer is triethanolamine. [0063] Preservatives can be used to prevent the growth of fungi and microorganisms. Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid, butylparaben,

^^^ ^ ^ ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, and thimerosal. [0064] Penetration enhancers are frequently used to promote transdermal delivery of drugs across the skin. Certain penetration enhancers can cause some dermal irritation, dermal toxicity and dermal allergies. However, the more commonly used penetration enhancers include urea, (carbonyldiamide), imidurea, N,N-diethylformamide, N-methyl-2-pyrrolidine, 1-dodecal- azacyclopheptane-2-one, calcium thioglycate, 2-pyyrolidine, N,N-diethyl-m-toluamide, oleic acid and its ester derivatives, such as methyl, ethyl, propyl, isopropyl, butyl, vinyl and glycerylmonooleate, sorbitan esters, such as sorbitan monolaurate and sorbitan monooleate, other fatty acid esters such as isopropyl laurate, isopropyl myristate, isopropyl palmitate, diisopropyl adipate, propylene glycol monolaurate, propylene glycol monooleatea and non- ionic detergents such as BRIJ.RTM. 76 (stearyl poly(10 oxyethylene ether), BRIJ.RTM. 78 (stearyl poly(20)oxyethylene ether), BRIJ.RTM. 96 (oleyl poly(10)oxyethylene ether), and BRIJ.RTM.721 (stearyl poly(21)oxyethylene ether) (ICI Americas Inc. Corp.). [0065] In some embodiments, the hemoglobin, hemocyanin and/or myoglobin composition is administered about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours or about 6 hours prior to the subject’s or the fabric’s exposure to the arachnid or insect. In some embodiments, the hemoglobin, hemocyanin and/or myoglobin composition is administered to the fabric a month or more, two months or more, three months or more, four months or more, five months or more or six months or more prior to the fabric’s exposure to the arachnid or insect. [0066] In some embodiments, the subject is a human. In other embodiments the subject is a dog or cat. [0067] In some aspects, the method described herein repels the arachnid or insect in an amount of about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, or more as compared to a control. [0068] Also provided herein is a method of increasing a skin surface oxygenation level comprising applying a composition to the skin surface, wherein the composition comprises one or more of hemoglobin, hemocyanin and myoglobin. The composition can be any as

^^^ ^ ^ described herein. The term “skin surface oxygenation level” means a level of oxygen sequestration on the skin surface. In some embodiments, the skin surface oxygenation level is increased by an amount of about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, or more as compared to a control. [0069] The following examples are set forth below to illustrate the compositions, devices, methods, and results according to the disclosed subject matter. These examples are not intended to be inclusive of all aspects of the subject matter disclosed herein, but rather to illustrate representative methods and results. These examples are not intended to exclude equivalents and variations of the present invention which are apparent to one skilled in the art. [0070] EXAMPLES [0071] Example 1 [0072] Method: The binding affinity of hemoglobin to CO₂ was determined at atmospheric levels of CO2. The binding affinity was monitored by qualitative ultraviolet-visible (UV-Vis) spectroscopy. Carbon dioxide exhibited a characteristic absorption band in the UV-Vis spectrum around 580 nm in deionized water and 980 nm in 20% polyethylene glycol-400 (PEG-400). Samples of hemoglobin were dissolved in deionized water and 20% aqueous PEG-400 and a background IR spectrum was collected. [0073] The samples were then saturated with CO₂ via bubbling gas through the solvent until no spectroscopic change was observed (20 minutes). Reversibility of the binding between CO₂ and each of hemoglobin under atmospheric conditions was established by allowing the sample to equilibrate at room temperature monitored over time. By collecting the background of the compound alone, the CO2 band could be isolated for qualitative study. [0074] Next, the hemoglobin dissolved in deionized water and 20% aqueous PEG-400 was tested for the ability to repel questing ticks on an apparatus designed to mimic CO2 exuded from human skin (“tick arena”) using a version of the tick carousel assay. The tick arena was temperature-monitored and had an inlet through which CO₂ was allowed to enter. The CO₂ inlet was covered with filter paper coated in formulations of aloe/water, either prepared without special considerations or placed under sonication for 30 minutes while being flushed with nitrogen as well as aloe/water solutions containing hemoglobin that were also placed under sonication while being flushed with nitrogen gas for 30 minutes.

