System and method for producing hydroxypropyl-beta-cyclodextrin

Abstract

This specification provides a system and method for manufacturing hydroxypropyl-β-cyclodextrin.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the priority of U.S. Provisional Application No. 63 / 351,719, filed on June 13, 2022, entitled "SYSTEMS AND METHODS FOR MANUFACTURING HYDROXYPROPYL - BETA - CYCLODEXTRIN", and incorporates the entire content thereof herein by reference.

[0002] This disclosure relates to systems and methods for manufacturing hydroxypropyl - beta - cyclodextrin. Accordingly, the disclosure broadly relates to the fields of chemistry, pharmacy, and chemical engineering.

Background Art

[0003] Hydroxypropyl - beta - cyclodextrin (HPBCD) has been gaining interest in the pharmaceutical field because it has the potential to treat multiple types of diseases. Therefore, new systems and methods for producing HPBCD are needed to meet the increasing demand.

Summary of the Invention

[0004] A reactor system for producing hydroxypropyl - beta - cyclodextrin (HPBCD) is provided herein. The system includes a propylene oxide feed, a beta - cyclodextrin (BCD) feed, a mass flow meter or mass flow controller, and a static mixer. In some embodiments, the system further includes a back - pressure regulator. In some additional embodiments, the system includes a mass flow controller. In still further embodiments, the system includes a temperature controller. In some aspects, the static mixer is a helical static mixer.

[0005] In some embodiments, the propylene oxide feed is pressurized. In other embodiments, the BCD feed is pressurized.

[0006] In some embodiments, the system comprises at least two propylene oxide feeds. In some aspects, at least two propylene oxide feeds are operably connected to separate mass flow meters or controllers. In some embodiments, the first propylene oxide feed supplies BCD at a concentration of about 7 to about 15 equivalents, and the second propylene oxide feed supplies BCD at a concentration of about 3.5 to about 15 equivalents.

[0007] In some embodiments, the BCD feed contains sodium hydroxide (NaOH). In some aspects, the β-cyclodextrin feed contains NaOH at a concentration of about 5 to about 10 equivalents.

[0008] In some embodiments, the system further comprises a pump. In some aspects, the pump can be a syringe pump operably connected to one or more of the feeds.

[0009] In some embodiments, the system further comprises a coiled tube. In some embodiments, the system comprises a plug flow reactor. In some aspects, the plug flow reactor includes at least two coiled tubes and a temperature control unit. In some embodiments, a back pressure regulator is operably connected to the plug flow reactor or the coiled tube. In some aspects, the temperature control unit maintains a temperature of about 30°C to about 60°C.

[0010] In some embodiments, propylene oxide is provided at two locations. In some aspects, propylene oxide is provided before the plug flow reactor. In some additional aspects, at least one feed amount of propylene oxide is provided before the first coiled tube, and at least another feed amount of propylene oxide is provided before the second coiled tube.

[0011] In some embodiments, the system further comprises a collection tank. In some aspects, the collection tank is operably connected to the acid feed. In some further aspects, the acid feed comprises hydrochloric acid, sulfuric acid, lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, malic acid, fumaric acid, tartaric acid, or combinations thereof. In some additional aspects, the system provides a total residence time of from about 30 minutes to about 70 minutes.

[0012] Further provided herein is a method for producing a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, comprising: (a) contacting a hydroxypropyl-β-cyclodextrin (HPBCD) mixture with at least two solvents, wherein the HPBCD mixture comprises highly substituted HPBCD and lowly substituted HPBCD; (b) dissolving the highly substituted HPBCD in one of the solvents; and (c) removing the lowly substituted HPBCD by precipitation. In some embodiments, the at least two solvents comprise ethanol and acetone.

[0013] Further provided herein is a method for producing a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, comprising: (a) contacting a hydroxypropyl-β-cyclodextrin (HPBCD) mixture with at least two solvents, wherein the HPBCD mixture comprises highly substituted HPBCD; (b) dissolving the highly substituted HPBCD in one of the solvents to form a mother liquor; and (c) filtering off the mother liquor. In some embodiments, the method comprises lyophilizing the mother liquor to obtain a solid. In some aspects, the method further comprises analyzing the solid by MALDI-TOF to determine the degree of substitution. In some embodiments, the at least two solvents comprise ethanol and acetone.

