CN114302647A - Approaches to ear protection against platinum-based antineoplastic agents - Google Patents

Abstract

Disclosed herein are methods of otoprotection against platinum-based anti-tumor agents by administering thiosulfate to a subject in need thereof. Typically, the thiosulfate salt is administered to the subject who is scheduled to administer a platinum-based anti-neoplastic agent within 4 hours. Alternatively, the thiosulfate salt is administered within 7 hours after administration of the platinum-based neoplastic agent.

Description

Approaches to ear protection against platinum-based antineoplastic agents

technical field

The present invention provides methods of ear protection against platinum-based antineoplastic agents.

Background technique

Platinum-based antineoplastic agents (eg, cisplatin) are chemotherapeutic agents widely used in the treatment of cancer and tumors. These agents are toxic and are known to induce hearing loss in both human and animal models. Thus, patients undergoing chemotherapy with platinum-based antineoplastic agents can suffer from hearing loss. There is a need for otoprotective compositions and methods to prevent or reduce hearing loss associated with chemotherapy regimens including platinum-based antineoplastic agents.

SUMMARY OF THE INVENTION

In general, the present invention provides methods for alleviating platinum-induced ototoxicity in a subject in need thereof. The method involves administering to a subject an effective amount of a thiosulfate.

In some embodiments, the subject is administered the platinum-based tumor agent no more than 7 hours prior to administration of the thiosulfate, or is scheduled to be administered the platinum-based anti-tumor agent within 4 hours. In certain embodiments, the subject is administered the platinum-based tumor agent no more than 7 hours prior to administration of the thiosulfate. In certain embodiments, the subject is scheduled to be administered a platinum-based antineoplastic agent at 4.5 hours. In additional embodiments, the subject is administered the platinum-based tumor agent no more than 2.5 hours prior to administration of the thiosulfate. In additional embodiments, the subject is administered the platinum-based tumor agent no more than 1 hour prior to administration of the thiosulfate.

In some embodiments, administering an effective amount of thiosulfate to a subject results in a plasma thiosulfate _Cmax of 30 μM or less upon administration of the platinum-based antineoplastic agent. In certain embodiments, administering an effective amount of thiosulfate to a subject produces a cochlear thiosulfate _Cmax that is at least 30 times greater than the cochlear _Cmax of the platinum-based antineoplastic agent. Cochlear platinum concentrations and cochlear _Cmax are typically modeled by pharmacokinetic simulations of intravenous infusion in a two-compartment model. For example, pharmacokinetic simulations can be performed using WinNonlin (Phoenix64) PK simulation model 9 (intravenous infusion, 2 compartment).

In additional embodiments, the thiosulfate is administered otically. In certain embodiments, the thiosulfate is administered intratympanically, transtympanically, or by inner ear injection. In certain embodiments, the thiosulfate is administered transtympanically or by inner ear injection.

In some embodiments, the method further comprises administering a platinum-based antineoplastic agent.

In certain embodiments, the thiosulfate salt is a basic thiosulfate salt, an ammonium thiosulfate salt, or a solvate thereof. In additional embodiments, the effective amount of thiosulfate is administered as a hypertonic pharmaceutical composition comprising an effective amount of thiosulfate. In additional embodiments, 200-1,000 μL (eg, 200-900 μL, 200-800 μL, 200-700 μL, 200-600 μL, 200-500 μL, 200-400 μL, 200-300 μL, 300-900 μL, 300-800 μL , 300-700 μL, 300-600 μL, 300-500 μL, 300-400 μL, 400-900 μL, 400-800 μL, 400-700 μL, 400-600 μL, or 400-500 μL) of the hypertonic pharmaceutical composition administered to the subject’s round window.

In additional embodiments, the calculated osmolality of the hypertonic pharmaceutical composition is 500-5,000 mOsm/L (eg, 600-5,000 mOsm/L, 700-5,000 mOsm/L, 800-5,000 mOsm/L, 900 -5,000mOsm/L, 1,000-5,000mOsm/L, 1,500-5,000mOsm/L, 2,000-5,000mOsm/L, 2,500-5,000mOsm/L, 3,000-5,000mOsm/L, 500-4,000mOsm/L, 600- 4,000mOsm/L, 700-4,000mOsm/L, 800-4,000mOsm/L, 900-4,000mOsm/L, 1,000-4,000mOsm/L, 1,500-4,000mOsm/L, 2,000-4,000mOsm/L, 2,500-4,000 mOsm/L, 3,000-4,000mOsm/L, 500-3,000mOsm/L, 600-3,000mOsm/L, 700-3,000mOsm/L, 800-3,000mOsm/L, 900-3,000mOsm/L, 1,000-3,000mOsm /L, 1,500-3,000mOsm/L, 2,000-3,000mOsm/L, 2,500-3,000mOsm/L, 500-2,500mOsm/L, 600-2,500mOsm/L, 700-2,500mOsm/L, 800-2,500mOsm/ L, 900-2,500mOsm/L, 1,000-2,500mOsm/L, 1,500-2,500mOsm/L, 2,000-2,500mOsm/L, 500-2,000mOsm/L, 600-2,000mOsm/L, 700-2,000mOsm/L , 800-2,000mOsm/L, 900-2,000mOsm/L, 1,000-2,000mOsm/L, 1,500-2,000mOsm/L, 500-1,500mOsm/L, 600-1,500mOsm/L, 700-1,500mOsm/L, 800-1,500mOsm/L, 900-1,500mOsm/L, or 1,000-1,500mOsm/L).

In some embodiments, the concentration of thiosulfate in the hypertonic pharmaceutical composition is 0.5M-2.5M (eg, about 0.05M to about 1.5M, about 0.05M to about 0.5M, about 0.05M to about 0.2M M, about 0.05M to about 0.1M, about 0.1M to about 1.5M, about 0.1M to about 0.5M, about 0.1M to about 0.2M, about 0.2M to about 1.5M, about 0.2M to about 0.5M, About 0.5M to about 1.5M, 0.05M to about 1.0M, about 0.05M to about 0.5M, about 0.05M to about 0.2M, about 0.05M to about 0.1M, about 0.1M to about 1.0M, about 0.1M to about 0.5M, about 0.1M to about 0.2M, about 0.2M to about 1.0M, about 0.2M to about 0.5M, about 0.5M to about 1.0M, or about 1.0M to about 1.5M).

In certain embodiments, an effective amount is an amount that results in a plasma thiosulfate concentration of 30 μM or less upon administration of the platinum-based antineoplastic agent. In certain embodiments, the effective amount is 0.1-2.5 mmol of thiosulfate. In particular embodiments, an effective amount is that amount that produces a maximum thiosulfate concentration of 0.6-10 mmol/L 1 h after administration. In further embodiments, an effective amount is an amount that produces a thiosulfate concentration of 0.1-2 mmol/L 7 h after administration in the cochlea of the subject.

In some embodiments, the subject is scheduled within about 1 hour to about 6 hours (eg, about 1 hour to about 5 hours, about 1 hour to about 4 hours, about 1 hour to about 1 hour after administration of the thiosulfate salt) About 3 hours, about 1 hour to about 2 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 2 hours to about 5 hours, about 2 hours to about 6 hours, about 3 hours to about 4 hours hours, about 3 hours to about 5 hours, about 3 hours to about 6 hours, about 4 hours to about 5 hours, about 4 hours to about 6 hours, or about 5 hours to about 6 hours) administration of the platinum-based anti-tumor agent.

In some embodiments, the subject is within about 1 hour to about 6 hours after administration of thiosulfate (eg, within about 1 hour to about 5 hours, about 1 hour to about 4 hours, About 1 hour to about 3 hours, about 1 hour to about 2 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 2 hours to about 5 hours, about 2 hours to about 6 hours, about 3 hours hours to about 4 hours, about 3 hours to about 5 hours, about 3 hours to about 6 hours, about 4 hours to about 5 hours, about 4 hours to about 6 hours, or about 5 hours to about 6 hours) administration based on Platinum antineoplastic agents.

In some embodiments, the subject is administered within about 1 hour to about 6 hours prior to administration of thiosulfate (eg, from about 1 hour to about 5 hours, about 1 hour to about 4 hours prior to administration of thiosulfate, About 1 hour to about 3 hours, about 1 hour to about 2 hours, about 2 hours to about 3 hours, about 2 hours to about 4 hours, about 2 hours to about 5 hours, about 2 hours to about 6 hours, about 3 hours hours to about 4 hours, about 3 hours to about 5 hours, about 3 hours to about 6 hours, about 4 hours to about 5 hours, about 4 hours to about 6 hours, or about 5 hours to about 6 hours) administration based on Platinum antineoplastic agents.

In additional embodiments, the invention is described by the items listed below.

1. A method of reducing platinum-induced ototoxicity in a subject in need, the method comprising administering to the subject an effective amount of thiosulfate, wherein the subject is prior to administering the thiosulfate The platinum-based neoplastic agent was administered no more than 7 hours, or the platinum-based antineoplastic agent was scheduled to be administered within 4 hours.

2. The method according to item 1, wherein the effective amount is an amount that results in a plasma thiosulfate concentration of 30 μM or less upon administration of the platinum-based antineoplastic agent.

3. A method of reducing platinum-induced ototoxicity in a subject in need thereof, the method comprising administering to the subject an effective amount of thiosulfate to produce (i) a platinum-based antineoplastic agent upon administration a plasma thiosulfate _Cmax of 30 μM or less, and (ii) a cochlear thiosulfate _Cmax that is at least 30 times greater than the cochlear _Cmax of the platinum-based antineoplastic agent, wherein the cochlear platinum concentration and This cochlear _Cmax was modeled by pharmacokinetic simulation of intravenous infusion in a two-compartment model.

4. The method of item 2, wherein the subject is administered a platinum-based tumor agent no more than 7 hours prior to administration of thiosulfate, or is scheduled to be administered a platinum-based anti-tumor agent within 4 hours.

5. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 7 hours prior to administration of the thiosulfate.

6. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 6 hours prior to administration of the thiosulfate.

7. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 5 hours prior to administration of the thiosulfate.

8. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 4 hours prior to administration of the thiosulfate.

9. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 3 hours prior to administration of the thiosulfate.

10. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 2.5 hours prior to administration of the thiosulfate.

11. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 2 hours prior to administration of the thiosulfate.

12. The method of any one of items 1 to 3, wherein the subject is administered the platinum-based tumor agent no more than 1 hour prior to administration of the thiosulfate.

13. The method of any one of items 1 to 3, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 4.5 hours.

14. The method of any one of items 1 to 3, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 4 hours.

15. The method of any one of items 1 to 3, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 3 hours.

16. The method of any one of items 1 to 3, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 2 hours.

17. The method of any one of items 1 to 3, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 1 hour.

18. The method according to any one of items 1 to 16, wherein the thiosulfate is administered otically.

19. The method of item 17, wherein the thiosulfate is administered intratympanically.

20. The method of item 17, wherein the thiosulfate is administered transtympanically.

21. The method according to item 17, wherein the thiosulfate is administered by inner ear injection.

22. The method of any one of items 1 to 20, further comprising administering a platinum-based antineoplastic agent.

23. The method of any one of items 1 to 21, wherein the thiosulfate salt is a basic thiosulfate salt, an ammonium thiosulfate salt, or a solvate thereof.

