Compounds for reducing lactation and improving health

Abstract

Preferably, the compound comprises di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide, dibenzyl disulfide, diethyl sulfide, di-n-propyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide and di-n-propyl sulfone. The compound is selected from dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfinate (PTSO) and di-n-propyl thiosulfinate (PTS).

Description

Technical Field

[0001] The present disclosure relates to compounds for reducing lactation, improving health, reducing the risk or occurrence of intramammary infection, dry-off related stress, dry-off related inflammation and / or dry-off related infection. Preferred compounds include di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide, dibenzyl disulfide, diethyl sulfide, di-n-propyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfonate (PTSO) and di-n-propyl thiosulfinate (PTS). Background of the Invention

[0003] In the modern dairy industry, lactating animals go through a controlled milking and pregnancy cycle with significant overlap between the two phases. The "dry period" is usually induced between 40-70 days before expected parturition. The dry period is the transition period from the end of one lactation cycle to the beginning of a new lactation cycle after parturition. The purpose of the dry period is to stop milk production. Cows at this stage of the lactation cycle are called "dry cows." The dry period allows the mammary glands to recover, any intramammary infections to be treated, and allows healthy cows to provide high-quality milk in the new lactation cycle. The dry period also gives the cows the opportunity to produce colostrum for the calves after birth.

[0004] During the dry period, changes occur within the mammary gland that are important for the regeneration of new mammary tissue in preparation for lactation. The dry period also provides cows with an opportunity to eliminate pathogens within the udder that cause mastitis ((Boutinaud.M,Isaka N.,Gandemer E.,Lamberton P.,Wiart S.,De Prado AI,Sordillo LM,Lollivier V.2020). "Inhibiting prolactin by cabergoline accelerates mammary gland remodelling during the early dry period in dairy cows".Journal of Dairy Science.100(12):9789–9798)).

[0005] During the 12 to 24 hours of the non-lactation period, milk protein levels and expression of cell survival genes decrease, leading to a loss of epithelial cells. Changes in intracellular processes and gene regulation lead to a reduction in milk production until all milk production by mammary epithelial cells ceases (Hurley, WL (1989). Mammary Gland Function During Involution and the Declining Phase of Lactation. Journal of Dairy Science. 72 (6): 1637–1646). In addition, the content of milk-specific components such as lactose and fat also decreases, resulting in a rapid overall decline in milk production. After this, the mammary gland remains in a non-lactating state. After the calf is born and before lactation, the parenchymal tissue and other breast tissues in the mammary gland will redevelop. Colostrum is produced at the end of this stage.

[0006] The "dry day" is the day when the farmer stops milking and the cows begin the dry period. After drying off, the mammary gland continues to synthesize and secrete milk, which leads to increased intramammary pressure (pressure), which may cause pain and discomfort to animals (such as cows). 16 hours after drying off, milk accumulates in the mammary alveoli and ducts, causing udder swelling. About 16-18 hours after drying off, intramammary pressure increases rapidly, milk leakage and a mild inflammatory response occur. Evidence of inflammation includes a transient increase in blood flow, an increase in the number of neutrophils in the milk and changes in tight junctions. The intramammary pressure reaches a peak 2 days after drying off, then decreases, but still exists 4 or 6 days after the sudden dry off. During the sudden dry off period, the teat may open due to high udder pressure, which may cause milk leakage. In this case, the teat may become a portal for harmful microorganisms, resulting in (subclinical) mastitis. The formation of a keratin plug in each teat duct is an important natural defense mechanism against intramammary infection. However, not all cows will form such a keratin plug during the dry off period.

[0007] Generally, cows are abruptly stopped from being milked when their milk production drops below 15-20 litres per day. Abrupt drying is the most common method of drying up, used by 75% of dairy farms in the United States. With this method, milking is abruptly stopped on a day determined by the expected calving date and the corresponding length of the dry period. Abrupt drying is generally recommended for cows that produce less than 15-18kg of milk on the dry day. Since the density of milk is approximately 1.032kg / litre, this equates to 14.5-17.5 litres.

[0008] The advantages of sudden drying over other drying methods are convenience and labor saving. The disadvantages of sudden drying are an increased risk of mastitis due to a greater risk of milk leakage and intraudder pressure during drying, which is most likely to occur in high-producing cows. For example, at dry-off, the chance of mastitis increases by 77% for every 5kg increase in milk production above 12.5kg. Although all cows experience increased udder pressure after drying off, high-producing cows have the highest udder pressure and low-producing cows have the lowest udder pressure. This can lead to udder swelling and increase the chance of mastitis. In addition, sudden drying of cows producing more than 25 to 30kg / day of milk may result in reduced lying time within 3 days after drying off. Reduced lying time can increase acidosis and lameness due to reduced rumination and salivation rates. Blanket dry cow treatment (BDCT) (all quarters / cows receive antimicrobial therapy regardless of infection status) is often recommended to reduce the risk of intramammary infection in cows that are dried off suddenly, especially if milk production is high at dry-off.

[0009] For cows producing more than 15 litres of milk per day, dry-off management therapies can be used before dry-off. For example, gradual milking is a method that reduces milk production before dry-off by reducing the frequency of milking. This results in reduced milk secretion and stimulates mammary gland involution. Several studies have investigated the effect of milking frequency before dry-off on udder health. The advantages of gradual milking over abrupt dry-off are that it reduces the risk of new intramammary infections during the dry-off period and after calving due to reduced milk production, reduced risk of leaking milk and reduced intramammary pressure at dry-off. Many studies support gradual milking before dry-off to reduce milk production, but a reported disadvantage of this approach is that it slows down the formation of keratin plugs at the end of the teat. Therefore, cows that are milked gradually should be milked at least once a day to reduce the risk of mastitis. Another disadvantage of gradual milking is that the udder volume is reduced more slowly and an inflammatory response can occur, which negatively affects the cow's comfort. Cows also spend less time lying down when milking less frequently: the latter is a sign of discomfort. However, there is evidence that this practice may still cause some discomfort due to breast enlargement.

[0010] Another method of reducing milk production is through gradual feeding. In 2016, the USDA reported that 82% and 18% of dairy cows in the United States were dried off suddenly and gradually, respectively. With this method, the transport of glucose to the mammary gland is slowed by feeding, which reduces milk production. Gradual feeding can be done in a variety of ways, such as stopping feeding concentrates (14 days before dry-off), reducing dry matter intake (14 days), giving a low-energy diet (7 days before dry-off), or stopping feeding hay (5 days before dry-off). Feeding only hay may have negative effects on the health of dairy cows, such as decreased heart rate, increased plasma cortisol concentrations, and increased somatic cell counts at dry-off. While the gradual feeding method is effective in reducing milk production before weaning, extreme metabolic stress caused by feed restriction during the gradual feeding period should be avoided.

[0011] The advantage of gradual feeding is reduced milk production at weaning, which is associated with a reduced risk of new intramammary infections. Gradual feeding may be preferred over gradual milking due to lower milk production. The reduction in milk production with gradual feeding is caused by reduced nutrients to the mammary gland rather than by a mechanism of udder engorgement caused by udder pressure. Cows with reduced dry matter intake have been reported to leak less than half the amount of milk compared to cows that dry off suddenly.

[0012] Therefore, it is assumed that the risk of mastitis caused by leaking milk with gradual feeding is lower compared to gradual milking. However, this approach also has disadvantages. Gradual feeding by reducing dry matter intake increases the stress level of the cows. This stress can lead to impaired immunity and negative energy balance. In addition, if nutrient deficiencies are excessive and gestational undernutrition lasts for more than 80 to 90 days, it can have a negative impact on calf birth weight. Another disadvantage of gradual feeding is the additional labor requirement. Therefore, gradual feeding is less feasible on smaller dairy farms, where feeding cows with multiple total mixed rations is often problematic.

[0013] Despite extensive research, it is difficult to develop universal management methods that reduce milk production without any adverse effects on animal health or welfare (Martin et al., 2020. Automated gradual reduction of milk yield before dry-off: Effects on udder health, involution and inner teat morphology. Livestock Science, 233, 103942).

[0014] The reduction of milk production in mammals can be a painful process. This is especially true for mammals that experience this process frequently. Drying off in dairy cows is a dangerous, painful and stressful period. The mammary glands continue to secrete milk during the early involution period, leading to increased udder pressure and milk leakage. This can lead to an increased risk of intramammary infection in dairy cows, followed by (subclinical) mastitis, discomfort, pain and stress. The higher the milk production of dairy cows, the higher the risk of mastitis and the greater the pain and stress.

[0015] Udder health plays a vital role for dairy animals both from a hygienic and economic perspective. Mammary gland infections, i.e. mastitis, in dairy animals, such as dairy cows, have a major economic impact on the dairy industry worldwide. Several factors are known to disrupt the balance at the udder level, thereby impairing the ability of dairy animals to kill microorganisms that cause mastitis. As a result, host response mechanisms may not trigger an effective defense response to eliminate invading pathogens, leading to bacterial colonization of the udder and the occurrence of clinical or subclinical mastitis. Bacterial colonization in the udder of dairy cows, especially the formation of bacterial reservoirs, is often difficult to prevent and control, leading to infections, which usually require the use of antibiotics for treatment.

[0016] At dry-off, involution (the change of the mammary gland from lactating to non-lactating state) is caused by the sudden cessation of milk letdown. Once milk has accumulated in the udder after the dry-off day, new intramammary infections are very likely to develop. The reason for this susceptibility may be that the nipple duct is shortened due to udder pressure and the keratin plug has not yet fully formed.

[0017] During the dry period of dairy cows, about 10% to 17% of the udder compartments will develop new intramammary infections, leading to (subclinical) mastitis. (Pantoja et al. 2009. Somatic cell count status across the dry period as a risk factor for the development of clinical mastitis in the subsequent lactation. Journal of dairy science, 92 (1), 139-148). Infection during this period leads to a higher chance of mastitis in the subsequent lactation period (Capuco et al. 1997. A study of the incidence and significance of intramammary enterobacterial infections acquired during the dry period. Journal of dairy science, 83 (9), 1957-1965). Due to mastitis, milk production is reduced and milk quality deteriorates, resulting in relatively high economic losses (Hertl et al., 2014. Pathogen-specific effects on milk yield in repeated clinical mastitis episodes in Holstein dairycows. Journal of dairy science, 97(3), 1465-1480) and veterinary costs. Therefore, it is very important to prevent intramammary infection.

[0018] Treatment with topical antibiotics (AB) at the beginning of the dry period can reduce (subclinical) mastitis, but rarely resolves the problem because bacteria form biofilms when they are dormant, which makes them relatively insensitive to antibiotics. Scherpenzel et al. found that the incidence of clinical mastitis was 1.7 higher in subdivisions that were not dried off with antibiotics than in subdivisions that were dried off with antibiotics (Scherpenzeel et al., 2014. Evaluation of the use of dry cow antibiotics in low somatic cell count cows. Journal of Dairy Science, 97(6), 3606-3614). Since many countries only allow the use of antibiotics for therapeutic purposes to prevent microbial resistance to antibiotics, other methods must be found for high-quality dry period management to reduce the incidence of mastitis in the subsequent lactation period.

[0019] Dry cows are important to the future profitability of a dairy farm. Proper care, feeding and management of dry cows helps to increase milk production and udder health in the next lactation. Conversely, improper dry cow management can result in a 1,100 litre reduction in milk production. Therefore, proper care during the dry period is essential to achieve a productive and profitable dairy herd.

[0020] The human lactation cycle begins at conception. Delivery of the placenta triggers the transition to milk secretion, and continued mild synthesis requires regular milk ejection. The lactation cycle ends when the baby is weaned.

[0021] It is an object of the present disclosure to provide compounds and therapies for reducing lactation in mammals and for drying up of lactating dairy animals. Summary of the invention

[0022] The present invention provides compounds of formula I for use in treatment, particularly di-n-propyl disulfide. In particular, such compounds can be used to reduce lactation, prevent intramammary infection, and reduce stress, inflammation or infection risk during weaning / drying off. The following are preferred embodiments of the present disclosure.

[0023] 1. Compounds of Formula I

[0024]

[0025] Formula I,

[0026] Where R 1 and R 2 Independently selected from the group consisting of:

[0027] C 1-4Alkyl and phenyl; wherein, preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl or n-butyl; wherein preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted, or a composition comprising the compound is used for reducing lactation in a mammal.

[0028] 2. The compound or composition for use according to clause 1, wherein the mammal is a ruminant, preferably a cow.

[0029] 3. Compounds of Formula I

[0030]

[0031] Formula I,

[0032] Where R 1 and R 2 Independently selected from the group consisting of:

[0033] C 1-4 Alkyl and phenyl; wherein preferably the C 1-4 The alkyl group is methyl, ethyl, n-propyl or n-butyl; wherein preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted, or a composition comprising the compound is used for prophylactic treatment of intramammary infection and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection.