[0014] Further provided herein is a composition comprising a methylated 2-hydroxypropyl-β-cyclodextrin (HPBCD) mixture having a degree of substitution of about 6.5 to about 9.5 and a methylated glucose having 0 to 5 2-hydroxypropyl groups. In some examples, the composition may have a mass spectrum as shown in FIG. 25.

[0015] Further provided herein is a method for oligomer substitution by methanolysis of a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, the method comprising: (a) mixing HPBCD and methanol; (b) stirring until the HPBCD is dissolved; (c) adding an acid to the mixture; (d) heating the mixture to at least 50 to about 90 °C; (e) stirring the mixture and maintaining the heat for at least about 24 hours; (f) neutralizing the mixture with a base; and (g) filtering the mixture.

[0016] Further provided herein is a method for purifying a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, the method comprising: (a) purifying the HPBCD mixture by nanofiltration; (b) collecting a total of at least 5 volumes of nanofiltration permeate for diafiltration; and (c) lyophilizing the resulting residue to obtain solid hydroxypropyl-β-cyclodextrin.

[0017] In some embodiments, the purification is performed at a feed pressure of about 200 to about 400 psi (e.g., about 300 psi). In some embodiments, the purification by nanofiltration involves a flat sheet membrane. In some aspects, the flat sheet membrane has an area of 0.010 to 0.050 m 2 including.

[0018] In some embodiments, the method involves collecting a total of at least 7 volumes of nanofiltration permeate for diafiltration, or more preferably a total of at least 10 volumes of nanofiltration permeate for diafiltration.

[0019] Further provided herein is a method for purifying a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, the method comprising: (a) purifying the HPBCD mixture by nanofiltration; (b) collecting at least a total of five diafiltration volumes of the nanofiltration permeate; and (c) analyzing the propylene glycol content of the resulting residue. In some embodiments, the method further comprises lyophilizing the resulting residue to obtain solid hydroxypropyl-β-cyclodextrin.

[0020] In some embodiments, the claimed invention also encompasses a composition, including a composition produced according to any of the methods or systems described herein. For example, the composition may include a mixture of beta-cyclodextrin molecules substituted at one or more hydroxyl positions by hydroxypropyl groups, the mixture including less than 0.3% unsubstituted beta-cyclodextrin ("DS-0") or less than 1% beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The present invention includes a composition produced by the method according to any one of Embodiments 26 to 30 or 33 to 42, or any one of Claims 28 to 32 or 35 to 45, the composition including a mixture of beta-cyclodextrin molecules substituted at one or more hydroxyl positions by hydroxypropyl groups, the mixture including less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and at least 70% of the beta-cyclodextrin is DS aCompositions may also be included that have a DS within ±1σ, where σ is the standard deviation. Further, the present invention includes a composition produced by the method according to any one of Embodiments 26 - 30 or 33 - 42 or any one of Claims 28 - 32 or 35 - 45 and containing a mixture of beta - cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture contains less than 1% unsubstituted beta - cyclodextrin ("DS - 0") and beta - cyclodextrin substituted with 1 hydroxypropyl group ("DS - 1"), and the mixture contains 1% - 10% beta - cyclodextrin substituted with 7 hydroxypropyl groups ("DS - 7").