24. The method of any one of items 1 to 22, wherein the effective amount of thiosulfate is administered as a hypertonic pharmaceutical composition comprising an effective amount of thiosulfate.

25. The method according to item 23, wherein 200-1,000 μL of the hypertonic pharmaceutical composition is administered to the round window of the subject.

26. The method according to item 23 or 24, wherein the calculated osmolality of the hypertonic pharmaceutical composition is 500-5,000 mOsm/L.

27. The method according to any one of items 23 to 25, wherein the concentration of thiosulfate in the hypertonic pharmaceutical composition is 0.5M-2.5M.

28. The method according to any one of items 23 to 25, wherein the concentration of thiosulfate in the hypertonic pharmaceutical composition is 0.5M-1.5M.

29. The method according to any one of items 23 to 25, wherein the concentration of thiosulfate in the hypertonic pharmaceutical composition is 0.5M-1.0M.

30. The method of any one of items 1 to 28, wherein the effective amount is at least 0.05 mmol of thiosulfate.

31. The method of any one of items 1 to 28, wherein the effective amount is at least 0.1 mmol of thiosulfate.

32. The method of any one of items 1 to 28, wherein the effective amount is at least 0.2 mmol of thiosulfate.

33. The method of any one of items 1 to 28, wherein the effective amount is at least 0.3 mmol of thiosulfate.

34. The method of any one of items 1 to 28, wherein the effective amount is at least 0.4 mmol of thiosulfate.

35. The method of any one of items 1 to 33, wherein the effective amount is 2.5 mmol or less of thiosulfate.

36. The method of any one of items 1 to 33, wherein the effective amount is 2.0 mmol or less of thiosulfate.

37. The method of any one of items 1 to 33, wherein the effective amount is 1.5 mmol or less of thiosulfate.

38. The method of any one of items 1 to 33, wherein the effective amount is 1.0 mmol or less of thiosulfate.

39. The method of any one of items 1 to 33, wherein the effective amount is 0.5 mmol or less of thiosulfate.

40. The method according to any one of items 1 to 38, wherein the effective amount is the amount that produces a maximum thiosulfate concentration of 0.6-10 mmol/L 1 h after application.

41. The method according to any one of items 1 to 39, wherein the effective amount is an amount that produces a thiosulfate concentration of 0.1-2 mmol/L 7 h after administration in the cochlea of the subject.

definition

The term "about" as used herein means a value within ±10% of the value following the term "about".

The term "basic salt" as used herein represents a sodium or potassium salt of a compound. Basic salts can be monobasic, or binary or ternary if the number of acidic moieties (eg, -COOH, _-SO3H , or -P(O)(OH) _n moieties) allows.

The term "ammonium salt" as used herein represents the _NH4 ⁺ salt of a compound. Ammonium salts can be monobasic, or binary or ternary if the number of acidic moieties (eg, -COOH, _-SO3H , or -P(O)(OH) _n moieties) allows.

)、琼脂、明胶、葡甘露聚糖、半乳甘露聚糖(例如，瓜尔胶、槐豆胶或塔拉胶)、黄原胶、壳聚糖、果胶、淀粉、黄蓍胶、角叉菜胶、聚乙烯吡咯烷酮、聚乙烯醇、石蜡、凡士林、硅酸盐、丝蛋白，以及它们的组合。The term "gelling agent" as used herein refers to a pharmaceutically acceptable excipient known in the art for producing a gel when mixed with a solvent (eg, an aqueous solvent). Non-limiting examples of gelling agents include hyaluronic acid, polyoxyethylene-polyoxypropylene block copolymers (eg, poloxamers), poly(lactic acid-co-glycolic acid), polylactic acid, polycaprolactone , alginic acid or its salts, polyethylene glycol, cellulose, cellulose ethers, carbomers (for example,

), agar, gelatin, glucomannan, galactomannan (eg, guar gum, locust bean gum, or tara gum), xanthan gum, chitosan, pectin, starch, tragacanth, horn Carrageenan, polyvinylpyrrolidone, polyvinyl alcohol, paraffin, petrolatum, silicate, silk protein, and combinations thereof.

The term "hypertonic" as used herein in reference to a pharmaceutical composition means that the calculated osmolality of the pharmaceutical composition is from 300 mOsm/L to 7,000 mOsm/L (eg, 300 mOsm/L to 2,500 mOsm/L), the calculated molar The osmolarity corresponds to 300 mmol to 7,000 mmol (eg, 300 mOsm/L to 2,500 mmol) of ions and/or neutral molecules obtained by combining the platinum inactivator and any ionic non-polymeric excipients. The form was dissolved in 1 L of solvent with a calculated osmolality of 0 mOsm/L. For the purposes of this disclosure, calculated osmolality excludes ionic and/or neutral molecules generated by polymeric excipients (eg, by gelling agents). For the purposes of this disclosure, polymeric excipients (eg, gelling agents) are not considered to contribute to the calculated osmolality of the compositions disclosed herein.

The term "intratympanic" as used herein in relation to a route of administration refers to injection or infusion into a subject by injection or infusion through the ear canal with a temporarily removed or elevated tympanic membrane or through a port formed through an auditory bulla. into the middle ear and delivered to the round window.

The term "pharmaceutical composition" as used herein means a composition formulated with a pharmaceutically acceptable excipient and approved by a governmental regulatory agency for manufacture or sale as part of a therapeutic regimen for the treatment of a disease in a mammal.

The term "pharmaceutical dosage form" as used herein refers to those pharmaceutical compositions intended for administration to a subject without further modification (eg, without dilution, suspension or dissolution with a liquid solvent).

The term "pharmaceutically acceptable excipient" as used herein refers to any ingredient other than the thiosulfate and gelling agents described herein (eg, a vehicle capable of suspending or dissolving the active compound), and The ingredients have the properties of being substantially non-toxic to the patient and substantially non-inflammatory. Excipients may include, for example, antioxidants, disintegrants, dyes (colorants), softeners, emulsifiers, fillers (diluents), fragrances, fragrances, preservatives, printing inks, adsorbents, suspending agents or dispersions agents, sweeteners, liquid solvents and buffers.

The term "pharmaceutically acceptable salt" as used herein means suitable within the scope of sound medical judgment for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction, etc. and with a reasonable benefit/risk ratio commensurate with those salts. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66: 1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (edited by P.H. Stahl and C.G. Wermuth), Wiley-VCH, in 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting the free base group with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyric acid salt, camphorate, camphorsulfonate, citrate, cyclopentane propionate, digluconate, lauryl sulfate, ethanesulfonate, fumarate, glucose heptanoate ( glucoheptonate), glycerophosphate, hemisulfate, heptanoate, caproate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mesylate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate acid salt, pamoate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate , sulfate, tartrate, thiocyanate, tosylate, undecanoate, valerate, etc. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as nontoxic ammonium, quaternary ammonium and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine , dimethylamine, trimethylamine, triethylamine, ethylamine, etc.

The term "pharmaceutically acceptable solvate" as used herein refers to a compound as described herein wherein molecules of a suitable solvent are incorporated into a crystal lattice. Suitable solvents are physiologically tolerable at the doses administered. For example, solvates can be prepared by crystallization, recrystallization or precipitation from solutions comprising organic solvents, water, or mixtures thereof. Examples of suitable solvents are ethanol, water (eg, monohydrate, dihydrate, trihydrate, tetrahydrate, and pentahydrate), N-methylpyrrolidone (NMP), dimethylsulfoxide (DMSO) , N,N'-dimethylformamide (DMF), N,N'-dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU), 1,3- Dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone, benzyl benzoate, etc. . When the solvate is water-based, the solvate is referred to as a hydrate.

The term "platinum-based antineoplastic agent" as used herein represents a coordination compound of Pt(II) or Pt(IV). Platinum-based antineoplastic agents are known in the art as platins. Typically, platinum-based antineoplastic agents comprise at least two coordination sites at the platinum center occupied by one or more nitrogen-containing bystander ligands. Nitrogen-containing bystander ligands are monodentate or bidentate ligands in which the donor atom is a sp3- or ^{sp2 -} hybridized nitrogen atom within the ligand. Non-limiting examples of nitrogen-containing bystander ligands are ammonia, 1,2-cyclohexanediamine, picoline, phenanthrene, or 1,6-hexanediamine. Non-limiting examples of platinum-based antineoplastic agents include cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatinate tetranitrate, phenanthroplatin, picoplatin, and setteplatin.

The term "subject" as used herein refers to an animal (eg, a mammal, eg, a human). A subject treated according to the methods described herein can be a subject treated with a treatment regimen comprising a platinum-based antineoplastic agent (eg, a treatment regimen for the treatment of benign tumors, malignant tumors, or cancer). The subject may have been diagnosed with a benign tumor, malignant tumor or cancer by any method or technique known in the art. It will be understood by those of skill in the art that subjects to be treated in accordance with the present invention may have undergone standard testing, or may have been identified without examination as having undergone a treatment regimen including platinum-based antineoplastic agents. Subjects at high risk.

As used herein, the term "substantially neutral" refers to a pH level of 5.5 to about 8.5 as measured at 20°C.

The term "tonicity agent" as used herein refers to a class of pharmaceutically acceptable excipients used to control the osmolality of pharmaceutical compositions. Non-limiting examples of tonicity agents include substantially neutral buffers (eg, phosphate buffered saline, tris buffer, or artificial perilymph), dextrose, mannitol, glycerol, potassium chloride, and sodium chloride (eg, , hypertonic, isotonic, or hypotonic saline). Artificial perilymph contains NaCl (120-130 mM), KCl (3.5 mM), CaCl ₂ (1.3-1.5 mM), MgCl ₂ (1.2 mM), glucose (5.0-11 mM) and buffers (eg, NaHCO ₃ ( 25 mM) and _{NaH2PO4 (} 0.75 mM), or HEPES (20 mM) and NaOH (adjusted to a pH of about 7.5)) in water.

The term "transtympanic" as used herein in relation to a route of administration refers to delivery to the round window by transtympanic injection or infusion. Transtympanic injections can be performed directly through the tympanic membrane or through a tube embedded in the tympanic membrane (eg, via a tympanostomy tube or tympanic ventilation tube).

The term "inner ear injection" as used herein refers to the direct injection of a drug into the inner ear space.

Description of drawings

Figure 1 is a graph showing the profile of mean plasma thiosulfate concentration over time (0-24h) for each tested human cohort. The X-axis shows time (h) and the Y-axis shows mean plasma thiosulfate concentration (ng/mL).