[0034] 4. The compound or composition of clause 3, wherein the mammal abruptly or gradually stops milking and the compound is administered prior to or on the day of cessation of milking.

[0035] 5. Compounds of formula I

[0036]

[0037] Formula I,

[0038] Where R 1 and R 2 Independently selected from the group consisting of:

[0039] C 1-4 Alkyl and phenyl; wherein preferably the C 1-4 The alkyl group is methyl, ethyl, n-propyl or n-butyl; wherein preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted, or compositions comprising the compounds are useful for promoting the health and welfare of lactating mammals.

[0040] 6. A compound or composition for use according to any one of the preceding clauses, wherein R 1 and R 2 same.

[0041] 7. A compound or composition for use as described in any of the preceding clauses, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, di-n-butyl disulfide and diphenyl disulfide.

[0042] 8. A compound or composition for use as claimed in any one of the preceding clauses, wherein the use is combined with dry cow therapy.

[0043] 9. A compound or composition for use as claimed in any one of the preceding clauses, further comprising administering a prolactin inhibitor, such as cabergoline, quinagolide; casein hydrolysate; or an acidogenic mineral bolus.

[0044] 10. A compound or composition for use according to any one of the preceding clauses, wherein the use further comprises the administration of an antibiotic, antifungal or anti-inflammatory agent.

[0045] 11. A method comprising administering to a mammal a pharmaceutical composition, a veterinary composition or a functional food composition comprising a compound of formula I

[0046]

[0047] Formula I,

[0048] Where R 1 and R 2 Independently selected from the group consisting of:

[0049] C 1-4 Alkyl and phenyl; wherein preferably the C 1-4 The alkyl group is methyl, ethyl, n-propyl or n-butyl; wherein preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted and are administered to a lactating mammal. Preferably, the method is used to reduce lactation in a mammal, and administration of the compound or composition results in reduced lactation. Preferably, the method is used to preventively treat intramammary infections in mammals and / or reduce the occurrence of dry milk-related stress, dry milk-related inflammation or dry milk-related infections in mammals, and administration of the compound or composition can preventively treat intramammary infections and / or reduce the occurrence of dry milk-related stress, dry milk-related inflammation or dry milk-related infections in mammals. Preferably, the method is used to promote the health and welfare of lactating mammals, and administration of the compound or composition can improve or maintain the health and welfare of lactating mammals.

[0050] 12. A method or compound or composition for use as claimed in any preceding clause, wherein the compound is formulated as a singledose unit comprising at least 50 grams, preferably at least 70 grams of the compound, and the composition is administered to a pregnant ruminant.

[0051] 13. A method or compound or composition for use as claimed in any preceding clause, comprising selecting a pregnant cow producing at least 10 litres of milk per day and administering the composition to the cow.

[0052] 14. A method or compound or composition for use as claimed in any preceding clause, wherein the mammal is abruptly or gradually stopped from milking and the compound is administered prior to or on the day of cessation of milking.

[0053] 15. A pharmaceutical or veterinary composition or a functional food composition comprising a compound of formula I

[0054]

[0055] Formula I,

[0056] Where R 1 and R 2 Independently selected from the group consisting of:

[0057] C 1-4 Alkyl and phenyl; wherein preferably the C 1-4 The alkyl group is methyl, ethyl, n-propyl or n-butyl; wherein preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted.

[0058] 16. The composition of clause 15, wherein the compound is formulated as a single dosage unit comprising at least 50 grams of the compound, preferably at least 70 grams of the compound.

[0059] 17. A composition according to clause 15 or 16, wherein R 1 and R 2 same.

[0060] 18. The composition of clause 17, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, di-n-butyl disulfide and diphenyl disulfide.

[0061] 19. A composition according to any one of clauses 15 to 18, further comprising a dry lactating agent, preferably selected from a prolactin inhibitor, such as cabergoline, quinagolide; a casein hydrolysate; or an acidogenic mineral bolus.

[0062] 20. The composition of any one of clauses 15-19, wherein the composition further comprises PTSO.

[0063] 21. The composition of any one of clauses 15-20, wherein the composition is a gel capsule.

[0064] The following are also preferred embodiments of the present disclosure.

[0065] 1. Compounds of formula II:

[0066]

[0067] Formula II, wherein

[0068] R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl;

[0069] Q 1 Selected from the following group: -SS-, -S-, -SSS-, -S(O) 2 -, -S(O)-S-, and -S(O) 2 -S-;

[0070] Provided that the compound of formula II is not diphenyl disulfide;

[0071] Among them, preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups;

[0072] Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl;

[0073] Preferably, the C 1-4 Alkyl and benzyl groups are unsubstituted;

[0074] or a composition comprising the compound, for

[0075] a) reduce lactation in mammals;

[0076] b) prophylactically treating intramammary infection in a mammal and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; and / or

[0077] c) promote the health and welfare of lactating mammals.

[0078] 2. Compounds of Formula I

[0079]

[0080] Formula I,

[0081] Where R1 and R 2 Independently selected from the group consisting of:

[0082] C 1-4 Alkyl and benzyl; wherein, preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted, or a combination of the compounds described herein is used

[0083] a) reduce lactation in mammals;

[0084] b) prophylactically treating intramammary infection in a mammal and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; and / or

[0085] c) promote the health and welfare of lactating mammals.

[0086] 3. The compound or composition for use according to embodiment 1 or 2, wherein the mammal is a ruminant, preferably a cow.

[0087] 4. A compound or composition for use as described in any one of the preceding embodiments, wherein R 1 and R 2 same.

[0088] 5. A compound or composition for use as described in any of the preceding embodiments, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide and dibenzyl disulfide.

[0089] 6. The compound or composition for use according to any one of the preceding embodiments, wherein the compound is di-n-propyl disulfide.

[0090] 7. A compound or composition for use as described in any one of embodiments 1 or 3-4, wherein the compound is selected from di-n-propyl sulfide, diethyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfonate (PTSO) and di-n-propyl thiosulfinate (PTS).

[0091] 8. A compound or composition for use as in any one of the preceding embodiments, wherein the use is combined with dry cow therapy.

[0092] 9. The compound or composition for use according to any of the preceding embodiments, further comprising administering a prolactin inhibitor, such as cabergoline, quinagolide; casein hydrolysate; or an acid-producing mineral pill.

[0093] 10. A compound or composition for use according to any one of the preceding embodiments, wherein the use further comprises administering an antibiotic, an antifungal agent, or an anti-inflammatory agent.

[0094] 11. A method for reducing lactation in a mammal; prophylactically treating intramammary infection in a mammal and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; or promoting the health and welfare of a lactating mammal, comprising administering to the mammal a pharmaceutical or veterinary composition or a functional food composition comprising a compound of formula II

[0095]

[0096] Formula II, wherein

[0097] R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl;

[0098] Q 1 Selected from the following group: -SS-, -S-, -SSS-, -S(O) 2 -, -S(O)-S-, and -S(O) 2 -S-;

[0099] Provided that the compound of formula II is not diphenyl disulfide;

[0100] Among them, preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups;

[0101] Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl;

[0102] Preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

[0103] 12. The method of claim 11, wherein the compound is a compound of formula I

[0104]

[0105] Formula I,

[0106] Where R 1 and R 2 Independently selected from the group consisting of:

[0107] C 1-4 Alkyl and benzyl; wherein, preferably, the C1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

[0108] 13. The method or compound or composition for use according to any one of the preceding embodiments, wherein the compound is formulated as a single dosage unit comprising at least 50 grams, preferably at least 70 grams of the compound, and the composition is administered to a pregnant ruminant.

[0109] 14. The method or compound or composition for use as in any one of the preceding embodiments, comprising selecting a pregnant cow producing at least 10 liters of milk per day, and administering the composition to the cow.

[0110] 15. The method or compound or composition for use as in any one of the preceding embodiments, wherein the mammal is abruptly or gradually stopped from milking and the compound is administered prior to or on the day of stopping milking.

[0111] 16. A pharmaceutical or veterinary composition or a functional food composition comprising a compound of formula II

[0112]

[0113] Formula II, wherein

[0114] R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl;

[0115] Q 1 Selected from the following group: -SS-, -S-, -SSS-, -S(O) 2 -, -S(O)-S-, and -S(O) 2 -S-;

[0116] Provided that the compound of formula II is not diphenyl disulfide;

[0117] Among them, preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups;

[0118] Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl;

[0119] Preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

[0120] 17. The composition of embodiment 16, wherein the compound is a compound of formula I

[0121]

[0122] Formula I,

[0123] Where R 1 and R 2 Independently selected from the group consisting of:

[0124] C 1-4 Alkyl and benzyl; wherein, preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

[0125] 18. The composition of embodiment 16 or 17, wherein the compound is formulated as a single dosage unit comprising at least 50 grams of the compound, preferably at least 70 grams of the compound.

[0126] 19. A composition according to any one of claims 16 to 18, wherein R 1 and R 2 same.

[0127] 20. The composition of claim 19, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide, dibenzyl disulfide, diethyl sulfide, di-n-propyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfate (PTSO) and di-n-propyl thiosulfinate (PTS).

[0128] 21. The composition of embodiment 19, wherein the compound is di-n-propyl disulfide.

[0129] 22. The composition of any one of embodiments 16-21, further comprising a dry milk agent, preferably selected from a prolactin inhibitor, such as cabergoline, quinagolide; casein hydrolysate; or an acid-producing mineral bolus.

[0130] 23. The composition of any one of embodiments 16-22, wherein the composition is a gel capsule.

[0131] Such compositions can be used in any of the methods or uses disclosed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0132] Figure 1Average milk production of 8 placebo cows of Example 1. At t=13 days, cows were treated with placebo capsules.

[0133] Figure 2 .Leakage. The two lines above represent the leakage mentioned in the previous literature (dark gray line) and the control group in this experiment (gray line), where the milk production of cows exceeded 12.5 kg / day. The bars show the percentage of cows that leaked milk at each fixed time after the sudden dry-off.

[0134] Figure 3 Udder pressure. The upper dark grey line and the lower grey line represent the udder pressure described in the previous literature and the control observations in this experiment, respectively, where the milk production of cows at dry-off exceeded 12.5 kg / day. The lower dark grey line represents the average udder pressure of cows treated with synthetic DPD at each fixed time after sudden dry-off.

[0135] Figure 4 . Stress. The upper line shows the expected stress in cows from the literature (grey line) and the control group of this experiment (dark grey) that produce more than 12.5 kg / day of milk at dry-off. The lower line shows the average stress in DPD-treated cows at each fixed time after the sudden dry-off.

[0136] Figure 5 . Milk leakage. The bars show the percentage of cows that leaked milk at each fixed time after the dry-off for cows treated with DPD-enriched OE. The line shows the expected amount of milk leakage.

[0137] Figure 6 Udder pressure. The lower line shows the average udder pressure at each fixed time after the dry-off. The upper line shows the expected udder pressure for cows producing more than 12.5 kg / day of milk at dry-off.

[0138] Figure 7 . Stress. The lower line shows the average stress behavior level of cows treated with DPD-enriched OE at each fixed time after dry-off. The upper line represents the expected stress for cows producing more than 12.5 kg / day of milk at dry-off.

[0139] Figure 8 . Udder pressure. The left column shows the udder pressure before the last milking, and the line below shows the udder pressure (g / m 2 The upper line is the data of the control group. The right column shows the udder pressure before the last milking, and the upper line shows the udder pressure (g / m 2 ).

[0140] Fig. 9 .DPD treatment and the occurrence of mastitis. Legend: vid-DPD-enriched OE treatment group; dol-DC liquid treatment group; AB-antibiotic treatment. DETAILED DESCRIPTION

[0141] The present disclosure provides compounds that can be used in a variety of methods. It will be clear to those skilled in the art that when referring to the use, administration, etc. of a compound, this also includes the use, administration, etc. of a composition comprising the compound.

[0142] The present disclosure provides compounds that can be used to reduce lactation in mammals and compositions comprising the compounds disclosed herein. As used herein, "lactation" refers to the secretion of milk by the mammary gland. Therefore, the compounds disclosed herein can be used to reduce milk production. In some embodiments, compared with the milk production before treatment, milk production is reduced by at least 10%. In some embodiments, the above comparison is performed between the day before treatment and the day after treatment.

[0143] The compounds disclosed herein and compositions comprising the compounds are useful for the prophylactic treatment of intramammary infections, such as mastitis.