[0021] Alternatively, the composition is produced by any of the methods described herein (such as the methods described in any one of Embodiments 28-32 or 35-45), and comprises a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 25% or less of beta-cyclodextrin substituted with four hydroxypropyl groups ("DS-4"). Similarly, the present invention includes a composition produced by any one of Embodiments 26-30 or 33-42 or any one of Claims 28-32 or 35-45, and comprises a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 20% or less of beta-cyclodextrin substituted with five hydroxypropyl groups ("DS-5"). In another aspect, the composition is produced by any one of Embodiments 26-30 or 33-42 or any one of Claims 28-32 or 35-45, and comprises a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 2.5% of beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof.The composition is produced by the method according to any one of Embodiments 26 to 30 or 33 to 42 or any one of Claims 28 to 32 or 35 to 45, and includes a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture includes less than 1% of unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture includes 5% to 25% of beta-cyclodextrin substituted with six hydroxypropyl groups ("DS-6"). Optionally, the composition may be as described above. Finally, the present invention also provides a composition produced by the method according to any one of Embodiments 26 to 30 or 33 to 42 or any one of Claims 28 to 32 or 35 to 45, and includes a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture includes less than 1% of unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and also includes the following structure:. [Chemical formula] The composition may also include beta-cyclodextrin having glucose units of the formula, wherein R1, R2, and R3 are each independently -H or -HP at each occurrence, HP includes one or more hydroxypropyl groups, and the percentage of the combination of R1 and R2 that are HP in the beta-cyclodextrin ranges from 85% to 95% of the total occurrences. In some embodiments, the present invention simultaneously produces at least two different, at least three different, at least four different, at least five different compositions, where each composition includes a mixture of different beta-cyclodextrin molecules. Thus, in some embodiments, the present invention simultaneously produces at least a plurality of different compositions, where each composition includes a mixture of different beta-cyclodextrin molecules. BRIEF DESCRIPTION OF THE DRAWINGS

[0022]

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DETAILED DESCRIPTION OF THE INVENTION

[0023] This specification provides a reactor system for producing hydroxypropyl-β-cyclodextrin (HPBCD). Referring to FIG. 1, the reactor system 100 of the present disclosure generally includes a propylene oxide feed 102, a β-cyclodextrin (BCD) feed 104, a mass flow meter 106 or a mass flow controller 108, and a static mixer 110. The propylene oxide from the propylene oxide feed 102 and the BCD from the BCD feed 104 are combined and mixed in the static mixer 110. Next, the reactants pass through the reactor 118 to form a first reactor effluent, and then, if appropriate, additional propylene oxide is added from the second propylene oxide feed 102. This mixture passes through the second static mixer 110 and then enters the second reactor 118 to form a second reactor effluent. The second reactor effluent 118 is collected in the collection tank 124, where it is quenched with an acid supplied by the acid feed 126. The reactor system described herein is operable to efficiently produce HPBCD with a target degree of substitution.

[0024] The reactor system of the present disclosure is operable to produce HPBCD according to the reaction scheme shown below. BCD reacts with propylene oxide and a base (e.g., sodium hydroxide) and is then quenched with an acid (e.g., hydrochloric acid).

Chemical formula

[0025] System 100 includes at least one propylene oxide feed 102. However, it should be noted that the reactor system may include at least two propylene oxide feeds (i.e., at least a plurality of propylene oxide feeds), at least three propylene oxide feeds, and the like. The propylene oxide feed 102 may include a tank having conduits and instruments operable to deliver propylene oxide to the system 100. Propylene oxide may be introduced into the system at one or more locations. Propylene oxide may be introduced at a flow rate of from about 0.1 g / min to about 10 g / min, for example, about 0.1 g / min, 0.2 g / min, 0.3 g / min, 0.4 g / min, 0.5 g / min, 0.6 g / min, 0.7 g / min, 0.8 g / min, 0.9 g / min, 1.0 g / min, 2.0 g / min, 3.0 g / min, 4.0 g / min, 5.0 g / min, 6.0 g / min, 7.0 g / min, 8.0 g / min, 9.0 g / min, or about 10.0 g / min. Propylene oxide may be provided at one or more locations within the system 100. In a system 100 having more than one reactor 118, propylene oxide may be provided in front of each reactor. For example, as in the system 100 of FIG. 1, propylene oxide may be provided at two locations. However, generally, at least one supply amount of propylene oxide is provided in front of the reactor 118. In some embodiments, the propylene oxide feed may include a racemic mixture of propylene oxide, and in other embodiments, the propylene oxide feed may include enantiopure propylene oxide. Propylene oxide may include deuterated propylene oxide.

[0026] Propylene oxide can be provided at a concentration of about 1 to about 20, about 3.5 to about 20, about 5 to about 20, about 7 to about 20, about 1 to about 15, about 3.5 to about 15, about 5 to about 15, or about 7 to about 15 molar equivalents of BCD. For example, propylene oxide can be provided at a concentration of about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, or about 20 molar equivalents of BCD. In embodiments where propylene oxide is provided at two locations, the first propylene oxide feed may supply propylene oxide at a concentration of about 7 to about 15 molar equivalents of BCD, and the second propylene oxide feed may supply propylene oxide at a concentration of about 3.5 to about 15 molar equivalents of BCD.