Figure 2 is a graph showing the profile of mean plasma thiosulfate concentration over time (0-4h) for each tested human cohort. Error bars shown are standard deviation. The X-axis shows time (h) and the Y-axis shows mean plasma thiosulfate concentration (ng/mL).

Figure 3 is a graph showing the profile of mean plasma thiosulfate concentration over time (0-672h) for each tested human cohort. The X-axis shows time (h) and the Y-axis shows mean plasma thiosulfate concentration (ng/mL).

Figure 4 is a schematic diagram showing the timing of thiosulfate administration relative to cisplatin.

Figure 5A is a graph showing mean threshold sound pressure levels at 4 kHz, 24 kHz and 32 kHz measured during the Auditory Brainstem Response (ABR) test in control guinea pigs. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves. The X-axis shows the acoustic frequency in kHz, and the Y-axis shows the response threshold in decibels of sound pressure level (dB SPL).

Figure 5B is a graph showing the mean threshold sound pressure levels at 4 kHz, 24 kHz and 32 kHz measured during the auditory brainstem response (ABR) test of cisplatin challenged guinea pigs following administration of sodium thiosulfate to each ear . Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves. The X-axis shows the audio frequency in kHz, and the Y-axis shows the response threshold in decibels of sound pressure level (dBSPL). Data shown are mean ± standard error of the mean (SEM); two way analysis of variance (ANOVA); **P<0.01; ***P<0.001. Treated ears versus untreated ears (compare to Figure 5A).

Figure 6 is a graph showing the concentration dependence of thiosulfate (DB-020) in human tumor cell lines treated with 15 μM of cisplatin. The following tumor cell lines were used: SH-N-AS (brain, neuroblastoma), SNU899 (larynx, squamous cell carcinoma), NCI-H23 (lung, non-small cell), HLF (liver, undifferentiated liver) cell carcinoma) and A2780 (ovarian, malignant tumor).

Figure 7 is a graph showing the mean thiosulfate concentration (mM) in plasma (human) and perilymph (guinea pig) over time following administration of Hyaluronic Acid Gel 1 (12% w/v, 0.5 M sodium thiosulfate) A graph of the profile (0-8h) that goes over. The horizontal solid line shows the level of 30 μM, below which plasma thiosulfate levels should be. The horizontal dashed line shows the level of 660 [mu]M (0.66 mM), above which the perilymph thiosulfate level should be.

Figure 8A is a graph showing thiosulfate concentrations in plasma, perilymph, and cerebrospinal fluid over time in guinea pigs administered a gel containing 0.1 M sodium thiosulfate and 20% (w/v) poloxamer 407 chart.

Figure 8B is a graph showing changes in plasma, perilymph, and cerebrospinal fluid thiosulfate concentrations over time in guinea pigs administered a gel containing 0.5 M sodium thiosulfate and 1% (w/v) hyaluronic acid .

Figure 9A is a graph showing the change in thiosulfate concentration over time in plasma, perilymph and cerebrospinal fluid of guinea pigs administered a gel containing 0.1 M sodium thiosulfate and 2% (w/v) hyaluronic acid .

Figure 9B is a graph showing changes in plasma, perilymph and cerebrospinal fluid thiosulfate concentrations over time in guinea pigs administered a gel containing 0.5M sodium thiosulfate and 2% (w/v) hyaluronic acid .

Figure 10A is a graph showing threshold sound pressure levels at 4 kHz, 24 kHz and 32 kHz measured in five guinea pig cohorts (n=27 animals) during the auditory brainstem response test. All guinea pigs were injected intraperitoneally with cisplatin 7 days prior to auditory brainstem response testing. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves.

Figure 10B is a graph showing threshold sound pressure levels at 4 kHz, 24 kHz and 32 kHz measured in five guinea pig cohorts (n=18 animals) during the auditory brainstem response test. All guinea pigs were injected intraperitoneally with cisplatin 7 days prior to auditory brainstem response testing. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves.

Figure 11A is a graph showing mean threshold sound pressure levels at 4 kHz, 24 kHz and 32 kHz measured during auditory brainstem response testing of guinea pigs (n=18 animals) thought to have hearing loss. All guinea pigs were injected intraperitoneally with cisplatin 7 days prior to auditory brainstem response testing. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves.

Figure 11B is a graph showing the mean threshold sound at 4 kHz, 24 kHz and 32 kHz measured during the auditory brainstem response (ABR) test of cisplatin challenged guinea pigs following vehicle or sodium thiosulfate administration to each ear Pressure level chart. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves.

Figure 12 is a graph showing a cisplatin challenge test following administration of vehicle or sodium thiosulfate to one ear of a guinea pig.

Figure 13 is a graph showing the auditory brainstem responses of cisplatin challenged guinea pigs after vehicle or sodium thiosulfate (0.1 M, 0.5 M or 1 M sodium thiosulfate gel) was applied to each ear ( ABR) graph of the average threshold sound pressure levels at 4kHz, 24kHz and 32kHz measured during the test. Baseline thresholds were derived from historical auditory brainstem response tests on cisplatin-naive guinea pigs (n=100 ears). Baseline thresholds are shown as shaded area curves.

Detailed ways

In general, the present invention provides methods of alleviating platinum-induced ototoxicity in a subject by administering to the subject an effective amount of thiosulfate. Preferably, the thiosulfate is administered otically (eg, intratympanically or transtympanically).

Typically, thiosulfate is administered to a subject who is scheduled to be administered a platinum-based antineoplastic agent within 4 hours (eg, within 3 hours, within 2 hours, or within 1 hour). Alternatively, the thiosulfate salt is administered within 7 hours (eg, within 6 hours, within 5 hours, within 4 hours, within 3 hours, within 2 hours, or within 1 hour) of administration of the platinum-based tumor agent. Preferably, thiosulfate is administered to a subject who is scheduled to be administered a platinum-based antineoplastic agent within 3 hours. Alternatively, thiosulfate is administered within 4 hours of administration of the platinum-based tumor agent. More preferably, the thiosulfate is administered to a subject who is scheduled to be administered a platinum-based antineoplastic agent within 1 hour. Alternatively, thiosulfate is administered within 1 hour of administration of the platinum-based tumor agent.

An effective amount of thiosulfate typically produces plasma thiosulfate concentrations of 30 μM or less (eg, 20 μM or less, 10 μM or less, or near endogenous concentrations) upon administration of platinum-based antineoplastic agents . Additionally or alternatively, an effective amount of thiosulfate typically results in a cochlear _Cmax that is at least 30-fold greater (eg, 30- to 1000-fold greater, 30- to 500-fold greater, or 30- to 150-fold greater cochlear thiosulfate concentrations), where the cochlear platinum concentration and the cochlear _Cmax were modeled by pharmacokinetic simulations of intravenous infusion in a two-compartment model.

Platinum-induced ototoxicity can occur in subjects receiving platinum-based antineoplastic agents (eg, subjects with tumors or cancer). Non-limiting examples of platinum-based antineoplastic agents include cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatinate tetranitrate, phenanthroplatin, picoplatin, and setteplatin.

Thiosulfate can reduce (eg, eliminate) hearing loss in subjects receiving a platinum-based antineoplastic agent, such as by relative to those receiving the same platinum-based antineoplastic agent regimen but not receiving thiosulfate. With reference to the subject, at least 50% (eg, at least 60%, at least 70%, or at least 80% of the subject's sound pressure level threshold assessment) at frequencies of 8 kHz or higher (eg, between 8 kHz and 20 kHz) %) reduction measured.

Thiosulfate salts can exhibit otoprotective properties against platinum-based antineoplastic agents, and can be used in methods for alleviating (eg, eliminating) platinum-induced ototoxicity in a subject in need thereof. Typically, thiosulfate is administered to the subject's round window. The subject may be undergoing therapy with a platinum-based antineoplastic agent (eg, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatinate tetranitrate, phenanthroplatin, picoplatin, or setteplatin).

The thiosulfate salt can be administered to the subject, eg, before or after administration of the platinum-based antineoplastic agent to the subject. Alternatively, the thiosulfate salt can be administered, eg, at the same time as the platinum-based antineoplastic agent. Can be administered to a drug that is scheduled to be administered within 4 hours, eg, within 3 hours, within 2 hours, or within 1 hour (eg, at least 5 minutes, at least 15 minutes, or at least 30 minutes prior to administration of the platinum-based antineoplastic agent). Subjects with platinum antineoplastic agents were administered thiosulfate. Alternatively, the platinum-based antineoplastic agent may be administered (eg, at least 5 minutes, at least 15 minutes, or at least 30 minutes after administration), eg, no more than 7 hours (eg, no more than 6 hours, no more than 5 hours, no more than 7 hours) 4 hours, no more than 3 hours, no more than 2 hours, or no more than 1 hour) administration of thiosulfate. Administration of platinum-based antineoplastic agents typically precedes administration of a hydrating composition (eg, physiological saline optionally containing, eg, mannitol or furosemide). Furthermore, administration of the platinum-based antineoplastic agent typically continues for a period of at least 1 hour (eg, 1 to 24 hours, such as 1 to 12 hours, 1 to 6 hours, 1 to 3 hours, or 1 to 2 hours). Those skilled in the art will recognize that the timing between the administration of the thiosulfate and the platinum-based tumor agent is the time between the completion of administration of one and the initiation of administration of the other. For example, administration of a platinum-based tumor agent 30 minutes to 4 hours after the step of administering thiosulfate indicates the end of thiosulfate administration and the beginning of platinum-based tumor agent administration (eg, cisplatin 30 minutes to 4 hours) infusion) separately.

Generally, the pharmaceutical compositions of the present invention can be administered by routes other than platinum-based antineoplastic agents. The methods of the present invention may utilize a topical route of administration, eg, the pharmaceutical compositions of the present invention may be administered intratympanically or transtympanically. Transtympanic administration can include injecting or infusing an effective amount of a pharmaceutical composition of the invention through the tympanic membrane into the tympanic cavity, thereby providing the anti-platinum chemoprotectant to the round window.

In the method of the present invention, a needle is typically used to pierce the tympanic membrane to instill the drug into the middle ear space, or through an existing PE tube or perforation of the ear drum to instill the drug. A separate vent may or may not be created in the eardrum to allow air to escape the middle ear space. The instilled drug can then target middle ear structures, cells, or be designed to enter the inner ear through circular and oval membranes to affect specific targets. This can be achieved, for example, by instilling the drug through the round window membrane, oval window, cochleostomy or labrinthotomy. These surgical procedures can be accomplished by raising the tympanic canal flap (lifting the tympanic membrane) and exposing the round window, stapes/oval window, and promontory. A stapediotomy hole can be created in the footplate and the drug instilled into the vestibule by pump, injection, or some other method. Alternatively, the bony lip of the round window (RW) is removed (usually by drilling) to expose the RW. The RW can then be punctured with a needle and the drug infused, or the RW can be fenestrated and the drug infused directly through the fenestration. Finally, a completely separate cochlear entry hole can be opened by drilling a cochleostomy hole in the cochlea, and medication can be instilled.