[0144] High milk production before the dry period is one of the important factors for the potential occurrence of mastitis. One study showed that the probability of new (subclinical) mastitis in cows with a milk production of more than 5 kg before the start of the dry period was three times that of cows with lower milk production (Dingwell et al., 2001. Impact of milk production and important management factors on the process of dry-off inlactating dairy cows, Dairy Day 2001, p. 27). Another study showed that for every 5 kg increase in milk production above 12.5 kg, the probability of environmental (subclinical) mastitis at calving increased by 77% (Rajala-Schultz et al., 2005. Association between milk yield at dry-off and probability of intramammary infections at calving. Journal of Dairy Science, 88 (2), 577-579). The higher the milk production, the greater the udder pressure, and the more likely leakage (Summers et al., 2004. Influence of feeding level after drying off on incidence of mastitis and keratin plugformation in dairy cows. Proceedings of the New Zealand Society of Animal Production 2004, Vol 64). Leakage during the dry period leads to a four-fold higher chance of clinical mastitis during the dry period (Schukken et al., 1993. A randomized blind trial on dry cow antibiotic infusion in a low somatic cell count herd. Journal of Dairy Science, 76(10), 2925-2930).One study showed that cows with high milk production before the start of the dry period had a higher proportion of open teat ducts in the second and third weeks of the dry period and a higher incidence of (subclinical) mastitis after calving compared with cows with lower milk production at the start of the dry period (Odensten et al., 2007. Metabolism and udder health at dry-off in cows of different breeds and production levels. Journal of Dairy Science, 90(3), 1417-1428). Sealed or blocked teat ducts are less susceptible to mastitis during the dry period than open teat ducts (Williamson et al., 1995. The prophylactic effect of a dry-cow antibiotic against Streptococcus uberis. New Zealand veterinary journal, 43(6), 228-234). Therefore, the rapid formation of a keratin plug after the start of the dry period is very important to prevent new intramammary infections during the dry period (Lacy-Hulbert et al., 1999. Sealing of the bovine teat canal after drying off. In Proceeding-New Zealand Society of Animal Production (Vol. 59, pp. 198-200). This keratin plug forms a physical barrier to prevent bacteria from entering the teat canal in this way. Low milk production at the beginning of the dry period helps to minimize milk leakage and promotes the formation of a keratin plug. This can prevent intramammary infections during the dry period and the resulting new (subclinical) mastitis.

[0145] As used herein, "prophylactic treatment of intramammary infection" refers to reducing the likelihood of intramammary infection of the breast or udder and / or reducing the severity and / or duration of symptoms of infection. Preferably, the treatment is of a mammal. Preferably, the treatment results in maintaining the health of the individual.

[0146] The compounds disclosed herein and compositions comprising the compounds are useful for reducing the occurrence of infections, particularly dry-off-associated infections or weaning-associated infections.

[0147] Infections can be caused by a variety of pathogens, the most prominent of which are bacteria and viruses. Other microorganisms have also been implicated in udder infections, such as microalgae such as Prototeca spp., mycoplasmas, viruses, yeasts, and fungi. The mammalian host responds to infection with an innate response that typically includes inflammation, followed by an adaptive response.

[0148] The most important microorganisms that can invade the udder and cause bovine mastitis are Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactia, Streptococcus dysgalactiae, as well as Serratia marcescens, Leptospira spp., Pseudomonas spp., Brucella spp., Escherichia coli, Klebsiella spp., Mycobacterium spp. and other facultative pathogenic Enterobacteriaceae. Examples of viruses that can invade the mammary gland and can directly or indirectly cause mastitis are bovine herpesvirus 1, bovine herpesvirus 2, vaccinia, bovine viral diarrhoeavirus cowpox, pseudocowpox, vesicular stomatitis, foot-and-mouth disease viruses and bovine papillomaviruses, bovine immunodeficiency virus, parainfluenza 3 and bovine leukaemia virus infections, which can play an (indirect) role in the etiology of bovine mastitis. These viruses can induce nipple lesions, such as teat duct lesions, leading to a decrease in the natural defense mechanisms of the udder, indirectly leading to bovine mastitis caused by bacterial pathogens. In addition, when the milk is dried off, there is an increased likelihood that potentially pathogenic yeasts, protozoa and fungi, such as Candida spp., Cryptococcus spp., Rhodotorula spp., Stephanoascus spp., Trichosporum spp., Kodamaea spp., Kloeckeria spp., Aspergillus spp. and Neospora, will enter the teat.

[0149] The compounds and compositions disclosed herein containing the compounds can be used to reduce the occurrence of inflammation, in particular inflammation associated with dry milk or inflammation associated with weaning. Inflammation is part of a complex biological response of body tissues to (harmful) stimuli (such as pathogens, less milking or no milking), and is a protective response involving immune cells and molecular mediators. The function of inflammation is to eliminate pathogens.

[0150] During the dry period or weaning period, infections (such as udder infections in dairy cows) may occur, leading to tissue inflammation. Activated immune cells and inflammatory responses can also damage tissues, such as mammary tissue. Therefore, inhibiting the inflammatory response can prevent or reduce damage to tissues. For example, the mammary gland is less damaged and the cow's milk production recovers faster.

[0151] The compounds disclosed herein and compositions comprising the compounds are useful for reducing the occurrence of stress, particularly stress associated with drying off or stress associated with weaning.

[0152] While these uses are intended for prevention, those skilled in the art recognize that prevention is generally not a 100% reduction, but rather a reduction in likelihood compared to an individual not treated with a compound of the invention.

[0153] Compounds disclosed herein and compositions comprising the compounds can be used to promote the health and / or welfare of lactating mammals, particularly in the case of needing to reduce milk production. As mentioned above, the process of weaning and drying off may be painful, tense, and has the risk of intramammary infection. Compounds disclosed herein have an active role in weaning and drying off. In addition, although not wishing to be bound by theory, compounds disclosed herein may have an impact on serotonin, oxytocin, prolactin and / or dopamine.

[0154] As further described herein, the compound or composition may be administered to a mammal in combination with cessation of milking. In particular, the compound or composition may be administered prior to or on the day that the mammal (abruptly or gradually) ceases milking.

[0155] The compounds disclosed herein and compositions comprising the compounds can be administered to any mammal, preferably a lactating mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human mammal. Preferably, the mammal is a ruminant (such as a cow and a goat), more preferably a cow. In some embodiments, the mammal is in pregnancy. In a specific embodiment, the compound is administered to a lactating mammal during pregnancy to reduce lactation before delivery. In some embodiments, the mammal is a cow, and preferably a cow with a milk production greater than 10 L / day, preferably 11.5 L / day, is selected for treatment.

[0156] The disclosure also provides for the in vitro use of the compounds disclosed herein for studying the effects on mammals, preferably ruminants, cows or human mammary tissue. Preferably, the method is used to reduce milk formation of mammary tissue cultured on a surface. In some embodiments, the method comprises contacting tissue attached to a surface with a composition disclosed herein.

[0157] The present disclosure relates to compounds of formula I

[0158]

[0159] Formula I,

[0160] Where R 1 and R 2 Independently selected from the group consisting of:

[0161] C 1-4 In some embodiments, R 1 and R 2 Independently selected from the following group: C 1-4 In a preferred embodiment, R 1 and R 2 same.

[0162] The present disclosure also relates to compounds of formula II

[0163]

[0164] Formula II, wherein

[0165] R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl;

[0166] Q 1 Selected from the following group: -SS-, -S-, -SSS-, -S(O) 2 -, -S(O)-S-, and -S(O) 2 -S-;

[0167] Provided that the compound of formula II is not diphenyl disulfide. In some embodiments, the compound of formula II is not diethyl sulfide. In a preferred embodiment, R 1 and R 2 Independently selected from the following group: C 1-4 In a preferred embodiment, R 1 and R 2 same.

[0168] As used herein, "alkyl" refers to a saturated aliphatic hydrocarbon group. Unless otherwise specified, an alkyl group may be straight chain or branched. Preferably, the alkyl group is straight chain. As used herein, an alkyl group may be substituted or unsubstituted. Preferably, the alkyl group is unsubstituted. Preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl.

[0169] As used herein, "substituted" means that the group contains one or more substituents. Preferably, the substituents are independently selected from halogen, -C(O)OH, -C(O)NH 2 、-OH、=O、C 1-3 Alkoxy, -NH 2 、-NO 2 、-SO 3 H and CF 3 Preferably, the halogen is selected from the group consisting of -Cl, -F, -Br and -I. Most preferably, the halogen is -Cl. In a preferred embodiment, the groups disclosed herein contain at most three substituents, more preferably at most two substituents, most preferably at most one substituent. Preferably, the C 1-4 The alkyl and phenyl groups are unsubstituted. 1-4 The alkyl and benzyl groups are unsubstituted.

[0170] In a preferred embodiment, the compound of formula I is selected from the following group: di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, di-n-butyl disulfide and diphenyl disulfide. In a preferred embodiment, the compound of formula I or formula II is selected from the following group: di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide and dibenzyl disulfide. In a preferred embodiment, the compound of the present invention is selected from the compounds described in Table 18.

[0171] In a preferred embodiment, the compound of Formula I or Formula II is di-n-propyl disulfide.

[0172] In a preferred embodiment, the compound of formula II is selected from the group consisting of diethyl sulfide, di-n-propyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfinate (PTSO) and di-n-propyl thiosulfinate (PTS).

[0173] In some embodiments, the compound is obtained from a natural source (such as a plant). Compounds can be extracted from plant materials in a variety of ways. The appropriate method depends on the chemical nature of the compound. For example, extraction can be performed using a non-polar solvent and then a solvent with increased polarity. Compounds can also be prepared synthetically.

[0174] In a preferred embodiment, the compound of formula I or formula II is selected from the group consisting of di-n-propyl disulfide (DPD; CAS No. 629-19-6), dimethyl disulfide (DMDS CAS No. 624-92-0), diethyl disulfide (CAS No. 110-81-6), diisopropyl disulfide (CAS No. 4253-89-8), di-n-butyl disulfide (CAS No. 629-45-8), diphenyl disulfide (CAS No. 882-33-7) and dibenzyl disulfide (CAS No. 150-60-7). In a preferred embodiment, the compound of formula II is selected from the group consisting of diethyl sulfide (CAS No. 352-93-2), di-n-propyl sulfide (CAS No. 111-47-7), diisopropyl sulfide (CAS No. 625-80-9), di-n-butyl sulfide (CAS No. 544-40-7). 1), diphenyl sulfide (CAS No. 139-66-2), dibenzyl sulfide (CAS No. 538-74-9), di-n-propyl trisulfide (CAS No. 6028-61-1), di-n-propyl sulfone (CAS No. 598-03-8), dibenzyl thiosulfinate (CAS No. 16302-98-0), dibenzyl thiosulfinate (CAS No. 16601-40-4), diisopropyl thiosulfinate (CAS No. 10027-69-7), di-n-propyl thiosulfinate (PTSO; CAS No. 1113-13-9) and di-n-propyl thiosulfinate (PTS; CAS No. 1948-52-3). These compounds are commercially available, for example, from Sigma-Aldrich.

[0175] In a preferred embodiment, the compound is di-n-propyl disulfide (DPD). As described in the examples, surprisingly, DPD reduces milk production without significant toxic side effects. 200 grams of di-n-propyl disulfide were administered to dairy cows without showing any adverse effects. In the examples described herein, 80 ml of pure di-n-propyl disulfide reduced milk production as well as di-n-propyl disulfide at a dose as low as 20 ml (Example 2). The density of di-n-propyl disulfide is 0.96 g / ml (20° C.). Therefore, a dose of 80 ml of pure di-n-propyl disulfide is equivalent to 77 grams.

[0176] Table 17 (Example 8) shows exemplary doses of compounds of formula II for dairy cows in milliliters (ml) or grams (g). In a preferred embodiment, the minimum dose of the compound of formula II is at least 30% of the exemplary dose. For example, the minimum dose of diethyl disulfide is at least 18.9 ml. In a preferred embodiment, the minimum dose of the compound of formula II is at least 50% of the exemplary dose. For example, the minimum dose of diethyl disulfide is at least 31.5 ml. In a preferred embodiment, the minimum dose of the compound of formula II is at least 80% of the exemplary dose. For example, the minimum dose of diethyl disulfide is at least 50.4 ml. In some embodiments, the maximum dose is 200% or 300% of the exemplary doses shown in Table 17.

[0177] The actual dosage level of the compounds described herein can be varied to obtain an amount of active ingredient that is effective to achieve the desired therapeutic response for a particular individual, composition, and mode of administration without causing toxicity to the individual. The selected dosage level will depend on a variety of factors, including the activity of the particular compound, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, the compounds and / or materials used in combination, the age, sex, weight, condition, overall health and past medical history of the individual being treated, and similar factors well known in the medical field. A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the effective amount of the desired compound.

[0178] In some embodiments, at least 0.2g, preferably at least 0.5g of the compounds disclosed herein are given to mammals every day. In an exemplary embodiment, at least 50 grams, preferably at least 60 grams, more preferably at least 70 grams of compounds disclosed herein (preferably DPD) are provided to dairy cows. In some embodiments, at least 75 grams or at least 77 grams of compounds disclosed herein (preferably DPD) are provided to dairy cows. In some embodiments, 50-200 grams, preferably 70-200 grams, more preferably 75-200 grams of compounds disclosed herein (preferably DPD) are provided to dairy cows. Preferably, at least 77 grams of compounds disclosed herein (preferably DPD) are provided to dairy cows. Preferably, about 77 grams of compounds disclosed herein (preferably DPD) are provided to dairy cows. These dosages can be provided as single dose units.