^^^ ^ ^ [0075] Example 2 [0076] Forty female ticks were tested using a 10% by weight hemoglobin composition in the tick carousel assay described in Example 1. Specifically, a hemoglobin composition of 10% hemoglobin by weight in 50% aloe/water (referred to herein as “BITE”) was applied to the surface of the inlet filter of the tick arena. Hemoglobin from bovine blood was purchased from Sigma-Aldrich (Catalog: H2625-25G) and used as is. Aloe was purchased from Wal- Mart (Fruit of the Earth) and used as is. Water was purified by reverse osmosis filtration. Solutions were prepared and degassed under sonication for 30 minutes while being flushed with nitrogen gas. The treated filters were placed in front of the CO2 inlet at 70 – 80 °F and tick latency, or delay in tick chemotaxis, was observed. CO2 emission from a controlled regulator was set at 0.5 sccm (equivalent to 1 cm³ STP/min, Nml/min). Figure 1(A) is a boxplot showing that BITE significantly delayed tick chemotaxis as compared to vehicle alone and no treatment (latency denoted in seconds; Wilcoxon Signed Rank: Z=-2.328, p=0.020). Figure 1(B) provides descriptive statistics. Medians and quartiles are used because there is an artificial maximum so the data is not normally distributed. Accordingly, Figure 1(A-B) demonstrates that BITE about doubles latency compared to the vehicle alone and more than quadruples latency compared to no treatment. [0077] It should be understood that the foregoing relates to preferred embodiments of the present invention and that numerous changes may be made therein without departing from the scope of the invention. The invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof. On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof, which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the present invention and/or the scope of the appended claims. All patents, patent applications, and publications referenced herein are incorporated by reference in their entirety for all purposes. [0078] SEQUENCES [0079] SEQ ID NO: 1 Human Hemoglobin beta subunit [0080] MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLST PDAVMGNPKVKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLL GNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVANALAHKYH

^^^ ^ ^ [0081] SEQ ID NO: 2 Human Hemoglobin Alpha subunit [0082] MVLSPADKTNVKAAWGKVGAHAGEYGAEALERMFLSFPTTKTYFPHFDLSH GSAQVKGHGKKVADALTNAVAHVDDMPNALSALSDLHAHKLRVDPVNFKLLSHCL LVTLAAHLPAEFTPAVHASLDKFLASVSTVLTSKYR [0083] SEQ ID NO: 3 Crustacean Hemoglobin [0084] Artemia E1 Domain hemoglobin [0085] ERVDPITGLSGLEKNAILDTWGKVRGNLQEVGKATFGKLFAAHPEYQQMFRF FQGVQLAFLVQSPKFAAHTQRVVSALDQTLLALNRPSDQFVYMIKELGLDHINRGTD RSFVEYLKESLGDSVDEFTVQSFGEVIVNFLNEGLRQA [0086] SEQ ID NOS: 4 Daphnia magna Hemoglobin chain 2 [0087] MQVLTIALFLGIVATASCASMAAPGTTVTTVTTSVTTVSADEESTGILSANERS LIRKTWDQAKKDGDVAPQVLFRFIKAHPEYQKKFSKFADVPQSELLTNGNFLAQAYT ILAGLNVVIQSLSSQELLANQLNVLGGAHQPRGVTPAMFEEFGVIVEQVLEEELGNGF SAEARQAMKHGIHALVAGVSKTLKNPEDLADPQTKLTLHQIRDVQRSMENIRSGRN ALVSSIFVKLFKETPRVQKHFAKFSSVAVDSLAGHADYEKQVALVADRLDTIISAMD DKLQLLGNIYYMKYTHLERGISRDTFEDFGRLLLDVLGASGVSSDDLDSWKGVLAVF VNGVSPKK [0088] SEQ ID NOS: 5 Daphnia magna Hemoglobin chain 1 [0089] MASFKIALLLGVIAFVNACSQAPGTTTTTVTTTVTTVSADDGSEAGLLSAHER SLIRKTWDQAKKDGDVAPQVLFRFVKAHPEYQKMFSKFANVPQSELLSNGNFLAQA YTILAGLNVVIQSLFSQELMANQLNALGGAHQPRGATPVMFEQFGGILEEVLAEELGS GFTAEARQAMKNGLAALVAGIAKNLKKAEDLADPQTKLTPHQIRDVQRSMENIRND RNALVSSIFVKLFKETPRIQKFFAKFANVAVDSLAGNAEYEKQIALVADRLDTMISAM DDKLQLLGNINYMRYTHTERGIPRAPWEDFSRLLLDVLGSKGVSTDDLDSWKGVLA VFVNGVSPIKK [0090] SEQ ID NOS: 6 Daphnia magna Hemoglobin chain 3 [0091] MASFKIALLFGVVAFVSAYSQAPGTTTTTVTTTVTTVSADDGSEAGILSAHER SLIRKTWEQARKDGDVAPQILFRFVKAHPEYQKMFSKFANVPQAELLTNGNFLAQA YTILAGLNVVVQSLSSQELMANQLNALGGAHKPRGATPVMFEQFGGILEEVLAEELG SSFTAEARQAMKNGLTALISGISKNLKKAEDLADPQTKLTPHQIQDVQRSMENIRNGR NALVSSIFVKLFKETPRIQKFFAKFANVAVDSLAGNAEYEKQVALVADRLDTMISAM DDKLQLLGNINYMRYTHTERGIPRGPWEDFGRLLLDSLAANGVSSDDLDSWKGVLA