Alternatively, instead of elevating the tympanic canal flap, a mastoidotomy can be performed and the facial recesses opened to provide direct access to the oval and round windows, as well as the promontory and semicircular canals. In this way, all three sites can be used as just described. In addition, the labyrinth can be opened for drug instillation much like a cochleostomy. To dissipate fluid/pressure buildup in the inner ear, separate openings into the RW or OW can be created to allow excess perilymph to leak.

Thiosulfate salts can be provided in pharmaceutical compositions. The pharmaceutical composition can be, for example, hypertonic. Without wishing to be bound by theory, the higher tonicity of the pharmaceutical compositions disclosed herein is believed to increase the thiolation at the round window of the subject relative to compositions with lower tonicity (eg, hypotonic or isotonic) Sulfate bioavailability. Bioavailability is typically calculated using thiosulfate exposure (AUC) following administration of thiosulfate to a subject. The calculated osmolality of the pharmaceutical composition (eg, pharmaceutical dosage form) can be, for example, at least 400 mOsm/L (eg, at least 500 mOsm/L, at least 600 mOsm/L, at least 700 mOsm/L, at least 800 mOsm/L, at least 900 mOsm/L, at least 1,000 mOsm/L, at least 1,500 mOsm/L, at least 2,000 mOsm/L, at least 2,500 mOsm/L, or at least 3,000 mOsm/L), and/or 5,000 mOsm/L or less (eg, 4,000 mOsm/L or Less, 3,000mOsm/L or less, 2,000mOsm/L or less, 1,900mOsm/L or less, 1,800mOsm/L or less, 1,700mOsm/L or less, 1,600mOsm/L or less , or 1,500mOsm/L or less). The calculated osmolality of a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, 1,500-4,500 mOsm/L. The calculated osmolality of a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, 3,000-4,500 mOsm/L. The measured osmolarity of a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, at least 0.3 Osm/kg (eg, at least 0.5 Osm/kg, at least 0.6 Osm/kg, at least 0.7 Osm/kg, at least 0.8 Osm/kg , at least 0.9 Osm/kg, at least 1.0 Osm/kg, at least 1.2 Osm/kg, at least 1.4 Osm/kg, or at least 1.8 Osm/kg). The measured osmolality of a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, 2.5 Osm/kg or less (eg, 2.1 Osm/kg or less). The measured osmolarity of a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, 0.3-2.5 Osm/kg (eg, 0.5-2.5 Osm/kg, 0.6-2.5 Osm/kg, 0.7-2.5 Osm/kg, 0.8 -2.5Osm/kg, 0.9-2.5Osm/kg, 1.0-2.5Osm/kg, 1.2-2.5Osm/kg, 1.4-2.5Osm/kg, 1.8-2.5Osm/kg, 0.5-2.1Osm/kg, 0.6- 2.1Osm/kg, 0.7-2.1Osm/kg, 0.8-2.1Osm/kg, 0.9-2.1Osm/kg, 1.0-2.1Osm/kg, 1.2-2.1Osm/kg, 1.4-2.1Osm/kg, or 1.8- 2.1Osm/kg). "Calculated osmolality" refers to the millimoles of ionic and/or neutral molecules produced by dissolving one or more compounds in 1 L of deionized or distilled water; calculated osmolality does not include those imparted by polymerization Ionic and/or neutral molecules produced by excipients (eg, from gelling agents). "Measured osmolality" refers to the osmolarity of a composition as measured using an osmometer (usually a membrane osmometer).

The preferred pharmaceutical dosage form of the present invention is a gel.

In some embodiments, at least 50 μL (preferably, at least 100 μL; more preferably, at least 200 μL) of the pharmaceutical composition is administered to the subject's round window. In certain embodiments, 1 mL or less (eg, 0.8 mL or less, or 0.5 mL or less) of the pharmaceutical composition is administered to the subject's round window. In certain embodiments, 100 μL to 1 mL (e.g., 200 μL to 1 mL, 100 μL to 0.8 mL, 200 μL to 0.8 mL, 100 μL to 0.5 mL, 200 μL to 0.5 mL, 0.5 mL to 1.0 mL, 0.5 mL to 0.8 mL, or 0.8 mL to 1.0 mL) of the pharmaceutical composition is administered to the round window of the subject.

Thiosulfate can be, for example, the only compound that contributes to the osmolality of the pharmaceutical composition. Alternatively, a higher osmolarity than that provided by the desired concentration of thiosulfate can be achieved, such as through the use of tonicity agents. The tonicity agent can be present in a hypertonic, isotonic or hypotonic vehicle (eg, a hypotonic liquid solvent). Non-limiting examples of tonicity agents include substantially neutral buffers (eg, phosphate buffered saline, tris buffer, or artificial perilymph), dextrose, mannitol, glycerin, glycerol, chlorine Potassium chloride and sodium chloride (eg, hypertonic, isotonic, or hypotonic saline).

Thiosulfate

Without wishing to be bound by theory, thiosulfates are believed to reduce or eliminate the toxicity of platinum-based antineoplastic agents by competitively linking and substantially coordinatively saturating platinum centers present in platinum-based antineoplastic agents. The concentration of thiosulfate in a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, at least about 0.05M (eg, at least about 0.1M, at least about 0.2M, at least about 0.3M, at least about 0.4M, at least about 0.5M) M, or at least about 1M). The concentration of thiosulfate in a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, about 2.5M or less (eg, 2.0M or less, 1.5M or less, 1.0M or less, 0.5M or lower, about 0.3M or lower, or about 0.2M or lower). Non-limiting examples of concentrations of thiosulfate in a pharmaceutical composition (eg, a pharmaceutical dosage form) can be, for example, about 0.05M to about 1.5M, about 0.05M to about 0.5M, about 0.05M to about 0.2M, about 0.05M M to about 0.1M, about 0.1M to about 1.5M, about 0.1M to about 0.5M, about 0.1M to about 0.2M, about 0.2M to about 1.5M, about 0.2M to about 0.5M, about 0.5M to about 1.5M, 0.05M to about 1.0M, about 0.05M to about 0.5M, about 0.05M to about 0.2M, about 0.05M to about 0.1M, about 0.1M to about 1.0M, about 0.1M to about 0.5M , about 0.1M to about 0.2M, about 0.2M to about 1.0M, about 0.2M to about 0.5M, about 0.5M to about 1.0M, or about 1.0M to about 1.5M. Preferably, the concentration of the thiosulfate agent in the pharmaceutical composition (eg, pharmaceutical dosage form) is from about 0.5M to about 1.5M. More preferably, the concentration of the thiosulfate agent in the pharmaceutical composition (eg, pharmaceutical dosage form) is from about 0.5M to about 1.0M.

Preferably, the thiosulfate is an alkaline thiosulfate or ammonium thiosulfate. More preferably, the thiosulfate salt is sodium thiosulfate.

gelling agent

The pharmaceutical compositions disclosed herein comprise a gelling agent. Gelling agents can be used to increase the viscosity of the pharmaceutical composition, thereby improving retention of the pharmaceutical composition at the target site. A pharmaceutical composition (eg, a pharmaceutical dosage form) can contain, for example, from about 0.1% to about 25% (w/v) (eg, from about 0.1% to about 20% (w/v), from about 0.1% to about 10%) relative to the solvent % (w/v), about 0.1% to about 2% (w/v), about 0.5% to about 25% (w/v), about 0.5% to about 20% (w/v), about 0.5% to about 10% (w/v), about 0.5% to about 2% (w/v), about 1% to about 20% (w/v), about 1% to about 10% (w/v), about 1 % to about 2% (w/v), about 5% to about 20% (w/v), about 5% to about 10% (w/v), or about 7% to about 10% (w/v) ) gelling agent. Preferably, the pharmaceutical composition (eg, pharmaceutical dosage form) may contain, for example, about 0.5% to about 25% (w/v) (eg, about 0.5% to about 20% (w/v), about 0.5%) relative to the solvent to about 10% (w/v), about 0.5% to about 2% (w/v), about 1% to about 20% (w/v), about 1% to about 10% (w/v), about 1% to about 2% (w/v), about 5% to about 20% (w/v), about 5% to about 10% (w/v), or about 7% to about 10% (w/v) )) gelling agent.

)、琼脂、明胶、葡甘露聚糖、半乳甘露聚糖(例如，瓜尔胶、槐豆胶或塔拉胶)、黄原胶、壳聚糖、果胶、淀粉、黄蓍胶、角叉菜胶、聚乙烯吡咯烷酮、聚乙烯醇、石蜡、凡士林、硅酸盐、丝蛋白，以及它们的组合。本文所述的胶凝剂是本领域中已知的。优选地，所述胶凝剂是透明质酸。Gelling agents that can be used in the pharmaceutical compositions disclosed herein are known in the art. Non-limiting examples of gelling agents include hyaluronic acid, polyoxyethylene-polyoxypropylene block copolymers (eg, poloxamers), poly(lactic acid-co-glycolic acid), polylactic acid, polycaprolactone , alginic acid or its salts, polyethylene glycol, cellulose, cellulose ethers, carbomers (for example,

), agar, gelatin, glucomannan, galactomannan (eg, guar gum, locust bean gum, or tara gum), xanthan gum, chitosan, pectin, starch, tragacanth, horn Carrageenan, polyvinylpyrrolidone, polyvinyl alcohol, paraffin, petrolatum, silicate, silk protein, and combinations thereof. The gelling agents described herein are known in the art. Preferably, the gelling agent is hyaluronic acid.

The pharmaceutical composition may contain, for example, from about 0.5% to about 2% (w/v) (eg, from about 1% to about 2% (w/v)) of hyaluronic acid relative to the solvent. The pharmaceutical composition may contain, for example, about 5% to about 10% (w/v) (eg, about 6% to about 8% (w/v)) methylcellulose relative to the solvent. The pharmaceutical composition may contain, for example, hyaluronic acid and methylcellulose as gelling agents (eg, from about 0.5% to about 2% (w/v) of hyaluronic acid and from about 5% to about 10% (w/v) of hyaluronic acid relative to the solvent. /v) of methylcellulose). Pharmaceutical compositions may contain, for example, polyoxyethylene-polyoxypropylene block copolymers (eg, poloxamers) as gelling agents. The pharmaceutical composition may contain, for example, from about 1% to about 20% (w/v) (eg, from about 1% to about 15% (w/v), from about 1% to about 10% (w/v), relative to the solvent) , about 5% to about 20% (w/v), about 5% to about 15% (w/v), about 5% to about 10% (w/v), about 10% to about 20% (w/v) v), or from about 10% to about 15% (w/v)) of a polyoxyethylene-polyoxypropylene block copolymer (eg, a poloxamer). The poloxamer can be Poloxamer 407, Poloxamer 188, or a combination thereof. The pharmaceutical composition may contain, for example, from about 0.5% (w/v) to about 20% (w/v) of silk protein as a gelling agent, relative to the solvent.