[0179] It is clear to those skilled in the art that lower doses of the compound can be given to smaller animals. The examples describe administration of tablets containing 77 grams of DPD to dairy cows. Since the average weight of a dairy cow is about 650 kg, this is equivalent to a dose of about 118 mg DPD / kg. It is known to those skilled in the art that smaller animals have higher metabolic rates and therefore smaller animals require larger drug doses calculated by body weight. Dose conversion between animals and between humans and animals is reviewed in the following literature: Nair and Jacob (J Basic Clin Pharm. March 2016-May 2016; 7(2):27–31) and Holliday et al. (1967 The Relation of Metabolic Rate to Body Weight and Organ Size. Review. Pediat. Res. 1: 185-195).

[0180] Although not wishing to be bound by theory, the present disclosure provides that compounds disclosed herein can produce favorable effects after a single administration. In a preferred embodiment, the effect can be achieved by providing a single administration of compounds disclosed herein. As shown in the examples, a single administration results in a (reversible) reduction in milk production. When combined with other strategies (such as dry milk or weaning), milk production can be further and more permanently reduced. In an exemplary embodiment, compounds disclosed herein are provided to lactating, pregnant cows, and milking is not performed until the calves are born. Once new calves are born, milk production will resume.

[0181] The disclosure also provides multiple administrations. For example, the composition can be provided more than once per day, daily, weekly or monthly. In an exemplary embodiment, the composition can be provided once a week until milk production stops or significantly reduces. In some embodiments, the composition can be provided once every 3-4 days or every 2 days.

[0182] These compounds are particularly useful when provided systemically (e.g., orally). As further disclosed herein, the compounds are preferably provided as pharmaceutical or veterinary compositions or functional foods. Those skilled in the art will appreciate that such compositions are suitable for administration to humans and other animals.

[0183] In some embodiments, the compositions disclosed herein are provided as functional food compositions. As used herein, the term "functional food" refers to those prepared not only for its nutritional properties, but also for achieving a specific function (such as improving health or reducing the risk of infectious diseases). Such functional foods may also be referred to as dietary supplements or (animal) food additives. For this reason, bioactive compounds such as minerals, vitamins, fatty acids, bacteria with beneficial effects, dietary fiber and antioxidants, etc., may be added thereto. Such food may be in any form suitable for oral administration, such as liquid, gel, powder, pill, tablet or gel capsule.

[0184] Functional foods can also include animal digests, such as any material produced by chemical and / or enzymatic hydrolysis of clean and undecomposed animal tissues. Functional foods can also include dry brewer's yeast, such as a dry, inactive agent as a byproduct of the brewing industry. It has been found that animal digests and dry brewer's yeast can enhance the palatability of functional foods. When present in functional foods, animal digests account for about 10% to about 90% of the functional food, and dry brewer's yeast accounts for about 1% to about 30% of the functional food.

[0185] In some embodiments, the compositions disclosed herein are provided as medicines or veterinary compositions. In some embodiments, the present invention provides compositions comprising compounds disclosed herein and at least one pharmaceutically acceptable carrier, diluent and / or excipient. (See, for example, Remington: The Science and Practice of Pharmacy, Alfonso R. Gennaro (ed.) Mack Publishing Company, April 1997). As used herein, the term "pharmaceutically acceptable" refers to those compositions or reagents, materials or compositions and / or combinations of their dosage forms within the scope of reasonable medical judgment, which are suitable for contacting human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications compatible with a reasonable benefit / risk ratio. In addition, as used herein, the term "pharmaceutically acceptable diluent or carrier" refers to pharmaceutically acceptable materials, compositions or carriers, such as liquid or solid fillers, diluents, excipients, solvents or encapsulating materials, which participate in carrying or transporting peptides from one organ or part of the body to another organ or part of the body.

[0186] In some embodiments, the composition comprises at least 10% by weight of a compound disclosed herein. Preferably, the composition comprises at least 10% by weight of DPD. In some embodiments, the composition comprises at least 40% by weight of a compound disclosed herein, preferably at least 50% by weight of a compound disclosed herein. In some embodiments, the composition comprises at least 60% by weight of a compound disclosed herein, preferably at least 70% by weight of a compound disclosed herein. In some embodiments, the compound disclosed herein is the sole active ingredient of the composition.

[0187] In some embodiments, the composition does not include one or more of the following compounds: isopentanol, dimethylthiophene, 2-undecanone, tridecane, 2-hexyl-5-methyl-3(2H)-furanone, 2-tridecanone, methyl palmitate, ethyl palmitate, methyl lineolate, and ethyl oleate. In some embodiments, the composition does not include

[0188] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight of isoamyl alcohol,

[0189] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight of dimethylthiophene,

[0190] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight, of 2-undecanone,

[0191] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight of tridecane,

[0192] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight, of 2-hexyl-5-methyl-3(2H)-furanone,

[0193] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight, of 2-tridecanone,

[0194] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight, of methyl palmitate,

[0195] more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.3% by weight, preferably more than 0.1% by weight, of ethyl palmitate,

[0196] - more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight of methyl linoleate, and / or

[0197] - more than 10% by weight, preferably more than 1% by weight, more preferably more than 0.1% by weight of ethyl oleate.

[0198] The composition can be administered by any suitable route and mode. It will be appreciated by those skilled in the art that the route and / or mode of administration will vary depending on the desired result. The composition can be formulated according to conventional procedures and administered by any route, such as parenteral or enteral administration. Preferably, the composition is administered orally.

[0199] In some aspects, oral administration includes administering the composition in combination with the animal's feed, water or medication. In some aspects, oral administration includes applying the composition in a gel or viscous solution to an animal's body part or spraying the composition on an animal's body part, wherein the animal ingests the composition by licking. In some embodiments, when the mammal is a ruminant, the composition is injected into the rumen.

[0200] The composition can be in any suitable form, such as liquid, semisolid and solid dosage forms. The composition can be in the form of tablets, capsules, powders, granules, lozenges, liquid preparations. Preferably, the composition is suitable for oral administration. Such oral compositions include tablet or pill preparations. As used herein, a pill refers to a single dose of material that can be swallowed. In some embodiments, the composition is an intraruminal bolus.

[0201] In an exemplary embodiment, the composition is a gel capsule comprising a compound disclosed herein. Suitable gel capsules are known in the art, including bovine, porcine and fish gelatin capsules. Preferably, the gel capsule contains at least 50 grams of DPD, preferably 50-200 grams of DPD.

[0202] In some embodiments, the treatments disclosed herein (such as administration of a compound disclosed herein) can be combined with other therapies (such as dry cow therapy).

[0203] For example, treatment can be combined with abrupt or gradual withdrawal of milk. Abrupt withdrawal means stopping active milk removal by pumping, expressing milk, or natural feeding (such as breastfeeding). Gradual withdrawal means reducing milk removal. This can mean reducing the frequency or amount of milk removal.

[0204] In an exemplary embodiment, the compounds disclosed herein are provided to lactating cows within 24 hours, preferably within 12 hours, before the "dry day". In another exemplary embodiment, the compounds disclosed herein are provided to lactating animals, preferably dairy animals such as cows or goats, 12-24 hours before the "dry day". In such an embodiment, starting from the dry day, the animal is no longer milked until the calf is born (a new lactation cycle begins). In some embodiments, the compound or composition is administered within 4-8 hours after the last milking. Then the animal is no longer milked until after delivery.

[0205] In another embodiment, the treatment can be combined with gradual feeding. Gradual feeding is a method of reducing milk production by reducing the rate at which glucose is delivered to the mammary gland. Gradual feeding can be achieved in a variety of ways known to those skilled in the art. For example, removing concentrates from the diet for 14 days before drying off, reducing dry matter intake for 14 days, giving a low-energy diet for 7 days before drying off, or removing hay 5 days before drying off (for a review of dry-off methods and gradual feeding, see https: / / edis.ifas.ufl.edu / pdf / AN / AN36000.pdf). For example, the average dry matter intake of a dry cow is about 21 kg / day, but this amount can be halved if gradual feeding is used.

[0206] In some embodiments, the use of the compounds disclosed herein can be combined with one or more other treatments used to reduce milk production, prevent or treat mastitis or other infections, or prevent or treat inflammation. Such combinations can be formulated into a single composition, thereby containing multiple active ingredients, or these combinations can be administered separately. For example, DPD can be administered a few days before or after administration of one or more other products.

[0207] In some embodiments, a combination therapy is provided, which comprises a compound disclosed herein and a prolactin inhibitor. Prolactin, i.e., prolactin, is an important protein hormone known for its role in causing mammals to produce milk. By administering a prolactin inhibitor, the formation or release of prolactin can be affected, thereby inhibiting the formation of milk. Examples of prolactin inhibitors include quinagolide and cabergoline.

[0208] Cabergoline is an ergot derivative that inhibits prolactin release. In one study, the overall effect of cabergoline administration was a reduction in prolactin secretion, udder swelling and milk leaking, and improved lying time. A single injection of cabergoline (5.6 mg) during the dry period reduced prolactin for up to 8 days after dry-off and reduced milk production by 28% in goats and 22% in cows.

[0209] Cabergoline is a ergoline derivative that stimulates the dopamine D2 receptors of the prolactin-secreting cells of the pituitary gland, thereby inhibiting the release of prolactin. A single intramuscular injection of cabergoline was sufficient to reduce the prolactin concentration in plasma during the first week of the dry-off period compared to the untreated placebo group. In addition, a decrease in lactose concentration and an increase in lactoferrin concentration in milk were detected (Boutinaud et al., 2016. Cabergoline inhibits prolactin secretion and accelerates involution in dairycows after dry-off. Journal of dairy science, 99 (7), 5707-5718) Cabergoline for intramuscular injection is marketed under the trade name Velactis. Cows treated with Velactis had a lower incidence of mastitis 7 days after calving than the control group, at 20.5% and 26%, respectively. It was found that the milk leakage rate was also lower than that of the control group, at 2% and 11%, respectively. Furthermore, behavioral observations showed that Velactis administration reduced udder pain compared to the control group. However, the drug is no longer permitted for use in the EU following reports of adverse events. 319 cows experienced serious adverse events following treatment, such as lying down (208 cows) and death (71 cows) (European Medicines Agency, 2016).

[0210] Another prolactin inhibitor is quinagolide: this is a dopamine receptor agonist. It stimulates dopamine D2 receptors and inhibits the release of prolactin from the anterior pituitary. A group of early lactating cows received 1 mg of quinagolide intramuscularly every day for 9 weeks. This reduced milking-induced prolactin release, but did not reduce basal prolactin concentrations. In addition, milk production was reduced by 5.3 kg / day in the last 4 weeks of treatment compared to the placebo group (Lacasse et al., 2011. Effect of the prolactin-release inhibitor quinagolide on lactating dairy cows. Journal of dairy science, 94(3), 1302-1309). Another group of cows received intramuscular injections of 44 mg twice a day from 5 days before dry-off to 13 days after dry-off. During this treatment period, prolactin concentrations in blood, milk, and mammary secretions decreased. In addition, milk production decreased compared to the placebo group, which was 18 kg / day and 25 kg / day, respectively. In this experimental design, another group of cows was fed only hay instead of the usual lactation diet, and it was found that prolactin concentrations in the blood and milk were also reduced. Milk production was reduced by 10 kg / day. The difference between cows treated with quinagolide and cows fed only hay was in the metabolites in the blood. The first group showed an increase in blood glucose levels, but the levels of other metabolites were not affected. In the second group, a decrease in blood glucose levels, a decrease in the levels of most amino acids, and an increase in the blood concentrations of β-hydroxybutyrate and non-esterified fatty acids were observed. Feed restriction causes a significant drop in milk production, but quinagolide can be a good alternative to reduce milk production without interfering with metabolism (Ollier, Zhao, and Lacasse, 2014. Effects of feed restriction and prolactin-release inhibition at drying off on metabolism and mammary gland involution in cows. Journal of dairy science, 97(8), 4942-4954). One disadvantage of using quinagolide is the need for frequent dosing. Because of the additional cost, dairy farmers may decide to only give prolactin inhibitors to cows that produce more than 15 to 18 kg (33 to 40 pounds) of milk per day at dry off.