[0027] System 100 includes at least one BCD feed 104. The BCD feed 104 can include a tank having conduits and apparatus operable to deliver BCD to the system 100. BCD can be introduced at a flow rate of about 0.0 g / min to about 20 g / min, about 0.1 g / min to about 10 g / min, about 0.5 g / min to about 7 g / min, or about 1.0 g / min to about 5 g / min, for example, about 0.1 g / min, about 0.2 g / min, about 0.3 g / min, about 0.4 g / min, about 0.5 g / min, about 0.6 g / min, about 0.7 g / min, about 0.8 g / min, about 0.9 g / min, about 1.0 g / min, about 2.0 g / min, about 3.0 g / min, about 4.0 g / min, about 5.0 g / min, about 6.0 g / min, about 7.0 g / min, about 8.0 g / min, about 9.0 g / min, or about 10.0 g / min. The BCD feed can include deuterated BCD.

[0028] The system can further include a base or sodium hydroxide (NaOH) feed. The base or sodium hydroxide can be supplied at a concentration of about 1 to about 10, about 3 to about 10, about 5 to about 10, or about 7 to about 10 molar equivalents of BCD, or more preferably at a concentration of about 5 to about 10 molar equivalents of BCD. In some embodiments, the BCD feed can include the base or sodium hydroxide.

[0029] The propylene oxide feed(s) 102 and / or the BCD feed(s) 104 may be pressurized. Pressurizing the feed can be beneficial when low flow rates (e.g., about 1.5 g / min) of reactants are required. The feed may be pressurized with an inert gas such as a noble gas (e.g., helium, neon, argon, krypton, or xenon), or another non-reactive gas such as nitrogen or carbon dioxide. The inert gas may be supplied within a pressurized tank 114 operably connected to the feed.

[0030] The propylene oxide feed(s) 102 and / or the BCD feed(s) 104 may be operably connected to a mass flow meter 106. The mass flow meter 106 is operable to determine the mass flow rate of propylene oxide or BCD. Mass flow meters and methods of measuring mass flow rate are well known in the art. Additional mass flow meters may be included at other locations within the system to monitor the mass flow rate of reactants and / or products.

[0031] The propylene oxide feed(s) 102 and / or the BCD feed(s) 104 may be operably connected to a mass flow controller 108. The mass flow controller is operable to control the mass flow rate of propylene oxide or BCD, e.g., the mass flow controller can increase, decrease, or hold constant the mass flow rate of the feed. Mass flow controllers and methods of measuring mass flow rate are well known in the art.

[0032] The mass flow meter(s) 106 and / or the mass flow controller(s) 108 may be operably connected to the controller. The controller may be operable to communicate electronically or wirelessly with any of the system components. Generally, the controller may include one or more processors and a non-transitory computer-readable storage medium storing instructions for causing one or more of the processors to control one or more of the start-up, operation, or stop of any one or more of various aspects of the system to facilitate safe and efficient operation. For example, the controller may cut off power to any of the system components if an abnormal condition is detected. The controller may also be operable to open or close valves or to adjust other system parameters (such as temperature and pressure) to ensure safe and efficient operation of the system.

[0033] System 100 may further include at least one static mixer 110. The static mixer is operable to continuously mix the fluid flowing through the static mixer without using moving parts by inducing the flow to increase turbulence. Static mixers are well known in the art and may include plates, baffles, helical elements, or geometric grids. In an exemplary embodiment, the static mixer is a helical static mixer. System 100 may include one or more static mixers 110 at various locations within system 100.

[0034] One or more of the feeds may be operably connected to a pump 116. The pump may be any pump known in the art, including centrifugal pumps, positive displacement pumps, syringe pumps, etc. Pump 116 may be operably connected to one or more of the feeds. In an exemplary embodiment, the system includes a syringe pump operably connected to a BCD feed.