Hyaluronan is hyaluronic acid or a salt thereof (eg, sodium hyaluronate). Hyaluronan is known in the art and is commonly obtained from various bacteria (eg, Streptococcus zooepidemicus, Streptococcus equi, or Streptococcus pyrogenes) or isolated from other sources (eg, bovine vitreous humor or cockscomb). Hyaluronic acid typically has a weight average molecular weight ( _Mw ) of from about 50 kDa to about 10 MDa. Preferably, the MW of hyaluronic acid (eg, sodium hyaluronate) is about 500 kDa to 6 _MDa (eg, about 500 kDa to about 750 kDa, about 600 kDa to about 1.1 MDa, about 750 kDa to about 1 MDa, about 1 MDa to about 1.25 MDa) , about 1.25 to about 1.5 MDa, about 1.5 MDa to about 1.75 MDa, about 1.75 MDa to about 2 MDa, about 2 MDa to about 2.2 MDa, about 2 MDa to about 2.4 MDa). More preferably, the MW of hyaluronic acid (eg, sodium hyaluronate) is about 620 kDa to about 1.2 MDa or about 1.2 _MDa to about 1.9 MDa. Other preferred molecular weight ranges for hyaluronic acid include, for example, from about 600 kDa to about 1.2 MDa.

和

商购。药物组合物可含有例如聚氧乙烯-聚氧丙烯嵌段共聚物(例如，泊洛沙姆)，所述共聚物包含数均分子量(M_n)为例如约1,100g/mol至约17,400g/mol(例如，约2,090g/mol至约2,360g/mol、约7,680g/mol至约9,510g/mol、6,830g/mol至约8,830g/mol、约9,840g/mol至约14,600g/mol、或约12,700g/mol至约17,400g/mol)的聚氧丙烯嵌段。聚氧乙烯-聚氧丙烯嵌段共聚物(例如，泊洛沙姆)可包含数均分子量(M_n)为约1,100g/mol至约4,000g/mol的聚氧丙烯嵌段和为约30％至约85％(w/w)的计算聚氧乙烯含量。优选地，聚氧乙烯-聚氧丙烯嵌段共聚物(例如，泊洛沙姆)可包含计算分子量为例如约1,800g/mol至约4,000g/mol的聚氧丙烯嵌段。优选地，聚氧乙烯-聚氧丙烯嵌段共聚物(例如，泊洛沙姆)的计算聚氧乙烯含量可为例如约70％至约80％(w/w)。优选地，聚氧乙烯-聚氧丙烯嵌段共聚物(例如，泊洛沙姆)的数均分子量可为例如约7,680g/mol至约14,600g/mol。泊洛沙姆的非限制性示例是泊洛沙姆407和泊洛沙姆188。Polyoxyethylene-polyoxypropylene block copolymers are known in the art. A non-limiting example of a polyoxyethylene-polyoxypropylene block copolymer is a poloxamer in which a single polyoxypropylene block is flanked by two polyoxyethylene blocks. Poloxamers are known under various trade names such as

and

Commercially available. The pharmaceutical composition may contain, for example, a polyoxyethylene-polyoxypropylene block copolymer (eg, a poloxamer) comprising a number average molecular weight (M _n ) of, for example, about 1,100 g/mol to about 17,400 g/mol mol (eg, about 2,090 g/mol to about 2,360 g/mol, about 7,680 g/mol to about 9,510 g/mol, 6,830 g/mol to about 8,830 g/mol, about 9,840 g/mol to about 14,600 g/mol , or about 12,700 g/mol to about 17,400 g/mol) of polyoxypropylene blocks. The polyoxyethylene-polyoxypropylene block copolymer (eg, poloxamer) may comprise polyoxypropylene blocks having a number average molecular weight ( _Mn ) of about 1,100 g/mol to about 4,000 g/mol and a polyoxypropylene block of about 30 % to about 85% (w/w) calculated polyoxyethylene content. Preferably, the polyoxyethylene-polyoxypropylene block copolymer (eg, poloxamer) may comprise a polyoxypropylene block having a calculated molecular weight of, for example, from about 1,800 g/mol to about 4,000 g/mol. Preferably, the calculated polyoxyethylene content of the polyoxyethylene-polyoxypropylene block copolymer (eg, poloxamer) may be, for example, from about 70% to about 80% (w/w). Preferably, the number average molecular weight of the polyoxyethylene-polyoxypropylene block copolymer (eg, poloxamer) may be, for example, from about 7,680 g/mol to about 14,600 g/mol. Non-limiting examples of poloxamers are Poloxamer 407 and Poloxamer 188.

Methocel^TM、

和

商购。纤维素醚的非限制性示例包括甲基纤维素、羧甲基纤维素、乙基纤维素、羟乙基纤维素、甲基羟乙基纤维素、羟丙基甲基纤维素或羟丙基纤维素。纤维素醚(例如，甲基纤维素)的数均分子量(M_n)可为例如约5kDa至约300kDa。甲基取代的纤维素(例如，甲基纤维素、羟丙基甲基纤维素或甲基羟乙基纤维素)可以具有例如19％至35％(例如，19％至30％)的甲基含量。Cellulose and cellulose ethers are known in the art. Cellulose and cellulose ethers are various trade names such as

Methocel ^TM ,

and

Commercially available. Non-limiting examples of cellulose ethers include methyl cellulose, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methyl hydroxyethyl cellulose, hydroxypropyl methyl cellulose, or hydroxypropyl cellulose cellulose. The number average molecular weight (M _n ) of the cellulose ether (eg, methylcellulose) can be, for example, from about 5 kDa to about 300 kDa. Methyl-substituted cellulose (eg, methylcellulose, hydroxypropylmethylcellulose, or methylhydroxyethylcellulose) can have, for example, 19% to 35% (eg, 19% to 30%) methyl content.

Silk protein is a protein present in silk produced by many insects. Silk proteins are known in the art and are commercially available from various suppliers such as Jiangsu SOHO International Group; Simatech, Suzhou, China; Xi'an Lyphar Biotech, Ltd.; Xi'an Rongsheng Biotechnology; Mulberry Farms, Treenway Silks, Sharda Group, Maniar Enterprises and Wild Fibres. The molecular weight of silk proteins is generally from about 10 kDa to about 500 kDa. Silk proteins are described in WO 2017/139684, the disclosure of which is incorporated herein by reference.

cross-linked gelling agent

The pharmaceutical composition may contain non-crosslinked or crosslinked gelling agents. The gelling agent can be cross-linked using cross-linking agents known in the art. Preferably, the cross-linked gelling agent is covalently cross-linked. Pharmaceutical compositions (eg, pharmaceutical dosage forms) comprising cross-linked gelling agents can be used to control the release profile of anti-platinum chemoprotectants. For example, the release of an anti-platinum chemoprotectant from a pharmaceutical composition (eg, a pharmaceutical dosage form) containing a cross-linked gelling agent may be prolonged relative to a reference composition that differs from the pharmaceutical composition The only difference is the lack of cross-linking of the gelling agent in the reference composition. Prolonged release of anti-platinum chemoprotectants can be assessed by comparing the _Tmax values of the pharmaceutical and reference compositions.

Certain gelling agents, such as those having carboxylate moieties (eg, hyaluronic acid, alginic acid, and carboxymethyl cellulose), may use ionic crosslinkers (eg, polyvalent metal ^ions such as Mg2 ⁺ , Ca ²⁺ or Al ³⁺ ) for ionic crosslinking. Ionic crosslinking techniques for gelling agents are known in the art (see, eg, US Pat. Nos. 6,497,902 and 7,790,699, the disclosures of which are incorporated herein by reference). Typically, the gelling agent can be ionically crosslinked in an aqueous solution using a multivalent metal ion (eg, ^Mg2+ , Ca2 ⁺ , or Al3 ⁺ ) as an ionic crosslinking agent. Without wishing to be bound by theory, it is believed that the metal ions coordinate with different molecules of the gelling agent (eg, with pendant carboxylic acid groups located on different molecules of the gelling agent), thereby forming between these different molecules of the gelling agent Connect key.

Certain gelling agents with reactive functional groups (eg, -OH, -COOH, or _-NH2 ) can be covalently cross-linked. Gelling agent covalent cross-linking techniques are known in the art (see, eg, Khunmanee et al., J. Tissue Eng., 8:2041731417726464, 2017, the disclosure of which is incorporated herein by reference). Non-limiting examples of covalent crosslinkers include: 1,4-butanediol diglycidyl ether (BDDE), divinyl sulfone, glutaraldehyde, cyanogen bromide, octyl succinic anhydride, acid chlorides, diisocyanates, Methacrylic anhydride, boric acid, and sodium periodate/adipic acid dihydrazide.

other excipients

The pharmaceutical compositions may contain pharmaceutical excipients other than gelling agents. For example, pharmaceutical compositions may contain, for example, liquid solvents, tonicity agents, buffers and/or colorants. Certain excipients can perform multiple roles. For example, in addition to its function as a carrier, liquid solvents can also act as tonicity and/or buffering agents. Such solvents are known in the art, such as saline (eg, hypertonic, hypotonic, isotonic, or phosphate buffered saline) and artificial perilymph.

Liquid solvents can be used as vehicles in pharmaceutical compositions (eg, pharmaceutical dosage forms). Liquid solvents are known in the art. Non-limiting examples of liquid solvents include water, saline (eg, hypertonic, hypotonic, isotonic, or phosphate buffered saline), artificial perilymph, and tris buffer. Artificial perilymph contains NaCl (120-130 mM), KCl (3.5 mM), CaCl ₂ (1.3-1.5 mM), MgCl ₂ (1.2 mM), glucose (5.0-11 mM) and buffers (eg, NaHCO ₃ ( 25 mM) and _{NaH2PO4 (} 0.75 mM), or HEPES (20 mM) and NaOH (adjusted to a pH of about 7.5)) in water.

Tonicity agents can be included in pharmaceutical compositions (eg, pharmaceutical dosage forms) to increase the osmolarity relative to that provided by the anti-platinum chemoprotectant. Tonicity agents are known in the art. Non-limiting examples of tonicity agents include substantially neutral buffers (eg, phosphate buffered saline, tris buffer, or artificial perilymph), dextrose, mannitol, glycerol, potassium chloride, and sodium chloride (eg, , hypertonic, isotonic, or hypotonic saline). The pharmaceutical composition (eg, pharmaceutical dosage form) comprises a sufficient amount of the tonicity agent to provide a hypertonic pharmaceutical dosage form for administration to a subject (eg, a calculated osmolality of at least 400 mOsm/L (eg, at least 500 mOsm/L) , at least 600mOsm/L, or at least 700mOsm/L), and/or 2,500mOsm/L or less (eg, 2,000mOsm/L, 1,900mOsm/L or less, 1,800mOsm/L or less, 1,700mOsm/L L or less, 1,600 mOsm/L or less, or 1,500 mOsm/L or less) pharmaceutical dosage form). For example, the target concentration of a tonicity agent in a pharmaceutical composition (eg, a pharmaceutical dosage form) can be determined, for example, by (i) subtracting anti-platinum chemoprotectants and other non-polymeric osmolarities from the total target osmolarity Calculated osmolality contribution of the excipient to obtain the target calculated osmolarity contribution from the tonicity agent, and (ii) by dividing the target calculated osmolarity contribution from the tonicity agent by when the tonicity agent The concentration of the tonicity agent is determined by the number of ions and/or molecules produced when dissolved in a liquid solvent. Accordingly, appropriate amounts of tonicity agents can be included in pharmaceutical compositions (eg, pharmaceutical dosage forms).