^^^ ^ ^ VFVNGVSPRKD [0092] SEQ ID NOS: 7 Daphnia magna Hemoglobin chain 4 [0093] SLVGNAEYEKQIALVADRLDTMISAMDDKLQLLGNINYMRYTHTERGIPRAP WGDFSRLLLDVLGSKGVSTDDLDSWKGVLAVFVNGVSPRN [0094] SEQ ID NO: 8 Carcinus maenas Hemoglobin [0095] MGAVLSVVWGWLSPGTQVGAVTFPEEGSLGPEADVPDKTTGLTLRHRTAIYR TWDLVRPNPKLHGINLFLTMFQEEPVLQTRFKGFAGKSIEELKNSKRLAAHGTTVVM AITAMVDNLEDVSVLVELLKNTGANHRDRGVPKGDFELLAPVLVRFLKDNLGSAWS PVAEEAWTQAMKVINAVIFTSYDA [0096] SEQ ID NO: 9 Metacarcinus magister hemocyanin [0097] MKLLVLFALVAAAAAWPNFGIMADSAGGAFDAQKQHDVNSALWKVYEDIQ DPHLIQLSQNFDPLSGHYDDDGVAAKRLMKELNENRLLKQNHWFSLFNTRQREEAL MLYDVLEHSTDWSTFAGNAAFFRVSMNEGEFVYALYAAVIHSELTQHVVLPPLYEV TPHLFTNSEVIQEAYKAKMTQTAAKIESHFTGSKSNPEQRVAYFGEDIGMNTHHVTW HLEFPFWWDDAHENHHIERKGESFFWVHHQLTVRFDAERLSNYLDPVDELHWDDVI HEGFVPHTMYKYGGYFPSRPDNVHFEDVDGVARVRDMLILESRIRDAIAHGYVTGR DGSIISISDSHGIDVLGDVIESSLYSPNPEYYGALHNTAHMMLGRQGDPHGKFDLPPG VLEHFETATRDPAFFRLHKYMDNIFREHKDSLTPYTKDELEFPGVKIDSISLSNRLETY FEDFEYSLINAVDDTTKVADVPISTIVKRLAHKDFTVESVITNNNEHEVMATVRIFAW PMYDNNDVEFSFNDGRWNAIEMDKFWIKLASGTNTITRSSKDSSITVPDVPSFQDLIE KTKEAVSSGSELHLEEYVSGLGLPNRFLLPKGTTQGMKFHLVVFVSDGAKDAAIDGL HENTSFNHYGAHNGKYPDKRPHGYPMDRRVDDKRIFSGVTNIKCMNVKVYHVETQ [0098] SEQ ID NO: 10 Carcinus maenas hemocyanin [0099] MGAVLSVVWGWLSPGTQVGAVTFPEEGSLGPEADVPDKTTGLTLRHRTAIYR TWDLVRPNPKLHGINLFLTMFQEEPVLQTRFKGFAGKSIEELKNSKRLAAHGTTVVM AITAMVDNLEDVSVLVELLKNTGANHRDRGVPKGDFELLAPVLVRFLKDNLGSAWS PVAEEAWTQAMKVINAVIFTSYDA [0100] SEQ ID NO: 11 Human Myoglobin [0101] MGLSDGEWQLVLNVWGKVEADIPGHGQEVLIRLFKGHPETLEKFDKFKHLKS EDEMKASEDLKKHGATVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISE CIIQVLQSKHPGDFGADAQGAMNKALELFRKDMASNYKELGFQG