[0035] System 100 may further include a reactor 118. The reactor includes a plug flow reactor that may include at least one coiled tube. In some embodiments, the system may include two or more reactors. In additional embodiments, the plug flow reactor may include at least two coiled tubes. The reactor may have a volume of about 1 to about 1000 mL, about 1 to about 500 mL, about 1 to about 250 mL, or about 1 to about 100 mL, such as about 1 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, about 10 mL, about 20 mL, about 30 mL, about 40 mL, about 50 mL, about 60 mL, about 70 mL, about 80 mL, about 90 mL, about 100 mL, about 250 mL, about 500 mL, or about 1000 mL. The reactor may also have a volume greater than 100 mL, greater than 250 mL, greater than 500 mL, or greater than 1000 mL.

[0036] Using the volume of the reactor and the flow rate of the reactants, the residence time of the reactants in the reactor can be determined. The reactants may have a residence time in the reactor of about 1 minute to about 360 minutes, about 3 minutes to about 180 minutes, about 5 minutes to about 90 minutes, or about 10 minutes to about 60 minutes, such as about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about 60 minutes. The reactants may have a residence time in the reactor greater than 60 minutes, greater than 90 minutes, greater than 180 minutes, or greater than 360 minutes.

[0037] System 100 may further include a temperature control unit 122. The temperature control unit may be operably connected to one or more reactors 118. The temperature control unit 122 may maintain a temperature in the reactor(s) 118 of about 40°C to about 50°C, about 35°C to about 55°C, about 30°C to about 60°C, about 25°C to about 65°C, about 20°C to about 70°C, about 15°C to about 90°C, or about 10°C to about 95°C.

[0038] System 100 may further include a backpressure regulator 112. The backpressure regulator is operable to maintain a predetermined set pressure upstream of the backpressure regulator. Generally, the backpressure regulator 112 is disposed near an end of the system 100, for example, immediately before the collection tank 124. Accordingly, the backpressure regulator may be operably connected to the collection tank 124. The backpressure regulator may also be operably connected to the reactor. The backpressure regulator may be operable to maintain a backpressure of about 0 psi to about 500 psi, about 1 psi to about 400 psi, about 1 psi to about 300 psi, about 3 psi to about 200 psi, about 5 psi to about 100 psi, about 10 psi to about 50 psi, for example, about 10 psi, about 15 psi, about 20 psi, about 25 psi, about 30 psi, about 35 psi, about 40 psi, about 45 psi, or about 50 psi. The backpressure regulator may be operable to maintain a backpressure greater than 5 psi, greater than 10 psi, greater than 25 psi, greater than 50 psi, greater than 100 psi, greater than 200 psi, greater than 300 psi, greater than 400 psi, or greater than 500 psi.

[0039] System 100 may further include a collection tank 124. The collection tank may be operable to hold products and / or residual reactants from the reaction. Further, the collection tank may be operable to quench the mixture from the reactor 118 with an acid such as hydrochloric acid. The acid may be fed stoichiometrically with respect to the HPBCD produced or in an amount sufficient to reach a predetermined pH. The contents of the collection tank generally include a crude HPBCD mixture. Collection tanks are well known in the art. The collection tank may further include a stirring mechanism for continuously stirring the contents to maintain a homogeneous mixture.

[0040] System 100 may further comprise an acid feed 126. The acid feed 126 may be operably connected to the collection tank 124. The acid feed may include hydrochloric acid, sulfuric acid, lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, malic acid, fumaric acid, tartaric acid, or combinations thereof. Alternatively, the reactor effluent may be quenched by contacting it with an acidic ion exchange resin such as Amberlyst™ 35 Dry.

[0041] System 100 may provide a total residence time of the components of about 5 minutes to about 360 minutes, 5 minutes to about 180 minutes, 10 minutes to about 100 minutes, or more preferably about 30 minutes to about 70 minutes. For example, the system may provide a total residence time of about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 60 minutes, about 70 minutes, about 80 minutes, about 90 minutes, about 100 minutes, about 180 minutes, or about 360 minutes. The system may provide a total residence time of greater than about 90 minutes, greater than about 100 minutes, greater than about 180 minutes, or greater than about 360 minutes.

[0042] In some embodiments, one or more of the feeds may include a deuterated substance (e.g., deuterated BCD or deuterated propylene oxide). Use of the deuterated substance in one or more of the feeds may produce a deuterated HPBCD mixture.