Buffers can be used to adjust the pH of a pharmaceutical composition (eg, a pharmaceutical dosage form) to a substantially neutral pH level. Buffers are known in the art. Non-limiting examples of buffers include, for example, phosphate buffers and Goode buffers (eg, tris, MES, MOPS, TES, HEPES, HEPPS, tris(hydroxymethyl)methylglycine (tricine) and N,N- bis(hydroxyethyl)glycine (bicine). In addition to pH control, buffers can be used to control the osmolarity of pharmaceutical compositions (eg, pharmaceutical dosage forms).

Preparation

Pharmaceutical compositions (eg, pharmaceutical dosage forms) of the present invention can be prepared from platinum-resistant chemoprotectants, gelling agents, and liquid solvents. The present invention provides a method of preparing a pharmaceutical composition (eg, a pharmaceutical dosage form) of the present invention by (i) providing an anti-platinum chemoprotectant and a gelling agent, and (ii) providing the anti-platinum chemoprotectant The gelling agent and the gelling agent are mixed with a liquid solvent to produce a hypertonic pharmaceutical composition.

The anti-platinum chemoprotectant and gelling agent can be provided, for example, as a mixture or as separate components. When the anti-platinum chemoprotectant and the gelling agent are provided separately, step (ii) may include, for example:

(a) first mixing a liquid solvent with a gelling agent to produce an intermediate mixture, and then mixing the intermediate mixture with a platinum-resistant chemoprotectant;

(b) first mixing a liquid solvent with a platinum-resistant chemoprotectant to produce an intermediate mixture, and then mixing the intermediate mixture with a gelling agent; or

(c) mixing a portion of the liquid solvent with a platinum-resistant chemoprotectant to produce a first mixture, mixing another portion of the liquid solvent with a gelling agent to produce a second mixture, and combining the first mixture and the second mixture .

The following examples are intended to illustrate the invention. They are not meant to limit the invention in any way.

Example

Example 1. Preparation of Thiosulfate Formulations

Hyaluronic acid gel 1 (0.5M STS, 1% (w/v) hyaluronic acid)

Sodium thiosulfate pentahydrate (619.75 mg) was dissolved in sterile distilled water (5 mL) in a sterile vial to yield a clear solution. Hyaluronic acid (50.30 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to the solution, and the resulting mixture was stirred at 4°C for 8-10 minutes. The resulting solution was filtered through a 0.22 μm Millex-GV sterile filter.

Hyaluronic Acid Gel 2 (0.1M STS, 2% (w/v) Hyaluronic Acid)

A15 Premium LV，Dow Chemical Company)溶于无菌蒸馏水(2.0mL)中，并将所得溶液与硫代硫酸钠溶液混合。将透明质酸(100.10mg；制药级80，Kikkoman Biochemifa company；0.6-1.2mDa)添加到所得混合物中，并在4℃下混合10-15分钟。Sodium thiosulfate pentahydrate (124.87 mg) was dissolved in sterile distilled water (3.031 mL). Methylcellulose (351.01 mg;

A15 Premium LV, The Dow Chemical Company) was dissolved in sterile distilled water (2.0 mL) and the resulting solution was mixed with the sodium thiosulfate solution. Hyaluronic acid (100.10 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to the resulting mixture and mixed at 4°C for 10-15 minutes.

Hyaluronic Acid Gel 3 (0.5M STS, 2% (w/v) Hyaluronic Acid)

A15 Premium LV，Dow Chemical Company)溶于无菌蒸馏水(2.0mL)中，并将所得溶液与硫代硫酸钠溶液混合。将透明质酸(100.65mg；制药级80，Kikkoman Biochemifa company；0.6-1.2mDa)添加到所得混合物中，并在4℃下混合10-15分钟。Sodium thiosulfate pentahydrate (620.35 mg) was dissolved in sterile distilled water (3 mL). Methylcellulose (350.23 mg;

A15 Premium LV, The Dow Chemical Company) was dissolved in sterile distilled water (2.0 mL) and the resulting solution was mixed with the sodium thiosulfate solution. Hyaluronic acid (100.65 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to the resulting mixture and mixed at 4°C for 10-15 minutes.

Hyaluronic Acid Gel 4 (0.1M STS, 1% (w/v) Hyaluronic Acid, Mannitol)

Hyaluronic acid (50.09 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to water (5 mL). Sodium thiosulfate pentahydrate (124.9 mg) was added. The pH of the resulting mixture was adjusted to pH 7.12 by the addition of sodium hydroxide (1 N, about 0.5 μL). An appropriate amount of mannitol was added to the vial to adjust the osmolality to 1.046 Osm/kg. The viscous solution was filtered through a 0.22 μm Millex-GV filter.

Hyaluronic acid gel 5 (0.1M STS, 1% (w/v) hyaluronic acid)

Hyaluronic acid gel 5 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide sodium thiosulfate at a concentration of 0.1M.

Hyaluronic Acid Gel 6 (0.2M STS, 1% (w/v) Hyaluronic Acid)

Hyaluronic acid gel 6 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide sodium thiosulfate at a concentration of 0.2M.

Hyaluronic acid gel 7 (0.3M STS, 1% (w/v) hyaluronic acid)

Hyaluronic acid gel 7 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide a 0.3M concentration of sodium thiosulfate.

Hyaluronic Acid Gel 8 (0.4M STS, 1% (w/v) Hyaluronic Acid)

Hyaluronic acid gel 8 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide a 0.4M concentration of sodium thiosulfate.

Hyaluronic Acid Gel 9 (0.5M STS, 1% (w/v) Hyaluronic Acid, Tris (5x))

Hyaluronic acid (79.99 mg; pharmaceutical grade 80, Kikkoman Biochemifa comp any; 0.6-1.2 mDa) was added to Tris buffer (8 mL, AMRESCO-0497-500G). The pH of the resulting mixture was adjusted to pH 7.13 by adding HCl (5N). Sodium thiosulfate pentahydrate (992.60 mg) was added to the above solution. The viscous solution was filtered through a 0.22 μm Millex-GV filter.

Hyaluronic Acid Gel 10 (0.5M STS, 1% (w/v) Hyaluronic Acid, Phosphate Buffered Saline (5x))

Hyaluronic acid (70.38 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to PBS buffer (7 mL, 5x). Sodium thiosulfate pentahydrate (868.46 mg) was added. The pH of the resulting mixture was adjusted to pH 6.99 by adding NaOH (IN). The viscous solution was filtered through a 0.22 μm Millex-GV filter.

Hyaluronic acid gel 11 (0.8M STS, 1% (w/v) hyaluronic acid)

Hyaluronic acid gel 11 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide a 0.8M concentration of sodium thiosulfate.

Hyaluronic Acid Gel 12 (1M STS, 0.8% (w/v) Hyaluronic Acid)

Hyaluronic acid gel 12 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide a 1 M concentration of sodium thiosulfate, and the amount of hyaluronic acid Adjusted to provide hyaluronic acid at a concentration of 0.8% (w/v).

Hyaluronic Acid Gel 13 (0.5M STS, 0.82% (w/v) Hyaluronic Acid (HYALGAN))

Hyaluronic acid gel 13 was prepared by mixing sodium thiosulfate pentahydrate with hyaluronic acid (HYALGAN, Fidia Pharma USA, Florham Park, NJ) to provide a hyaluronic acid concentration of 0.82% (w/v ) final preparation.

Hyaluronic Acid Gel 14 (0.5M STS, 1% (w/v) Hyaluronic Acid (SINGCLEAN))

Hyaluronic acid gel 14 was prepared according to the procedure described for hyaluronic acid gel 13, except that hyaluronic acid (SINGCLEAN, Hangzhou Singclean Medical Products Co., Ltd., Hangzhou, China) was used for the gel preparation.

Hyaluronic Acid Gel 15 (0.5M STS, 1% (w/v) Hyaluronic Acid (EUFLEXXA))

Hyaluronic acid gel 15 was prepared according to the procedure described for hyaluronic acid gel 13, except that hyaluronic acid (EUFLEXXA, Ferring Pharmaceuticals Inc., Parsippany, NJ) was used in the preparation of the gel .

Hyaluronic Acid Gel 16 (0.5M STS, 1% (w/v) Hyaluronic Acid (HEALON))

Hyaluronic acid gel 16 was prepared according to the procedure described for hyaluronic acid gel 13, except that hyaluronic acid (HEALON, Johnson & Johnson, New Brunswick, NJ) was used in the preparation of the gel.

Hyaluronic Acid Gel 17 (1M STS, 1% (w/v) Hyaluronic Acid)

Hyaluronic acid gel 17 was prepared according to the procedure described for hyaluronic acid gel 1, except that the amount of sodium thiosulfate pentahydrate was adjusted to provide a 1M concentration of sodium thiosulfate.

Hyaluronic Acid Gel 18 (10% (w/v) N-Acetyl-L-Cysteine, 1% (w/v) Hyaluronic Acid)

Hyaluronic acid (39.38 mg; pharmaceutical grade 80, Kikkoman Biochemifa company; 0.6-1.2 mDa) was added to water (4 mL). N-Acetyl-L-cysteine (399.14 mg) was added. The pH of the resulting mixture was adjusted to pH 7.21 by adding NaOH (10 N, 240 μL). The viscous solution was filtered through a 0.22 μm Millex-GV filter. The osmotic pressure was measured to be 1.107 Osm/kg.

Other hyaluronic acid gels can be prepared using the procedures described herein. For example, 1M and 1.5M hyaluronic acid gels can be prepared according to the same procedures as described for, eg, hyaluronic acid gel 1 and hyaluronic acid gel 12. Additionally, the pH level of the gel can be adjusted to pH 6.5 to 8.5 with brinast acid (eg, hydrochloric acid) and base (eg, sodium hydroxide).

Example 2. Pharmacokinetic Modeling of Perilymph Concentrations

Maximum cochlear platinum levels in humans following high dose cisplatin treatment (100 mg/m ² ) were modeled by using pharmacokinetic (PK) simulations of cisplatin distribution in humans. The kinetic parameters used for the simulations were obtained from population PK for cisplatin (Urien et al., Br. J. Clin. Pharmacol., 57:756-63, 2004) and PK after high-dose cisplatin treatment (Andersson et al., J.Pharm.Sci., 85:824-27, 1996) reported in the literature. PK simulations were performed using WinNonlin (Phoenix 64) PK simulation model 9 (intravenous infusion, 2 compartments). Based on the results of previous animal studies, a plasma to cochlear concentration ratio of 1:1 was assumed, and this assumption resulted from literature reports showing tissue distribution characteristics of cisplatin (Johnsson et al., Cancer Chemother. Pharmacol., 37:23- 31, 1995) for further support.