[0211] Acidogenic mineral boluses contain anionic salts that can cause temporary metabolic acidosis. One study showed that cows given two 196g boluses produced 2.6kg / day less milk the day after treatment, compared to 1.2kg for cows given one bolus and 0.23kg / day for a control group. Another experiment showed that cows given two boluses reduced feed intake over a 3-day period after treatment. The mineral composition of the oral bolus is NH 4 Cl 10.4%, calcium chloride 51.9%, calcium sulfate 20.8%, water 12.6% and coating material (mono- and di-glycerides of fatty acids esterified with acetic acid) 4.3%, each dose weighs 196g. Each dose of pills provides about 20g (about 10.4% of the total weight of the pills) of NH 4 Cl. Milk production decreased on days 2 and 3 after treatment. In addition, the pH of urine from treated cows was lower than that from control cows. In addition, the results showed that cows treated with 2 boluses had lower udder pressure compared to the control group, but there was no change in the incidence of milk leakage between the treatment and control groups. Therefore, potential mastitis-causing microorganisms could still have entered. Maynou et al. (2018, Effects of oral administration of acidogenic boluses at dry-off on performance and behavior of dairy cattle. Journal of dairyscience, 101(12), 11342-11353) mentioned two possible explanations for the reduced feed intake in the first 3 days after treatment. The first attributed the observation to changes in the acid-base status of the cows' rumen. The second explanation is that this could be due to potential damage to the rumen wall. The decrease in milk production is partly due to the reduced feed intake. β-Hydroxybutyrate concentrations were lower compared to the control group. This is due to reduced feed intake, which in turn leads to lower levels of butyrate metabolized in the rumen wall (Maynou et al., 2018).

[0212] Another ingredient that stimulates dry milk is casein hydrolysate (CNH). Casein hydrolysate causes loss of mammary gland tight junction integrity (Shamay et al., 2003. Infusions of casein hydrolysate into the mammary gland disrupt tight junction integrity and induce involution in cows. Journal of Dairy Science, 86(4), 1250-1258; Ponchon et al., 2014. Effects of intramammary infusions of casein hydrolysate, ethylene glycol-bis(β-aminoethylether)-N,N,N′,N′-tetraacetic acid, and lactose at drying-off on mammary gland involution. Journal of dairy science, 97(2), 779-788). Shamay et al. demonstrated that CNH infusion is an effective and gentle method for downregulating milk production during lactation and drying off chronically infected and therapy-resistant single udder compartments.

[0213] Seeth et al. (2016. Drying-off single udder quarters of dairy cattle during lactation using caseinhydrolysate. Milk Science International 69:23-26) showed that infusion of casein hydrolysate was an effective and gentle method for downregulating milk production during lactation and drying off chronically infected and treatment-resistant single udder quarters if each treatment udder quarter received 6 intracisternal infusions of casein hydrolysate over 3 treatment days. In the pH dissertation (2019) by Justine Elena Britten (Evaluation of Casein Hydrolysate as an Alternative Dry-Off. Treatment and Milk Quality Management Tool in Dairy Cows Utah State University. Dissertations Graduate Studies 5-2019), the use of casein hydrolysate was evaluated and concluded to be a promising tool for dry-off. Animals treated with casein hydrolysate did not show any signs of discomfort or pain. These studies suggest that intramammary infusion of casein hydrolysate is safe for dairy cows, has some efficacy against mastitis, and may have added value in reducing mastitis in both lactating and dry cows. This was confirmed by Britten et al. (2021) (Comparison of Bovine Mammary Involution and Intramammary Infections Following Intramammary Treatment with Casein Hydrolysate and Other Conventional Treatments at Dry-Off. Animals 2021, 11(8), 2360).

[0214] In some embodiments, a combination therapy is provided, which comprises a compound disclosed herein and an anti-inflammatory agent. Such drugs can be used to inhibit inflammatory reactions and reduce tissue damage, such as loss of mammary tissue during dry-off. Anti-inflammatory agents include, for example, nonsteroidal anti-inflammatory agents (cox / lox inhibitors), such as ibuprofen, paracetamol, aspirin, diclofenac, ketoprofen, tolmetin, etodolac and fenoprofen. Natural anti-inflammatory agents can also be used, such as curcumin, ginger, spirulina, cayenne pepper (Cayenne), cinnamon, cloves, sage, rosemary, black pepper, natural aspirin, frankincense (Boswelia), bloodroot grass (Sanguinaria) and / or green tea.

[0215] In another embodiment, the treatment can be combined with a teat sealant. Internal teat sealants are often given to cows after dry cow treatment. Internal teat sealants are usually a paste injected into each teat to create a physical barrier for organisms. Teat sealants can reduce new infections in the days after dry off, when the keratin plug (a natural barrier composed of a waxy substance) at the end of the teat has not yet fully formed.

[0216] Teat sealants can be given internally or externally. Antibiotics are often used in treatments that are inserted into the teat before the sealant is applied. Afterwards, teat dips or sprays can be applied after milking. Cows should stay in a clean area for at least 30 minutes after treatment and avoid long walks after drying off. Once the cow is dry, the condition of the udder should be checked regularly for signs of inflammation and infection.

[0217] In some embodiments, the treatment disclosed herein (e.g., administration of a compound disclosed herein) can be combined with an antimicrobial agent such as an antibiotic or antifungal agent. While not wishing to be bound by theory, the present disclosure provides that a compound disclosed herein can prevent intramammary infection, while an antimicrobial agent can work on remaining infections or help prevent new infections.

[0218] Exemplary antimicrobial agents that can be used for combined treatment include antifungal agents such as miconazole, ketoconazole, econazole, terbinafine, ciclopirox, tolnaftate, sertaconazole, sulconazole, amphotericin B, cholorxylenol, clioquinol, butenafine, naftifine, nystatin and clotrimazole. Exemplary antibiotics include penicillins, tetracyclines, cephalosporins, quinolones, lincomycins, macrolides, sulfonamides, glycopeptides, aminoglycosides and carbapenems.

[0219] One current approach to preventing intramammary mastitis infections (dry cow therapy) is to inject antibiotics and / or teat sealants into the udder compartments of dairy cows on the dry day with the goal of preventing and treating dry period infections. The use of antibiotics can reduce existing infections and new infections at the beginning of the dry period. Dry cow therapy is categorized as “catch-all” and “selective” therapy. Catch-all dry cow therapy, which is used for the majority of treatments (93% of cows) in the United States (USDA, 2016). In catch-all therapy, antibiotics are injected into all compartments of all cows in the herd, regardless of intramammary infection status. However, administering catch-all dry cow therapy to cows that do not need antibiotics may contribute to antimicrobial resistance.

[0220] In contrast, selective dry cow therapy means that antibiotics are only used for cows that are infected or at abnormally high risk for infection. Candidates for selective dry cow therapy are cows that have low somatic cell counts within 3 months before dry off. Herds with low rates of subclinical mastitis are good candidates for selective dry cow therapy.

[0221] The optimal conditions for drying off cows are known to those skilled in the art. It is important to minimize stress during the dry period because of the negative effects on appetite and immunity. Social stress can be reduced by avoiding herd changes to keep social hierarchies as independent as possible. Some farmers separate dry cows from other cows to ensure that the cows are no longer milked. Environmental conditions such as ventilation and temperature are also important. Cows are also usually provided with a modified diet during the dry period.

[0222] In a preferred embodiment, the compounds disclosed herein are used in combination with compounds of formula I and / or formula II disclosed in WO2021 / 182958, the entire contents of which are incorporated herein by reference.

[0223] As used in the literature, the compound of formula I (WO 2021 / 182958) is as follows:

[0224]

[0225] Wherein R1 and R2 are independently selected from: optionally substituted straight chain or branched alkyl, optionally substituted straight chain or branched alkenyl, optionally substituted straight chain or branched alkynyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl. Preferably, the compound of formula I is propyl propanethiosulfonate (PTSO).

[0226] As used in the literature, the compound of formula II (WO 2021 / 182958) is as follows:

[0227]

[0228] Wherein R3 and R4 are independently selected from: optionally substituted straight chain or branched alkyl, optionally substituted straight chain or branched alkenyl, optionally substituted straight chain or branched alkynyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl. Preferably, the compound of formula II is propyl propanethiosulfinate (PTS).

[0229] The combination of the compounds of the invention with the compounds of formula I (WO 2021 / 182958) or formula II (WO 2021 / 192958) makes it possible to treat existing infections and prevent new infections before drying off the milk.

[0230] In some embodiments, the compound of formula I (WO 2021 / 182958) and / or the compound of formula II (WO 2021 / 182958) is administered before the dry-off date, preferably one to two months before the dry-off date. The compounds of the present invention are administered according to the methods disclosed herein, for example, around the dry-off date.

[0231] The present invention further provides a kit comprising a first composition comprising a compound of formula I (WO 2021 / 182958) and / or a compound of formula II (WO 2021 / 182958) and a second composition comprising a compound of the present invention as disclosed herein.

[0232] As used herein, the articles "a" and "an" refer to one or more than one (ie, at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element.

[0233] When used in conjunction with numerical values ​​(about 10, approximately 10), the word "about" or "approximately" preferably means that the value may be 1% greater or less than 10 of the given value.

[0234] The compounds and compositions disclosed herein can be used as treatments and for therapeutic therapies, and thus can be used as medicaments and methods for preparing medicaments. In some embodiments, the methods provided by the present disclosure are not methods of treatment of the human or animal body, and / or do not include processes that alter human germline genetic characteristics.

[0235] wherein the cells are not human germ line cells.

[0236] All literature and patent references cited in this specification are hereby incorporated by reference in their entirety.

[0237] The present invention is further explained in the following examples. These examples are not intended to limit the scope of the present invention but are merely intended to illustrate the present invention.

[0238] Example

[0239] A variety of methods have been applied and studied in the field of drying off dairy cows. For example, Pattamaont et al. (2020. https: / / edis.ifas.ufl.edu / publication / AN360) reviewed the most commonly used methods and summarized their advantages and disadvantages. Since the incidence of new intramammary infections is 77% higher during the dry period compared to the lactation period, there is a strong need for new compounds, compositions and / or methods, or a combination of existing methods and new methods, to reduce the risk of new infections. The examples described below demonstrate the effect of a single oral dose of DPD on milk production.

[0240] Example 1. The range of effects of 40 ml and 80 ml doses of DPD (di-n-propyl disulfide) on milk production in dairy cows: Different doses of DPD were orally administered to dairy cows, and their effects on milk production were measured.

[0241] Study Design:

[0242] Fifteen cows from three different farms were used in Study 1 and 30 cows from four different farms were used in Study 2. The dry period for each cow began 40 to 60 days before calving. Cows were milked twice a day at fixed times, and cows with a milk production of >11.5 L / day in the 24 hours before DPD administration were selected. No additional measures were taken to stimulate dryness, such as changing the feeding diet or changing the milking frequency. Cows were milked twice a day. Cows were housed in free stables. Porcine gelatin capsules containing DPD or placebo gelatin capsules containing water were administered 7 to 14 days before the start of the dry period. Farmers were not informed which cows were treated.

[0243] In study 1, gel capsules containing 40 ml of DPD were orally administered via a bolus shooter, and the gel capsules in the placebo group contained tap water as described above. There were 8 cows in the placebo group and 7 cows in the treatment group.

[0244] In study 2, gelatin capsules contained 80 ml of DPD and gelatin capsules in the placebo group contained tap water. Nine cows were given a single dose of DPD and nine cows were given a single dose of placebo capsule 7 to 14 days before the start of the dry-off period. Cows were randomized to either the DPD group or the placebo group. Each farm independently dried off each group of cows at the same time, so that each farm had 50% of cows in the treatment group and 50% in the control group.

[0245] The capsules were given between morning and afternoon milking. At t=13 days, capsules containing DPD and placebo capsules were given.

[0246] The following hypotheses were tested and compared by statistical analysis: H0 was defined as no difference in relative milk production between the DPD bolus group and the placebo group before and after treatment, and H1 was defined as a difference in relative milk production between the DPD bolus group and the placebo group before and after treatment.

[0247] Results and discussion

[0248] Cow Description

[0249] In the treatment group, the mean age of the 15 cows was 5.5 years (standard deviation 2.1 years), the mean gestation period (parity) was 2.9 years (standard deviation 1.9 years), and the mean milk production was 20.2 liters per 24 hours (standard deviation 4.5 liters).

[0250] The mean age of the nine cows in the placebo group was 4.5 years (standard deviation 1.3 years), the mean gestation period was 2.8 years (standard deviation 1.2 years), and the mean milk production was 23 L / 24 h (standard deviation 4 L), which was determined 24 h before treatment.

[0251] Study on milk production of dairy cows treated with 1.40 ml DPD

[0252] The milk production of the cows showed high variability from day to day. This is common because cows can vary greatly in milk production. To illustrate the day-to-day variability in milk production, milk production of 4 cows in each treatment is presented in Table 1. A total of 8 cows were used in this study, so not all data are shown in this table.

[0253] Table 1 shows the milk production of 4 cows in the placebo group and the 40 ml DPD group.

[0254]

[0255] 251*: There are two different cows with the same numbers on their ears.