[0043] In some embodiments, the crude HPBCD mixture collected in the collection tank 124 may be further purified through a purification process 200 as shown in FIGS. 26A - 26B. This purification process may be carried out as a batch process or a continuous process.

[0044] Prior to the start of the purification process 200, the HPBCD produced in the reactor 118 may be monitored at the junction 202 to determine the pH, concentration, and / or conductivity of the produced HPBCD, and / or other parameters of the HPBCD. If any parameter is determined to be outside a predetermined range, the HPBCD may be recycled through the reactor 118 before quenching in the collection tank 124.

[0045] Also, before the start of the purification process 200, the crude HPBCD mixture collected in the collection tank 124 may be monitored at the junction 204 to determine the pH, concentration, and / or conductivity of the crude HPBCD mixture, or other parameters of the mixture. If any parameter is determined to be outside the predetermined range, the HPBCD mixture may be recycled to the collection tank 124 before purification.

[0046] The purification process 200 of FIGS. 26A-26B begins with a first liquid filtration of the crude HPBCD mixture collected in the collection tank 124 using the filter 206. The filter 206 can be a liquid substance filter that can remove bulk solids and / or biological contaminants from the crude HPBCD mixture.

[0047] Next, the HPBCD mixture can be nanofiltrated using the membrane filter 208. This membrane can have a pore size of about 10 nm to about 1 nm, such as about 10 nm to about 5 nm, or about 5 nm to about 1 nm. In some embodiments, the membrane can have a pore size of about 10 nm, about 9 nm, about 8 nm, about 7 nm, about 6 nm, about 5 nm, about 4 nm, about 3 nm, about 2 nm, or about 1 nm. The membrane filter 208 can include regenerated cellulose, polyethersulfone, polyvinylidene fluoride, polypropylene, polyamide, polyethyleneimine, polyacrylonitrile, polyethylene, polytetrafluoroethylene, metal-organic frameworks, graphene, ceramics, composites, or other membrane materials known in the art and combinations thereof. Preferably, the membrane filter 208 includes regenerated cellulose or polyethersulfone.

[0048] In some embodiments, the filter 208 can include a flat sheet membrane to achieve nanofiltration. Flat sheet membranes for nanofiltration and methods of making and procuring flat sheet membranes are well known in the art. The flat sheet membrane is about 0.010 m 2 to about 0.500 m 2 , about 0.050 m 2 to about 0.100 m 2 , or about 0.010 m2 ~about 0.050 m 2 may have an area of. For example, the flat sheet membrane is about 0.010 m 2 , about 0.015 m 2 , about 0.020 m 2 , about 0.025 m 2 , about 0.030 m 2 , about 0.035 m 2 , about 0.040 m 2 , about 0.045 m 2 , or about 0.050 m 2 may have an area of. The flat sheet membrane is greater than 0.010 m 2 , greater than about 0.025 m 2 , greater than about 0.050 m 2 , greater than about 0.100 m 2 , greater than or about 0.500 m 2 may have an area greater than.

[0049] Nanofiltration can be achieved at a temperature of about 40°C to about 50°C, such as about 40°C to about 45°C, or about 45°C to about 50°C. In some embodiments, nanofiltration can be achieved at a temperature of about 40°C, about 41°C, about 42°C, about 43°C, about 44°C, about 45°C, about 46°C, about 47°C, about 48°C, about 49°C, or about 50°C.

[0050] Nanofiltration can be achieved at a pressure of about 1.5 MPa to about 2.0 MPa, such as about 1.5 MPa to about 1.75 MPa, or about 1.75 MPa to about 2.0 MPa. In some embodiments, nanofiltration can be achieved at a pressure of about 1.5 MPa, about 1.55 MPa, about 1.6 MPa, about 1.65 MPa, about 1.7 MPa, about 1.75 MPa, about 1.8 MPa, about 1.85 MPa, about 1.9 MPa, about 1.95 MPa, or about 2.0 MPa.