The predicted maximum plasma platinum concentration following cisplatin treatment was determined to be approximately 22 μM. Applying a 30-fold molar stoichiometric ratio resulted in a concentration of 660 μM (0.66 mM), which when the molecular weight of the thiosulfate was used, resulted in a thiosulfate concentration of 74 μg/mL. Achieving this thiosulfate level in the human cochlea is expected to provide complete (maximum) protection against cisplatin-induced ototoxicity following high dose (eg, 100 mg/m ² ) cisplatin treatment.

Example 3. In vivo pharmacokinetic studies

Hyaluronic acid gel No. 1 was administered to male Hartley guinea pigs at a dose of 12% w/v (0.5M, 6.2 mg). The dose volume was fixed (10 μL). Perilymph was sampled at each time point (n=5 animals/time point) and the concentration of thiosulfate was quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Hyaluronic acid gel 1 reached a maximum perilymph concentration of 868.5 μg/mL (about 7.8 mM) at the first sampling time point (1 h). This maximum perilymph concentration was approximately 10-fold higher than the concentration of Hyaluronic Acid Gel 1 (minimum effective concentration; 74 μg/mL, 0.66 mM) predicted to provide 100% protection from cisplatin-induced ototoxicity. Perilymph t _1/2 ranged from 2.7 hours to 6.4 hours. The combination of high perilymph hyaluronic acid gel 1 concentration and relatively long half-life provided a therapeutic window from 3 hours before cisplatin treatment to 4 hours after cisplatin treatment. In another pharmacokinetic study, healthy subjects were divided into 4 dose cohorts: Cohort 1, Cohort 2, Cohort 3 and Cohort 4. Each dose cohort comprised 8 human subjects randomized to receive DB-020 or placebo (6/2 randomized; 6 subjects received DB-020, 2 subjects received placebo). Group 1 was administered 19 mg of sodium thiosulfate in a 0.15M sodium thiosulfate/hyaluronic acid gel prepared as described for Hyaluronic Acid Gel 1, unilaterally. Group 2 was administered 62 mg of sodium thiosulfate as hyaluronic acid gel 1 unilaterally intratympanic. Group 3 was administered 124 mg of sodium thiosulfate as hyaluronic acid gel 17 unilaterally intratympanic. Group 4 was administered 186 mg of sodium thiosulfate, a 1.5 M sodium thiosulfate/hyaluronic acid gel prepared as described for Hyaluronic Acid Gel 1, unilaterally. Placebo subjects were administered 1% w/v of hyaluronic acid in water.

The results of the study are shown in Tables 1 and 2 and Figures 1-3.

Table 1

Table 2

^a increase to endogenous = (mean thiosulfate C _max ) - (mean thiosulfate placebo level)

^b MW of thiosulfate = 112 g/mol

Example 4. In vivo pharmacodynamic studies

To assess the therapeutic window of sodium thiosulfate gel in relation to cisplatin administration, 24 hours, 6 hours, 3 hours, or 1 hour before cisplatin administration, or 1 hour, 4 hours, or 24 hours after A single dose of 1.24 mg of hyaluronic acid gel was administered 1 intratympanic (IT) to the left ear of guinea pigs (single 10 mg/kg intravenous bolus; Figure 4). All control right ears not treated with Hyaluronic Acid Gel 1 exhibited a significant threshold shift (ie, hearing loss) compared to untreated animals (Figure 5A). Hyaluronic acid gel 1 administered from 3 hours before cisplatin administration to 4 hours after cisplatin administration provided protection against cisplatin-induced hearing loss relative to control ears not treated with hyaluronic acid gel 1 ( Figure 5B). Hyaluronic acid gel 1 was less effective in protecting against cisplatin-induced hearing loss when administered more distally to cisplatin treatment (eg, > 6 h before cisplatin administration or 24 h after cisplatin administration) Difference. The greatest protection was observed when hyaluronic acid gel 1 was administered 1 h before cisplatin administration, indicating that hyaluronic acid gel 1 can be achieved when hyaluronic acid gel 1 is administered before and preferably proximal to cisplatin treatment. The highest level of protection.

Example 5. Pharmacokinetic properties of exemplary hydrogels

Guinea Pig, Study 1

The study was performed using albino guinea pigs (Hartley) weighing 250-350 g. For dosing, the animal was placed against its shoulder with the surgical ear facing up and the auditory bulb was first exposed using a retroauricular approach. A hole with a diameter of 2-3mm was drilled in the ear bulb to provide direct observation of the microenvironment of the round window. Then, 10 μL of the aqueous composition of 0.5M sodium thiosulfate/2% (w/v) hyaluronic acid (STS composition) was applied to the RWM using a 10 μL Hamilton syringe and 26 gauge needle. Following application, the guinea pigs were kept in this position for 30 minutes to allow the compound to diffuse into the cochlea. The auditory bulb opening was closed with a muscle graft, and the incision was closed with sutures.

Briefly, the sampling procedure is as follows. All sampling procedures are end-point. Euthanize animals with _CO . A 0.5 mL blood sample was collected by cardiac puncture. Plasma was separated by centrifugation at 5,000 rpm for 10 minutes at 4°C and collected in separate tubes. Collect 50 μL of cerebrospinal fluid through the cisterna magna. Perilymph was collected ex vivo to avoid contamination from cerebrospinal fluid inflow via the cochlear aqueduct. The temporal bone was rapidly isolated, and the auditory bulb was removed to expose the cochlea. Any visible residual dose composition was carefully removed with an absorbent point under a surgical microscope prior to perilymph sampling. A small hole was punched at the tip and 5-7 μL of perilymph was sampled using a pulled glass pipette. All samples were immediately frozen on dry ice and stored at -80°C until analysis. The concentration of thiosulfate in the sample was measured using the method disclosed in Togawa et al., Chem. Pharm. Bull., 40:3000-3004, 1992, the disclosure of which is incorporated herein by reference. The results of this study are shown in Figures 8A, 8B, 9A, 9B and Table 3.

cynomolgus monkey

Tovudine (4 mg/kg) was administered subcutaneously to cynomolgus monkeys. After 30 minutes, animals were anesthetized by an intravenous bolus of propofol (5.5 mg/kg). Animals were then maintained under anesthesia using 2-3% isoflurane inhalation. The animal was then immobilized and placed laterally in the reverse Trendelenburg position to ensure access to the round window. Animals were kept on a warm blanket during the surgical procedure.

Intratympanic injections were performed in the right ear when the animals reached anesthesia. 1.1 mL of epinephrine hydrochloride-saline (0.1 mg in 10 mL of saline) and 0.5 mL of lidocaine hydrochloride (20 mg/mL) were injected subcutaneously into the skin of the back wall of the ear canal in each ear as local anesthetics, respectively. . An incision is then made in the skin behind the pinna and a portion of the temporal bone is drilled to expose the middle ear. 50 μL of the STS composition was injected into the round window membrane using a 25G needle. Following dosing, the animals were placed in a straight line, head up, to allow the dosing solution to remain in the tympanic chamber for 30 minutes. Then repeat the same process for the other ear.

Plasma and CSF were collected approximately 2 hours after dosing in the first ear (right). Right cochlear pericymph sampling was performed approximately 3 hours after dosing in the right ear. Animals were euthanized by intravenous injection of 11 mg/kg of propofol and then bled via the femoral artery. The animals were then placed in a lateral recumbent position. A post-auricular skin incision was performed, and the external auditory canal was removed to expose the middle ear. A portion of the temporal bone is then drilled to expose the basal turn of the cochlea. Clean the remaining dose (if visible) in the middle ear with a cotton swab. A drop of tissue glue was applied at the base of the cochlea to minimize contamination from the administered composition.

Using a 0.5-1mm burr crochet or sharp crochet, make a hole in the bottom turn of the cochlea. Perilymph (approximately 10 μL) was then collected using a capillary tube inserted into the scala tympanum of the cochlea. The same procedure was repeated for left cochlear pericymph sampling approximately 2 hours after administration to the left ear. The results of this study are shown in Table 3.

table 3

In the above table, IT is intratympanic administration and TT is transtympanic administration.

* Thiosulfate concentration measured from plasma samples of test animals.

Guinea Pig, Study 2

Male guinea pigs of about 5-7 weeks old weighing 200-300 g were used as subjects (N=5 per group). Prior to any procedure, animals were anesthetized with Zolazepam hydrochloride (sutai 50; 20 mg/kg) via the intramuscular route 10 minutes before surgery. If needed, an intraoperative booster at one-tenth of the original dose will be administered intraperitoneally.

Intratympanic injection:

1. Under microscope magnification, use sharp scissors to make a 0.5-1.5 cm post-auricular skin incision at approximately 6-8 mm caudal to the ear-head crease. Be careful to avoid cutting too deep to protect the underlying vascular structures.

2. Perform blunt dissection with forceps carefully through the subcutaneous fat layer, muscle and tissue. Gently retract the mastoid body until the smooth dome of the periosteum is seen. At the caudal portion of the auditory bulb, the deeper cervical muscle, where the pectoralis muscle attaches, can be seen. The facial nerve, which became visible at the dorsal and snout of the auditory bulb dome, was preserved during surgery.

3. A self-retaining retractor was placed in the posterior part of the auditory bulb, before a small hole (0.5 mm in diameter) was made using a drill. Use a pair of jewel-tipped forceps to cut the ossicles in the dorsal and caudal direction. Under high magnification, the bones are removed one by one. Care was taken not to puncture the stapedial artery, which lies directly under the bulla cap, as bleeding from this artery could jeopardize the surgical procedure. The amount of bone removed was kept to a minimum to prevent excess fluid from entering the middle ear, while still allowing for good visualization and proximity to the round window microenvironment.

4. Use a sterile glass Hamilton syringe with a 25-26G blunt needle to deliver 10 or 90 μL of the gel formulation into the round window microenvironment.

5. Allow the delivered agent to reside within the round window microenvironment for up to 30 minutes. This small hole is covered with muscle tissue and tissue glue.

6. The incision is closed with sutures (4-0 nonabsorbable monofilament or 5-0 nonabsorbable nylon) and tissue glue or wound clips. The entire procedure takes about 3-5 minutes, depending on the dosage specifications.

7. During surgery and until recovery, animals are placed on a temperature-controlled (38°C) heating pad until consciousness is regained, at which point the animals are returned to their home-cage.

Alternatively, the gel formulation is administered to the animal via the tympanic cavity.