[0256] Figure 1 The average milk yield of 8 placebo cows is shown. The absolute milk yield per cow may vary significantly, but a clear downward trend was observed after treatment. Administration of capsules containing water showed no sudden change in the steady decline in milk yield (t=13 days). ND=Not Determined.

[0257] Data processing

[0258] Data processing was as follows: Based on the data obtained on days 0, 1, 2, 3...12, the expected milk production of each cow on days 13, 14, 15...20 was calculated by linear regression. At t=13, a placebo capsule or a capsule containing DPD was administered to the rumen. From t=13 onwards, the predicted data (without the administration of placebo or DPD capsule) were compared with the actual milk production after administration, and the milk reduction was calculated. This was done for all cows participating in the trial.

[0259] Thereafter, the mean milk yield reduction was calculated by comparing the expected milk yield obtained by linear regression with the actual milk yield obtained after treatment with placebo or DPD and calculating the difference between the observed and expected milk yields per day.

[0260] Table 2 shows the results for placebo-treated cows.

[0261] Placebo Group:

[0262]

[0263] Table 2. Mean reduction in milk yield (difference) in placebo-treated cows. "Difference" refers to the mean of the observed actual milk yield minus the expected milk yield obtained by linear regression.

[0264] This method was also applied to the group of cows treated with 40 ml DPD capsules and the corresponding 80 DPD capsules. This method for 40 ml DPD is given in Table 3.

[0265] 40ml DPD treatment group

[0266]

[0267] Table 3. Average milk yield reduction (difference) of cows treated with 40 ml DPD.

[0268] Afterwards, we tested whether the differences between treatment and placebo were significantly different. The null hypothesis was that the differences were zero. We calculated P values ​​for these differences using a permutation test (5000 samples). The results are shown in Table 4.

[0269]

[0270] Table 4. Significance of the effect of treatment with 40 ml DPD on milk production in cows compared with cows treated with placebo. Mean differences are expressed in litres of milk per cow per day. The "2.5% percentile" and "97.5% percentile" restrict the confidentiality interval of the null hypothesis.

[0271] It is clear from the P values ​​that treatment with 40 ml DPD did not result in a significant decrease in milk production. However, two cows showed a decrease in milk production after treatment, while the remaining two cows did not. It was observed that the variability of the decrease in milk production at lower DPD doses was greater than the variability of the decrease in milk production at higher DPD doses. Upon closer inspection, the work procedures during the low-dose experiments were not as organized in terms of the time of milking each day, and there was little supervision during the conduct of these experiments. This resulted in a relatively high standard deviation, which made the effect of low-dose DPD on milk reduction less obvious. Therefore, in Example 2, a dose-response curve of the administered dose of DPD and the milk production after administration was constructed under standardized and closely monitored experimental conditions. Therefore, the results regarding Example 2 are considered to be close to the actual situation.

[0272] Study on milk production of cows treated with 2.80ml DPD

[0273] A similar experiment was conducted as described above with 40 ml DPD administration, using 80 ml DPD instead. The results are as follows

[0274] As shown in Table 5.

[0275]

[0276] Table 5. Average milk yield reduction (difference) of cows treated with 80 ml DPD.

[0277] Thereafter, the difference between the 80 ml treatment and placebo was tested to see if it was significantly different. The null hypothesis was defined as the difference between the treated and placebo cows being zero. We calculated the P values ​​for these differences using a permutation test (5000 samples). The results are shown in Table 6.

[0278]

[0279] Table 6. Significance of the effect of treatment with 80 ml DPD on milk production in cows compared to cows treated with placebo. Mean differences are expressed in litres of milk per cow per day. The "2.5% percentile" and "97.5% percentile" restrict the confidentiality interval of the null hypothesis.

[0280] It is clear from the P values ​​that treatment with 80 ml DPD resulted in a significant decrease in milk production.

[0281] Results and discussion

[0282] Before treatment, there was no significant difference in relative milk production between the DPD and placebo groups. Milk production in cows treated with 80 ml DPD decreased significantly within 3 days of treatment compared with the placebo group (average 6 liters per cow per day), but after this period, milk production was found to recover.

[0283] Combining this treatment with other drying methods (e.g., sudden drying, gradual expression, gradual feeding, or other methods) is expected to reduce milk production and the risk of new udder infections and stress / pain.

[0284] Rajala Schultsz et al. (2005. Association between milk yield at dry-off and probability of intramammary infections at calving. Journal of Dairy Science, 88(2), 577-579) determined that for every 5 kg increase in milk production above 12.5 kg at dry-off, the probability of environmental IMI (intramammary infection) at calving increased by 77%. Therefore, treatment of cows with DPD significantly reduced the probability of environmental IMI at calving. This was confirmed by Odensten et al. (2007, Metabolism and udder health at dry-off in cows of different breeds and production levels. Journal of Dairy Science, 90(3), 1417-1428) who determined that cows producing more than 11.5 kg of milk per day in the week before weaning had a higher percentage of open teat ducts in the second (30%) and third (12%) dry weeks and a higher incidence of IMI after calving (21.8%) compared to cows producing less milk. When DPD treatment results in milk production below 11.5 kg / day, it is expected that the percentage of open teat ducts after calving will decrease and the incidence of IMI will also decrease. The study by Maynou et al. (2018) showed that the group that received two doses of acid-producing mineral boluses had reduced milk production on days 2 and 3 after treatment, with milk production on day 2 decreasing by 2.6 liters, compared to a decrease of 0.23 liters in the placebo group. Compared to this product, DPD showed a greater effect on day 2 after treatment, with a decrease of 7.7 liters per day in milk production observed compared to a decrease of 0.75 liters per day in the placebo group. This makes DPD a more effective dry milk product than acid-producing mineral boluses. In the study by Ollier, Zhao, and Lacasse (2014), treatment with quinagolide reduced milk production by 7 liters compared to the placebo group. The advantage of DPD over quinagolide is the simple mode of administration. Quinagolide must be injected frequently, while DPD is administered orally and a single dose is sufficient.

[0285] Example 2. Dose-Response DPD Treatment and Milk Production

[0286] In this example, the correlation between reduced milk production and different doses of synthetic DPD was determined.

[0287] Study design 27 dairy cows (Holstein Friesians) from 2 farms were used. The dry period for each cow began 40 to 60 days before calving. Cows were milked twice daily at fixed times, and cows with milk production >12.5 L / day in the 24 hours before DPD administration were selected. No additional measures were taken to stimulate dryness, such as changing the feeding diet or changing the milking frequency. Cows were milked twice daily. Cows were housed in free stables. Porcine gelatin capsules containing DPD or placebo gelatin capsules containing water were administered 7 to 14 days before the start of the dry period.

[0288] In this study, different doses of synthetic DPD (purity >98%) were given to dairy cows producing at least 12.5 kg of milk per day. The following doses were administered: 0, 10, 20, 50, 100, 150, 175, 200 ml of pure DPD. Three cows were used for each dose.

[0289] Milk production was monitored for 14 days before the different doses were given. The gel capsules were given orally with a pill booster and the gel capsules for the placebo group contained tap water. The capsules were given between morning and afternoon milking. At t=14 days, capsules containing DPD and placebo capsules were given and milk production was monitored for 5 days.

[0290] Results. Table 7 shows the milk production and the change in milk production (%) after administration of different doses of DPD. A positive value for "percent reduction" indicates a decrease in milk production, while a negative value indicates an increase in production. The reference treatment of administering water capsules instead of DPD capsules over the next five days did not have any effect on milk production. However, a dose of 20 ml of DPD significantly reduced milk production compared to pre-treatment milk production, and the reducing effect became stronger when more DPD was administered. No further decrease in milk production was observed when more than 80 ml was administered. In addition, the cows showed a relaxed and quiet behavior, and no changes in eating behavior were observed. However, when the dose exceeded 100 ml, the cows appeared to become more active.

[0291] Table 7. Milk production and reduction in milk production (expressed as a percentage of the reduction in milk production before treatment) after administration of different doses of DPD.

[0292]

[0293]

[0294] ND*: Not determined.

[0295] Discussion and conclusion. This study showed that a dose-response curve can be constructed between the dose of DPD and the decrease in milk production. The decrease in milk production became obvious when 20 ml or more of DPD was given. The maximum response was obtained after a dose of 80 ml or more of DPD. No adverse behavior of the cows was observed until 80 ml of DPD, but when 80 ml or more of DPD was given, the cows became restless and showed more active behavior. The effects of DPD disappeared on the 3rd and 4th days after treatment, which strongly suggests that the treatment did not cause damage to the (udder) health of the cows. On the contrary, DPD treatment provides the possibility of preventing or reducing milk leakage after sudden drying and preventing new mastitis infections. Secondly, the treatment can also reduce or prevent painful udder pressure associated with stress. Therefore, giving cows DPD during sudden drying is a valuable means to benefit the health of dairy cows and the profitability of farms.

[0296] Example 3. Effects of synthetic (purity>98%) DPD on udder pressure, stress behavior and milk leakage in high-producing dairy cows after sudden drying off

[0297] List of abbreviations: IMI: intramammary infection; sDCT: selective dry cow therapy; bDCT: total dry cow therapy; ITS: intramammary teat sealer; DIL: days in lactation; SCC: somatic cell count.

[0298] Scope: High-producing dairy cows that dry up suddenly are particularly susceptible to new infections, which are stressors such as discomfort caused by high udder pressure, leading to stress and milk leakage. New IMI (intramammary infection) is mainly caused by bacteria entering the udder through the teat canal. Different doses of DPD were given orally to dairy cows and their effects on discomfort and milk leakage were measured.

[0299] Study Design

[0300] Holstein Friesian cows with a milk production of at least 12.5 kg per day in the last lactation were selected. In the three studies of this example, these cows were treated with 80 ml of di-n-propyl disulfide (DPD) immediately after the last milking and then dried off. Due to the limited number of cows available, only study 3 had a control group. During the first 72 hours, cows were monitored for udder pressure, milk leakage, and stress every 12 hours after the last milking.

[0301] Animals and design

[0302] Forty-six dairy cows (Holstein Friesian cows) from seven farms in the Netherlands participated in a two-month field study. A description of some housing conditions and information about the cows is given in Table 8.

[0303] Table 8. Description of housing conditions and cows on the seven farms. All tested cows produced more than 12.5 kg of milk per day. Cows were dried off immediately after the last milking and moved directly to the dry group without any concentrate intake. Cows received ITS treatment. The dry period started 40 to 60 days before calving. All cows were housed in free stables.

[0304]

[0305] After the last milking, the cows were isolated from other lactating cows. Only in farms 2 and 6, the cows could hear the sound of the milking machine. After isolation, the cows were given 80 ml of synthetic DPD. Thereafter, the farmers monitored the cows for 72 hours and scored them every 12 hours after the sudden dry-off. The farmers visually scored milk leakage (yes / no-score), udder pressure and stress according to a standard scoring form (see Tables 9 and 10). Cow somatic cell counts were extracted from the last milking record before dry-off and the first milking record after calving (5-30 days of lactation).

[0306] Table 9. Description of udder condition of dairy cows and corresponding scores.

[0307]

[0308] Table 10. Description of stress levels and corresponding scores of dry cows.

[0309]

[0310] Data analysis: The control group was scored for milk leakage, udder pressure, and stress and compared with data previously provided in the literature. For milk leakage, the data were compared assuming that 24.5% of cows leaked milk after dry-off (De Prado-Taranilla et al., 2020. Incidence of milk leakage after dry-off in European dairy herds, related risk factors, and its role in new intramammary infections. Journal of Dairy Science, 103(10), 9224–9237). Udder pressure (Bertulat et al., 2013. Measurement of fecal glucocorticoid metabolites and evaluation of udder characteristics to estimate stress after sudden dry-off in dairy cows with different milk yields. Journal of Dairy Science, 96(6), 3774–3787) and stress (Chapinal et al., 2014. Changes in lying behavior after abrupt cessation of milking and regrouping at dry-off in free stall-housed cows: A case study. Journal of Veterinary Behavior, 9(6), 364–369) were converted into a scoring table as expected results (Table 11).

[0311] Table 11. Expected scores for breast pressure and stress.

[0312] Time after drying off 12h 24h 36h 48h 60h 72h Breast Pressure Prediction Score 3 4.2 5 5 4.5 3.8 Stress anticipation score 2.3 2.8 3.7 3.4 2.9 2.9

[0313] result

[0314] On the last day before dry-off, the average milk production of the 46 Holstein Friesian cows was 22.3 kg / day (production ranged from 15.6 kg / day to 29.7 kg / day). None of the cows experienced any adverse effects during the study. Figure 2 , 3 and 4 show the results for milk leakage, breast pressure, and stress.

[0315] The average SCC of the 46 cows at the last milking of the previous lactation was 89,000 cells / ml (range 45,000-106,000 SCC / ml) and at the first milking of the new lactation was 58,000 (36,000-112,000). None of the cows had milk fever at calving.