^^^ ^

Claims

^ MS 1. A method of repelling an arachnid or an insect from a subject or a fabric comprising applying a composition to the subject or the fabric, wherein the composition comprises one or more of hemoglobin, hemocyanin and myoglobin, and wherein the application repels the arachnid or the insect as compared to a control. 2. The method of claim 1, wherein the composition comprises hemoglobin and hemocyanin. 3. The method of claim 1, wherein the composition comprises hemocyanin and myoglobin. 4. The method of claim 1, wherein the composition comprises myoglobin and hemoglobin. 5. The method of claim 1, wherein the composition comprises hemoglobin. 6. The method of any one of claims 1, 2, 4 and 5, wherein the hemoglobin comprises one or more of an ^1, ^2, ^1 and ^2 subunit. 7. The method of any one of claims 1, 2 and 4-6, wherein the hemoglobin comprises one or more of SEQ ID NO: 1 and SEQ ID NO:2, or a sequence having at least 80% identity with SEQ ID NO: 1 and SEQ ID NO:2, or a fragment SEQ ID NO: 1 or SEQ ID NO:2. 8. The method of any one of claims 1-7, wherein the composition is applied to the subject’s skin. 9. The method of any one of claims 1-7, wherein the composition is applied to the fabric. 10. The method of any one of claims 1-9, wherein the arachnid is repelled. 11. The method of any one of claims 1-10, wherein the arachnid is a tick. 12. The method of claim 11, wherein the tick is a black legged tick. 13. The method of any one of claims 1-12, wherein the subject is a human. 14. The method of any one of claims 1-13, wherein the composition further comprises a pharmaceutically acceptable carrier and/or solvent. 15. The method of claim 14, wherein the pharmaceutically acceptable carrier comprises glycerol, water, petroleum jelly, aloe vera, coconut oil, and/or avocado oil.

^^^ ^ ^ 16. The method of any one of claims 1-15, wherein the arachnid or insect is repelled in an amount of at least 50% as compared to a control. 17. The method of any one of claims 1-15, wherein the arachnid or insect is repelled in an amount of at least 75% as compared to a control. 18. The method of any one of claims 1-15, wherein the arachnid or insect is repelled in an amount of at least 90% as compared to a control. 19. A composition comprising two or more of hemoglobin, hemocyanin and myoglobin. 20. The composition of claim 19, further comprising a pharmaceutically acceptable carrier. 21. The composition of claim 20, wherein the pharmaceutically acceptable carrier comprises glycerol, water, petroleum jelly, aloe vera, coconut oil, and/or avocado oil. 22. The composition of any one of claims 19-21, wherein the composition comprises hemoglobin and hemocyanin. 23. The composition of any one of claims 19-21, wherein the composition comprises hemocyanin and myoglobin. 24. The composition of any one of claims 19-21, wherein the composition comprises myoglobin and hemoglobin. ^ ^

^^^ ^