Sample collection:

Blood collection:

1. Put the guinea pigs in the euthanasia box without pre-inflation in the euthanasia box and introduce 100% carbon dioxide to make the animal unconscious and reduce the suffering of the animal. The carbon dioxide flow was maintained for a minimum of 1 minute after breathing ceased. After confirmation of death, the guinea pigs were removed from the euthanasia box.

2. Blood was collected immediately after euthanasia.

3. After the operator has secured the animal in the dorsal position, the needle is inserted 4-6 or slightly forward at the front of the sternal ridge.

4. Pull the needle back and the blood returns.

5. Volume: For each blood collection, approximately 1 mL of blood is collected.

CSF Collection:

CSF was collected after euthanasia. Slowly lower the 0.5*20 IV needle at 90° relative to the foramen magnum. Allow the needle to reach a distance of 4.5-5 mm under the skin and remove 50-200 μl of clear tissue fluid.

Perilymph collection:

After euthanasia, animals were stripped of excess skin and muscle tissue to obtain intact auditory bulbs, and the walls of the auditory bulbs were cut open with small forceps to expose the cochlea. Use a small cotton ball to clean the bottom of the foam. Cochlear bottom ring and round window were coated with bioglue. After drying, a single micro-hole was manually drilled at the apical ring of the cochlea. 2 μL of perilymph was then collected with a microcapillary inserted into the cochlear top ring. Perilymph samples were added to vials containing 18 [mu]L bovine serum albumin (BSA, 1M) and stored at -80&lt;0&gt;C until analysis.

The results of Guinea Pig Study 2 are provided in Tables 4 and 5.

Table 4

In this table, TT is administered via the tympanic membrane,

*This test is a repetition of the previous test.

table 5

In this table, IT is intratympanic administration and TT is transtympanic administration.

Example 6. Pharmacodynamic properties of exemplary hydrogels

Cisplatin was diluted with 0.9% (w/v) saline to a final concentration of 5 mg/mL. Albino guinea pigs (Hartley) weighing 250-350 g were used for the study. After at least 3 days of acclimation, 28 animals were included in the study. Cisplatin is administered intraperitoneally using a bolus injection under sterile conditions. The five cohorts were staggered with different start dates for the study.

Seven days after cisplatin administration, the animals' auditory brainstem responses (ABR) were recorded using a TDT RZ6 multiple input multiple output processor. Baseline was defined using historical ABR data. Animals were anesthetized with teletamine hydrochloride and zolazepam hydrochloride (sutai). Acoustic stimuli are delivered via earphones. Needle electrodes were placed at the caudal-ventral location near the ear canal, the apex of the skull, and the ground of the lower leg. Stimulus levels were from 10 dB to 90 dB in 5 dB steps, and the short-tone frequencies were 4 kHz, 24 kHz and 32 kHz. The upper limit of the sound pressure level is 90dB. The ABR threshold was observed by visually inspecting the stacked waveforms where the waveforms were above the noise floor as the lowest sound pressure level.

Before the cisplatin study, ABR data were recorded for each animal's ears from 50 animals (n=100 without experimental treatment). Untreated animals had a threshold of 39.8dB at 32kHz. The normal hearing range was defined as mean ±2SD, 27.9 to 51.6dB. Cisplatin induces hearing loss primarily at high frequencies. Clear patterns of hearing loss following cisplatin were defined as thresholds of 60 dB and higher at 32 kHz.

In this study, 1 of 28 animals died before measurement day 7. Of the remaining 27 animals, 18 animals had hearing loss with a use threshold >60dB at 32kHz (Figure 10A). Hearing loss at 32kHz ranged from a threshold of 65dB to 90dB (Figure 10B). 90dB is the upper limit of measurement. It should be noted that the threshold is limited to 90dB when there is no waveform or only a waveform is seen at 90dB. The average threshold at 32kHz is 82dB, which corresponds to an average 42.2dB shift from the initial threshold of 39.8dB (Fig. 11A).

Local intratympanic drug delivery and cochlear sampling

Local intratympanic administration and cochlear sampling were performed as described in Example 5.

Topically delivered anti-platinum chemoprotectants provide hearing protection from platinum-based antineoplastic agents.

Evaluation of the hearing protective effect of locally delivered anti-platinum chemoprotectants against platinum-based antineoplastic agents was performed as follows.

An aqueous composition of 0.5M sodium thiosulfate/2% (w/v) hyaluronic acid (STS composition) or vehicle was intratympanically applied to the round window in the left ear (LE) as described above, and The right ear (RE) of the guinea pig was left untreated (Figure 12). Sixty minutes after STS composition or vehicle administration, animals were injected intraperitoneally with 10 mg/kg of cisplatin. Seven days after cisplatin administration, ABR was measured at 4 kHz, 24 kHz and 32 kHz in both ears.

Due to the heterogeneity of hearing loss following cisplatin challenge, the untreated right ear was used to select animals with hearing loss. There were 21 animals with a right ear threshold >60dB at 32kHz. Of these 21 animals, 3 animals with otitis media were excluded, leaving 18 animals for final analysis. 10 animals were dosed with vehicle, and 8 animals were dosed with the STS composition (FIG. 11B). In the untreated right ear, there was no difference in ABR thresholds between the STS composition and vehicle groups, with a mean threshold of 73dB at 4kHz, 71dB at 24kHz, and a mean threshold at 32kHz is 80dB. The vehicle-treated left ear was not significantly different from the untreated right ear, showing a threshold of 74 dB at 4 kHz, 70 dB at 24 kHz, and 74 dB at 32 kHz.

STS composition-treated ears had significantly lower thresholds at both 32KHz and 24KHz compared to vehicle-treated ears and untreated right ears (***P<0.001, two-way ANOVA). At 4 kHz, the mean threshold was 61 dB in the STS composition-treated ear and 75 dB in the untreated contralateral right ear; protection was not statistically significant (P=0.089). Mean thresholds at 24 kHz and 32 kHz in ears treated with the STS composition were 40 dB and 48 dB, respectively, compared to 69 dB and 80 dB, respectively, in the contralateral untreated right ear. Normal hearing thresholds in untreated animals were 35dB and 40dB at 24kHz and 32kHz, respectively. For ears of non-experimentally treated animals, the untreated ears after cisplatin had an average of 34 dB and 40 dB of threshold rise at 24 kHz and 32 kHz, respectively, but the STS composition-treated ears had only 5 dB and 8 dB of excursion. Therefore, sodium thiosulfate provides an average of 80% protection at both 24 kHz and 32 kHz.

In a similarly designed study as described above, sound pressure levels at 4 kHz, 24 kHz and 32 kHz were measured during ABR tests on guinea pigs administered vehicle or sodium thiosulfate (0.1 M per ear) , 0.5M or 1M sodium thiosulfate gel) followed by a cisplatin challenge (10MPK of cisplatin, i.v.). Different doses of hyaluronic acid gel were administered as 10 μL IT injections into the left ear 1 hour before cisplatin administration. The animal's contralateral ear (right ear) was left untreated. Hyaluronic Acid Gel 5 (0.1 M), Hyaluronic Acid Gel 1 (0.5 M) and Hyaluronic Acid Gel 17 (1 M) were tested. Untreated ears showed a clear threshold shift compared to untreated animals (grey shaded area). At all frequencies tested, the groups treated with Hyaluronic Acid Gel 1 (0.5 M) and Hyaluronic Acid Gel 17 (1 M) showed hearing protection compared to the untreated contralateral control ear. No protection was seen in vehicle-treated ears. The results are summarized in Figure 13.

Other embodiments

Various modifications and variations of the described invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention will be apparent to those skilled in the art and are intended to be included within the scope of the invention.

Other embodiments are within the scope of the following claims.

Claims (20)

1. A method of reducing platinum-induced ototoxicity in a subject in need, the method comprising administering to the subject an effective amount of thiosulfate, wherein the subject is administered the The platinum-based neoplastic agent was administered no more than 7 hours prior to the thiosulfate, or the platinum-based antineoplastic agent was scheduled to be administered within 4 hours. 2. The method of claim 1, wherein the subject is administered a platinum-based tumor agent no more than 7 hours prior to administration of the thiosulfate. 3. The method of claim 1, wherein the subject is scheduled to be administered a platinum-based antineoplastic agent within 4.5 hours. 4. The method of claim 1, wherein the subject is administered a platinum-based tumor agent no more than 2.5 hours prior to administration of the thiosulfate. 5. The method of claim 1, wherein the subject is administered a platinum-based tumor agent no more than 1 hour prior to administration of the thiosulfate. 6. The method of claim 1, wherein the thiosulfate is administered otically. 7. The method of claim 6, wherein the thiosulfate is administered intratympanically, transtympanically, or by inner ear injection. 8. The method of claim 1, further comprising administering the platinum-based antineoplastic agent. 9. The method of claim 1, wherein the thiosulfate salt is a basic thiosulfate salt, an ammonium thiosulfate salt, or a solvate thereof. 10. The method of claim 1, wherein the effective amount of thiosulfate is administered as a hypertonic pharmaceutical composition comprising the effective amount of thiosulfate. 11. The method of claim 10, wherein 200-1,000 [mu]L of the hypertonic pharmaceutical composition is administered to the subject's round window. 12. The method of claim 10, wherein the calculated osmolality of the hypertonic pharmaceutical composition is 500-5,000 mOsm/L. 13. The method of claim 10, wherein the concentration of the thiosulfate in the hypertonic pharmaceutical composition is 0.5M-2.5M. 14. The method of claim 1, wherein the effective amount is an amount that produces a plasma thiosulfate concentration of 30 [mu]M or less upon administration of the platinum-based antineoplastic agent. 15. The method of claim 1, wherein the effective amount is 0.1-2.5 mmol of the thiosulfate. 16. The method of claim 1, wherein the effective amount is an amount that produces a maximum thiosulfate concentration of 0.6-10 mmol/L 1 h after administration. 17. The method of claim 1, wherein the effective amount is an amount that produces a thiosulfate concentration of 0.1-2 mmol/L 7 h after administration in the cochlea of the subject. 18. A method of alleviating platinum-induced ototoxicity in a subject in need thereof, the method comprising administering to the subject an effective amount of thiosulfate to produce (i) a A plasma thiosulfate _Cmax of 30 μM or less for a neoplastic agent, and (ii) a cochlear thiosulfate _Cmax that is at least 30 times greater than the cochlear _Cmax of the platinum-based antineoplastic agent, wherein all The cochlear platinum concentrations and the cochlear _Cmax were modeled by pharmacokinetic simulations of intravenous infusion in a two-compartment model. 19. The method of claim 18, wherein the effective amount of thiosulfate is administered within 3 hours prior to administration of the platinum-based antineoplastic agent or within 4 hours after administration of the platinum-based antineoplastic agent. 20. The method of claim 18, wherein the effective amount of thiosulfate is administered within 4.5 hours before administration of the platinum-based antineoplastic agent or within 2.5 hours after administration of the platinum-based antineoplastic agent.

Images