[0316] discuss

[0317] The milk reduction effect produced by administration of 80 ml of pure DPD was shown to be sufficient to successfully dry off high-producing cows. In previous studies, high milk production on the last day before drying off has been shown to cause udder pressure and milk leakage (Chapinal et al., 2014; Bertulat et al., 2013; De Prado Taranilla et al., 2020), and these observations are consistent with the results shown in the experimental control in this example. This study also showed that administration of the specified dose of synthetic DPD reduced udder pressure, stress, and milk leakage when drying off suddenly compared to the control group.

[0318] The purpose of administering DPD at the start of dry-off is to reduce the risk of new infection through the udder teat. Milk leakage is primarily caused by excessive udder pressure and is a common cause of new IMI at the beginning of the dry period (Klaas et al., 2005. Cow-Related Risk Factors for Milk Leakage. Journal of Dairy Science, 88(1), 128–136; Rajala-Schultz et al., 2005. Short Communication: Association Between Milk Yield at Dry-Off and Probability of Intramammary Infections at Calving. Journal of Dairy Science, 88(2), 577–579; Gott et al., 2016. Intramammary infections and milk leakage following gradual or abruptcessation of milking. Journal of Dairy Science, 99(5), 4005–4017). However, although this treatment reduced udder pressure and ITS, a low proportion of cows with leaking milk remained. However, this reduced the incidence of IMI (Example 4).

[0319] The dry period is the period when the dairy industry uses the most antibiotics, especially to prevent new IMI or treat subclinical mastitis (Kuiper et al., 2014. Antibiotic use in dairy herds in the Netherlands from 2005 to 2012. Sciencedirect.com.

[0320] https: / / www.sciencedirect.com / science / article / pii / S0022030215009054?ref=pdf_download&fr=RR-2&rr=738fe0be1d72b7d3). DPD is effective in reducing new infections during the dry period (as shown in Example 4), and DPD can reduce the need for long-acting antibiotics during the dry period and after calving.

[0321] in conclusion

[0322] DPD can be a useful additive to support cows after a sudden dry-off. DPD-treated cows showed significantly lower udder pressure compared to control cows. This was associated with a lower percentage of cows leaking milk and lower stress levels, and resulted in fewer new IMIs. This is a valuable positive contribution to the farm’s economy and the cow’s health.

[0323] Example 4 - Effect of DPD-enriched onion extract on udder pressure, stress behavior and milk leakage in high-producing dairy cows after sudden dry-off

[0324] Scope: In this example, onion extract was enriched with 80 ml of DPD to give an onion extract enriched with DPD (= DPD enriched OE). Two studies were conducted after a single dose of 150 ml of DPD enriched OE. After administration of 150 ml of DPD enriched OE, the milk was dried off suddenly and the effect on three dry-off variables was investigated, namely the effect on milk leakage (study 1), stress behavior and udder pressure (study 2).

[0325] Study 1. Effects on milk leakage and stress levels.

[0326] Study design. Cow description: Thirty-one dairy cows (Holstein Friesian cows) from six farms in the Netherlands participated in this field study. The dry period for each cow began 40 to 60 days before calving, corresponding to the optimal interval of dry period (Capuco et al., 1997. Mammary Growth in Holstein Cows During the Dry Period: Quantification of Nucleic Acids and Histology. Journal of Dairy Science, 80(3), 477–487). A description of some housing conditions and information about the dairy cows is given in Table 12.

[0327] Table 12. Description of the housing conditions and cows on the six farms. All the cows tested produced more than 12.5 kg of milk per day. The cows were dried off immediately after the last milking and moved directly to the dry group without any concentrate intake. The cows were treated with ITS (intramammary teat sealant). The dry period started 40 to 60 days before calving. All cows were housed in free stables.

[0328]

[0329]

[0330] The minimum milk production of the cows on the dry day is 12.5 kg / day. The cows are dried off suddenly after the last milking and transferred directly to the dry group.

[0331] After the last milking, cows were treated with ITS and treated with 150 ml of DPD-enriched OE. All cows were housed in free stalls separate from milking cows.

[0332] The farmers followed the cows for 72 hours and scored them every 12 hours after the sudden dry-off. The farmers had to visually score milk leakage (and give a qualitative yes / no score), udder pressure and stress according to the standard scoring table (see Tables 10 and 11 of Example 3). The somatic cell count values ​​of the cows were taken from the last milk record before dry-off and the first milk record after calving (lactation day 5-30).

[0333] Data processing. Due to the small number of cows available, data for the control group were obtained from the literature: udder pressure and stress scores were compared with the expected data described previously (Chapinal et al., 2014. Changes in lying behavior after abrupt cessation of milking and regrouping at dry-off in free stall-housed cows: A case study. Journal of Veterinary Behavior, 9(6), 364–369; Bertulat et al., 2013. Measurement of fecal glucocorticoidmetabolites and evaluation of udder characteristics to estimate stress after sudden dry-off in dairy cows with different milk yields. Journal of DairyScience, 96(6), 3774–3787). This is demonstrated to be a reliable method in Example 3. Regarding milk leakage, the data were compared with earlier studies showing that 24.5% of cows leaked milk after dry-off (De Prado-Taranilla et al., 2020. Incidence of milk leakage after dry-off in European dairy herds, related risk factors, and its role in new intramammary infections. Journal of Dairy Science, 103(10), 9224–9237).

[0334] H0: Cows will score the same or better in terms of reduced udder pressure, reduced stress levels, and reduced milk leaking when given DPD-enriched OE (containing 80% native DPD) at the beginning of the dry period. Due to the lack of a control group in this experiment, no statistical analysis was performed: the data obtained were compared with historical data obtained from the literature.

[0335] The following study was conducted to analyze whether 150 ml of DPD-enriched OE is effective in preventing stress, udder pain, and milk leakage after dry-off in high-producing dairy cows (>12.5 kg milk / day).

[0336] Study 2 - Breast pressure after dry-off

[0337] Study design. Animal description: In July 2022, six dairy cows (Holstein Friesian cows) from 1 farm in the Netherlands participated in this field study. The dry period for each cow started 40 to 60 days before calving, as this is the optimal interval before the start of the dry period. The minimum milk production of cows on the day of dry-off was 12.5 kg / day. Cows were dried off suddenly after the last milking and transferred directly to the dry group, where they could not hear the milking robot. After the last milking, cows with even numbers (2, 4, 6) were treated with ITS and 150 ml of DPD-enriched OE. The other three cows (control group) were treated with ITS only after the last milking. All cows were housed in free stables.

[0338] After 72 hours of dry-off, all cows were followed and udder pressure was measured with a penetrometer (Medista 5000 Digital Hardness Tester, supplied by Medista, 13 Rue du Bastringue 76440 Serqueux, La France). Cows were measured before and directly after the last milking (maximum and minimum stress were expected, respectively). In addition, udder pressure was monitored 12, 24, 36, 48, 60 and 72 hours after the last milking (Table 13). All cows were measured with a penetrometer in the left front subdivision 10 cm directly above the teat (the first measurement was marked with a red marker).

[0339] Table 13. Schedule for measuring breast pressure using a penetrometer

[0340]

[0341] H0: Cows treated with DPD-enriched OE will have lower udder pressure values ​​after dry-off compared to untreated cows. Results were statistically analyzed using t-tests for differences in means.

[0342] result

[0343] Study 1:

[0344] On the last day before dry-off, the average milk production of the 31 Holstein Friesian cows was 24.5 kg / day (production ranged from 17.7 kg / day to 29.9 kg / day). None of the cows experienced any adverse effects during the study. Figure 5 ,6 and 7 show the results for milk leakage, breast pressure, and stress.

[0345] The average SCC of the 46 cows at the last milking of the previous lactation was 84,000 cells / ml (range 31,000-97,000 cells / ml) and the average SCC at the first milking of the new lactation was 61,000 cells / ml (range 36,000-97,000 cells / ml). None of the cows had milk fever at calving.

[0346] Study 2

[0347] Udder pressure was measured in three experimental and three control cows after sudden dry-off. Two of the three control cows leaked milk after 36 hours, while none of the experimental cows treated with DPD-enriched OE leaked milk. Figure 8 shown.

[0348] from Figure 8 It can be seen that after administration of 150 ml of DPD-enriched OE, breast pressure was significantly reduced.

[0349] Discussion and Conclusions. In this example, it was clearly demonstrated that a single dose of DPD-enriched OE can reduce udder pressure, reduce milk leakage, and reduce stress behaviors. It is described in the literature that there is a positive correlation between milk leakage after sudden drying and the occurrence of mastitis. Since milk leakage is a result of teat duct opening after sudden drying, a large number of mastitis cases can be prevented, which is economically important. In addition, sudden drying is a painful process because high udder pressure leads to a large number of stress behaviors, so this method is very beneficial to the health of dairy cows.

[0350] Example 5. Dose-Response PTSO Treatment and Milk Production

[0351] This example shows that dipropylthiosulfonate (PTSO), a compound chemically related to DPD, exhibits the same or similar effects as DPD in reducing milk production. In this example, PTSO was used as a demonstrator compound. In addition, the correlation between different doses of PTSO and reduced milk production was determined.

[0352] Study design. Twelve dairy cows (Holstein Friesian cows) from 1 farm were used. The dry period for each cow began 40 to 60 days before calving. Cows were milked twice daily at fixed times, and cows with a milk production >12.5 L / day in the 24 h before PTSO administration were selected. No additional measures were taken to stimulate dryness, such as changing the feeding diet or changing the milking frequency. Cows were milked twice daily. Cows were housed in free stables. Porcine gelatin capsules containing PTSO or placebo gelatin capsules containing water were administered 7 to 14 days before the start of the dry period.

[0353] In this study, different doses of synthetic PTSO (purity >98%) were given to dairy cows with a daily milk production of at least 12.5 kg. The following doses were administered: 0, 10, 20, 40 ml of pure PTSO. Three cows were used for each dose.

[0354] Milk production was monitored for 14 days before the different doses were given. The gel capsules were given orally with a bolus booster and the gel capsules for the placebo group contained tap water. The capsules were given between morning and afternoon milking. At t=14 days, capsules containing PTSO and placebo were given and milk production was monitored for a further 48 hours.

[0355] Results. Table 1 shows the milk production and the change in milk production (%) after administration of different doses of PTSO. Positive values ​​of "percent reduction" indicate a decrease in milk production, while negative values ​​indicate an increase in production. The reference treatment of administering water capsules instead of PTSO did not show any effect after 24 and 48 hours. However, a dose of 20 ml of PTSO significantly reduced milk production compared to the milk production before treatment, and the reduction effect became stronger when more PTSO was administered.

[0356] Table 14. Milk production and changes in milk production (expressed as percentage change compared to pre-treatment milk production) after administration of different doses of PTSO.

[0357]

[0358] Discussion and conclusion. This study showed that there was a dose-response curve between the dose of PTSO and the decrease in milk production. The effect became evident when 20 ml or more of PTSO was given. The maximum response was obtained after a dose of 20 ml or more of PTSO. The study also showed that milk production fully recovered after 3 to 4 days, and the recovery rate depended on the dose of PTSO. This suggests that the milk-reducing effect of PTSO is reversible and does not cause any damage to the udder.

[0359] These treatments offer the potential to prevent or reduce milk leakage, thereby preventing new mastitis infections, and secondly, to reduce or prevent painful udder pressure associated with stress during the dry-off period. Therefore, giving cows PTSO during the dry-off period is a valuable tool that benefits cow health and farm profitability.

[0360] Example 6. Synthesis of di-n-propyl thiosulfinate

[0361] Scope: This method describes the synthesis of di-n-propylthiosulfinate. This compound is used to treat dairy cows to study its effect on milk production.

[0362] Study design. Under nitrogen atmosphere, in a three-necked flask, di-n-propyl disulfide (139.5 g, 1 eq., 928 mmol) was dissolved in 460 ml of dichloromethane (DCM) and cooled to about 0-2°C (internal). Meta-chloroperbenzoic acid (m-CPBA) (228.8 g, 70% Wt, 1.00 eq., 1.33 mol) was dissolved in DCM (2.3 L) and the solution was added dropwise to the cooled di-n-propyl disulfide solution over 6 hours while maintaining the temperature below 5°C. After complete addition, the reaction mixture was filtered and the solvent was evaporated under reduced pressure at about 30°C to give a colorless oil. The oil was dissolved in isopropyl acetate (IPAc) (1 L) and washed with NaHCO 3 The mixture was washed with aqueous solution (5%, 6x, 500 mL), water (2x300 mL) and brine (1x, 400 mL) and then with Na 2 SO 4 Drying, filtration and evaporation at about 30°C gave a yellow oil (144 g). Purification by automated column chromatography (1.6 kg silica, 0-7% IPAc:heptane, run time 5.5 h, 250 mL / min) gave PTS (122.4 g) as a yellow oil, which still contained some residual m-chlorobenzoic acid. This was redissolved in IPAc (1 L) and treated with NaHCO 3 The mixture was washed with aqueous solution (5%, 6x, 500 mL), water (2x300 mL) and brine (1x, 400 mL), and then with Na 2 SO 4 Drying, filtration and evaporation at about 30°C gave PTS (100 g, 65%) as a yellow oil, the structure of which was analyzed by 1H NMR and the purity by LCMS.

[0363] 1H NMR (400MHz, CDCl3): δ3.18-3.00 (m, 4H), 1.90-1.72 (m, 4H), 1.06 (t, 3H, J = 7.2Hz), 1.01 (t, 3H, J = 7.6Hz) ppm

[0364] LCMS:

[0365] Chromatographic column: Zorbax SB-C8 (2.1x 50mm; 1.8μm; RRHD 1200bar). Flow rate: 0.6ml / min, wavelength (λ): 215&246nm

[0366] Mobile phase A: 10 mM NH 4 OAc (water / methanol / acetonitrile 900 / 60 / 40)

[0367] Mobile phase B: 10 mM NH 4 OAc (water / methanol / acetonitrile 100 / 540 / 360)

[0368] Method: Mobile phase A / mobile phase B: 95 / 5 (0.0 min-2.0 min), 0 / 100 (1.5 min)

[0369] Results and conclusions. 100 g of PTS was synthesized with a yield of 65% and a purity of 97.6% (a / a), λ215nm and 98.7% (a / a), λ246nm.

[0370] Example 7. Dose-Response PTS Treatment and Milk Production

[0371] This example shows that dipropylthiosulfinate (PTS), a compound chemically related to DPD, exhibits the same or similar effects as DPD in reducing milk production. In addition, the correlation between different doses of PTS and reduced milk production is determined.

[0372] Study design. Twelve dairy cows (Holstein Friesian cows) from 1 farm were used. The dry period for each cow began 40 to 60 days before calving. Cows were milked twice a day at fixed times, and cows with a milk production of >12.5 L / day in the 24 hours before the administration of PTS were selected. The synthesis method of PTS is as described in Example 6. No additional measures were taken to stimulate dry milk, such as changing the feeding diet or changing the milking frequency. Cows were milked twice a day. Cows were housed in free stables. Porcine gelatin capsules containing PTS or placebo gel capsules containing water were administered 7 to 14 days before the start of the dry period.

[0373] In this study, different doses of synthetic PTS (purity >98%) were given to dairy cows with a daily milk production of at least 12.5 kg. The following doses were administered: 0, 10, 20, 40 ml of pure PTS. Two cows were used for each dose.

[0374] Milk production was monitored for 14 days before the different doses were given. The gel capsules were given orally with a pill booster and the gel capsules for the placebo group contained tap water. The capsules were given between morning and afternoon milking. At t=14 days, capsules containing PTS and placebo were given and milk production was monitored for a further 48 hours.

[0375] Results. Table 16 shows the milk production and the change in milk production (%) after administration of different doses of PTS. A positive value of "percent reduction" indicates a decrease in milk production, while a negative value indicates an increase in production. The reference treatment of administering water capsules instead of PTS did not show any effect after 24 and 48 hours. However, a dose of 20 ml of PTS significantly reduced milk production compared to the milk production before treatment, and the reducing effect became stronger when more PTS was administered.

[0376] Table 15. Milk production on days before and after administration of gel capsules containing PTS. Gel capsules were administered after the last milking at t=0.

[0377]

[0378] *: At t=0 day, a gel capsule containing PTS was administered.

[0379] LN = lactation number

[0380] PD = PTS dose (ml)

[0381] Table 16. Milk production and changes in milk production (expressed as percentage change compared to pre-treatment milk production) after administration of different doses of pure PTS.

[0382]

[0383]

[0384] Discussion and conclusions. This study showed that there was a dose-response curve between the dose of PTS and the decrease in daily milk production. The effect was evident after administration of 40 and 60 ml of PTS, but it cannot be ruled out that lower doses would also lead to a decrease in milk production. Higher doses may further reduce milk production.

[0385] It is clear from this study that other organosulfur analogs of DPD can also be used to achieve a temporary reduction in milk production, which allows cows more time to reduce their inherent milk production through other measures, such as by adjusting their diet. Because the reduction in milk production is temporary and recovers within 3-4 days of administration, this study also shows that PTS does not reduce milk production by damaging the udder. These treatments offer the possibility of preventing or reducing milk leakage and thus preventing new mastitis infections, and secondly, of reducing or preventing painful udder pressures associated with stress during the dry-off period. Therefore, giving cows PTS during the dry-off period is a valuable tool that benefits the health of the cow and the profitability of the farm.

[0386] Example 8. DPD analog treatment and milk production

[0387] In this example, it is demonstrated that treatment with chemically related compounds of DPD has the same or similar effect on the reduction of milk production as DPD. In this example, analogs as shown in Table 17 were used.

[0388] Study design. Thirty-six dairy cows (Holstein Friesian) from two farms were used. The cows were fed a diet containing TMR, including corn and grass silage, supplemented with soy and minerals.

[0389] The dry period for each cow started 40 to 60 days before calving. Cows were milked using a milking robot. Cows with a milk production >12.5 L / day in the 24 h before PTSO administration were selected. No additional measures were taken to stimulate dryness, such as changing the feeding diet or changing the milking frequency. Cows were milked twice a day. Cows were housed in free stables. Porcine gelatin capsules containing the designated compound or placebo gelatin capsules containing water were administered 7 to 14 days before the start of the dry period. Placebo treatment was equivalent to taking two gelatin capsules containing 180 ml of water.

[0390] In this study, the synthetic compounds shown in Table 17 (purity>96%) were administered to dairy cows. The purpose of this study was to compare the activity of the corresponding doses of the compounds in moles, based on the moles in 80 ml of di-n-propyl disulfide at 20°C. The moles were converted to milliliters or grams of the test compound. For this purpose, the molecular weight and specific density were used in the calculations. Table 17 lists the administered doses of the pure organosulfur analogs. As shown in the previous examples, the compounds were formulated in gelatin capsules.

[0391] Milk production was monitored for at least 5 days before the different doses were given. The gel capsules were given orally with a pill booster and the gel capsules for the placebo group contained tap water. The capsules were given between morning and afternoon milking. At t=0 day, the capsules and placebo capsules were given and milk production was monitored for 96 hours.

[0392] Results. Table 17 shows the relative changes (%) in milk production and milk yield. Positive values ​​for "percent reduction" indicate a decrease in milk production, while negative values ​​indicate an increase in production. A reference treatment was performed in which water capsules were given instead of the compound. A reduction in milk production was observed to varying degrees for all the tested compounds. Disulfides showed a significant effect, as did dialkyl (mono) sulfides, dialkyl trisulfides, dialkyl thiosulfinates, dialkyl thiosulfonates, and dialkyl sulfones.

[0393] The reduction in milk production was temporary: after 96 hours, milk production had almost fully recovered. This suggests that the reduction in milk production by administering these compounds is reversible and does not cause irreversible damage to the breast.

[0394] A further observation from these experiments was related to the odor during treatment. It was observed that there was a relationship between the degree of odor produced and the number of sulfur atoms in the treatment compound. For example, di-n-propyl trisulfide produced a greater odor than dipropyl disulfide, which in turn produced a greater odor than dipropyl monosulfide. Therefore, compounds with a monosulfide group were preferred.

[0395] These treatments offer the potential to prevent or reduce milk leakage, thereby preventing new mastitis infections, and secondly, to reduce or prevent painful udder pressure associated with stress during dry-off. Thus, administering these compounds to cows during dry-off is a valuable tool that benefits cow health and farm profitability.

Claims

1. Compounds of formula II: Formula II, wherein R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl; Q 1 Selected from the group consisting of -SS-, -S-, -SSS-, -S(O)2-, -S(O)-S- and -S(O)2-S-; Provided that the compound of formula II is not diphenyl disulfide; in, Preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups; Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl; Preferably, the C 1-4 Alkyl and benzyl groups are unsubstituted; or a composition comprising the compound, for d) reducing lactation in mammals; e) prophylactically treating intramammary infection in a mammal, and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; and / or f) Promote the health and welfare of lactating mammals.

2. Compounds of Formula I Formula I, Where R 1 and R 2 Independently selected from the group consisting of: C 1-4 Alkyl and benzyl; wherein, Preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted, or a combination of the compounds described herein is used d) reducing lactation in mammals; e) prophylactically treating intramammary infection in a mammal and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; and / or f) Promote the health and welfare of lactating mammals.

3. The compound or composition for use according to claim 1 or 2, wherein the mammal is a ruminant, preferably a cow.

4. A compound or composition for use as claimed in any one of the preceding claims, wherein R 1 and R 2 same.

5. A compound or composition for use according to any one of the preceding claims, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide and dibenzyl disulfide.

6. A compound or composition for use as claimed in any preceding claim, wherein the compound is di-n-propyl disulfide.

7. A compound or composition for use as claimed in any one of claims 1 or 3 to 4, wherein the compound is selected from di-n-propyl sulfide, diethyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfonate (PTSO) and di-n-propyl thiosulfinate (PTS).

8. A compound or composition for use as claimed in any preceding claim, wherein the use is combined with dry cow therapy.

9. A compound or composition for use according to any preceding claim, further comprising administering a prolactin inhibitor, such as cabergoline, quinagolide; casein hydrolysate; or an acid-producing mineral bolus.

10. A compound or composition for use according to any preceding claim, wherein the use further comprises the administration of an antibiotic, antifungal or anti-inflammatory agent.

11. A method for reducing lactation in a mammal; prophylactically treating intramammary infection in a mammal and / or reducing the occurrence of dry-off-related stress, dry-off-related inflammation or dry-off-related infection in a mammal; or promoting the health and welfare of a lactating mammal, comprising administering to a lactating mammal a pharmaceutical or veterinary composition or a functional food composition comprising a compound of formula II Formula II, wherein R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl; Q 1 Selected from the group consisting of -SS-, -S-, -SSS-, -S(O)2-, -S(O)-S- and -S(O)2-S-; Provided that the compound of formula II is not diphenyl disulfide; in, Preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups; Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl; Preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

12. The method of claim 11, wherein the compound is a compound of formula I Formula I, Where R 1 and R 2 Independently selected from the group consisting of: C 1-4 Alkyl and benzyl; wherein, Preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

13. A compound or composition for the method or use according to any one of the preceding claims, wherein the compound is formulated as a single dosage unit comprising at least 50 grams, preferably at least 70 grams of the compound, and the composition is administered to a pregnant ruminant.

14. A compound or composition for use according to any preceding claim, comprising selecting a pregnant cow producing at least 10 litres of milk per day and administering the composition to the cow.

15. A compound or composition for use according to any one of the preceding claims, wherein the mammal is abruptly or gradually stopped from milking and the compound is administered prior to or on the day of cessation of milking.

16. A pharmaceutical or veterinary composition or a functional food composition comprising a compound of formula II Formula II, wherein R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl, phenyl and benzyl; Q 1 Selected from the group consisting of -SS-, -S-, -SSS-, -S(O)2-, -S(O)-S- and -S(O)2-S-; Provided that the compound of formula II is not diphenyl disulfide; in, Preferably R 1 and R 2 Independently selected from the following group: C 1-4 Alkyl and benzyl groups; Preferably, the C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl or n-butyl; Preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

17. The composition of claim 16, wherein the compound is a compound of formula I Formula I, Where R 1 and R 2 Independently selected from the group consisting of: C 1-4 Alkyl and benzyl; wherein, Preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl or n-butyl; wherein, preferably, the C 1-4 The alkyl and benzyl groups are unsubstituted.

18. The composition of claim 16 or 17, wherein the compound is formulated as a single dosage unit comprising at least 50 grams of the compound, preferably at least 70 grams of the compound.

19. A composition according to any one of claims 16 to 18, wherein R 1 and R 2 same.

20. The composition of claim 19, wherein the compound is selected from di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, diisopropyl disulfide, di-n-butyl disulfide, dibenzyl disulfide, diethyl sulfide, di-n-propyl sulfide, diisopropyl sulfide, di-n-butyl sulfide, diphenyl sulfide, dibenzyl sulfide, di-n-propyl trisulfide, di-n-propyl sulfone, dibenzyl thiosulfinate, dibenzyl thiosulfinate, diisopropyl thiosulfinate, di-n-propyl thiosulfate (PTSO) and di-n-propyl thiosulfinate (PTS).

21. The composition of claim 19, wherein the compound is di-n-propyl disulfide.

22. A composition as claimed in any one of claims 16 to 21, further comprising a dry milk agent, preferably selected from a prolactin inhibitor, such as cabergoline, quinagolide; casein hydrolysate; or an acid-producing mineral bolus.

23. The composition of any one of claims 16-22, wherein the composition is a gel capsule.

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