[0051] Alternatively, nanofiltration can be achieved at a pressure of about 0 psi to about 600 psi, about 50 psi to about 600 psi, about 100 psi to about 500 psi, about 200 psi to about 400 psi, or about 250 psi to about 350 psi. For example, purification can be carried out at a feed pressure of about 25 psi, about 50 psi, about 75 psi, about 100 psi, about 125 psi, about 150 psi, about 175 psi, about 200 psi, about 225 psi, about 250 psi, about 275 psi, about 300 psi, about 325 psi, about 350 psi, about 375 psi, about 400 psi, about 425 psi, about 450 psi, about 475 psi, or about 500 psi.

[0052] The nanofiltrated HPBCD mixture can have a conductivity of about 50 μS / cm or less, such as about 45 μS / cm or less, about 40 μS / cm or less, about 35 μS / cm or less, about 30 μS / cm or less, about 25 μS / cm or less, about 20 μS / cm or less, about 15 μS / cm or less, about 10 μS / cm or less, or about 5 μS / cm or less. Alternatively, the nanofiltrated HPBCD mixture can have a conductivity of about 0 μS / cm to about 50 μS / cm. For example, the nanofiltrated HPBCD mixture can have a conductivity of about 0 μS / cm to about 10 μS / cm, about 0 μS / cm to about 20 μS / cm, about 0 μS / cm to about 30 μS / cm, about 0 μS / cm to about 40 μS / cm, about 0 μS / cm to about 50 μS / cm, about 10 μS / cm to about 50 μS / cm, about 20 μS / cm to about 50 μS / cm, about 30 μS / cm to about 50 μS / cm, or about 40 μS / cm. In some embodiments, the nanofiltrated HPBCD mixture can have a conductivity of about 5 μS / cm, about 10 μS / cm, about 15 μS / cm, about 20 μS / cm, about 25 μS / cm, about 30 μS / cm, about 35 μS / cm, about 40 μS / cm, about 45 μS / cm, or about 50 μS / cm.

[0053] The nanofiltrated HPBCD mixture may have an impurity concentration of about 0.10 wt% or less. The impurities may include propylene glycol, propylene oxide, endotoxin, etc. For example, the nanofiltrated HPBCD mixture may have an impurity concentration of about 0.10 wt% or less, about 0.09 wt% or less, about 0.08 wt% or less, about 0.07 wt% or less, about 0.06 wt% or less, about 0.05 wt% or less, about 0.04 wt% or less, about 0.03 wt% or less, about 0.02 wt% or less, or about 0.01 wt% or less.

[0054] Before proceeding, the nanofiltrated HPBCD mixture may be monitored to determine the purity and conductivity of the nanofiltrated HPBCD mixture at the junction 210. If the purity and / or conductivity of the nanofiltrated HPBCD mixture is outside the predetermined range, the HPBCD mixture may be recirculated through the filter 208 at the junction 210 for further nanofiltration. Purified water may be added to the recirculated HPBCD mixture to facilitate subsequent nanofiltration.

[0055] After nanofiltration of the HPBCD mixture, the HPBCD mixture may be contacted with activated carbon in the container 214. The activated carbon may be useful for removing further impurities such as propylene oxide. The activated carbon may be prepared by first washing the activated carbon in the container 212 with purified water to remove salts. The activated carbon may be washed with purified water until the wash water has a conductivity of less than 10 μS / cm. Next, the activated carbon may be placed in the container 214 together with the nanofiltrated HPBCD mixture and may be agitated to ensure sufficient contact with the HPBCD mixture.

[0056] This contact may be carried out for a period of 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, 5 hours or more, 6 hours or more, 7 hours or more, 8 hours or more, 9 hours or more, or 10 hours or more.

[0057] This contact can be carried out at a temperature of about 15°C to about 30°C. For example, this contact can be carried out at a temperature of about 15°C to about 20°C, about 15°C to about 25°C, about 15°C to about 30°C, about 20°C to about 30°C, or about 25°C to about 30°C. In some examples, this contact can be carried out at a temperature of about 15°C, about 16°C, about 17°C, about 18°C, about 19°C, about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, or about 30°C.

[0058] After the contact, the HPBCD mixture may then be filtered through filter 216 to remove activated carbon from the mixture. Any filter capable of removing solid activated carbon from the liquid mixture may be used. Preferably, filter 216 includes a Nutsche filter.

[0059] Once the activated carbon has been filtered from the HPBCD mixture, the HPBCD mixture can have a propylene oxide concentration of less than 0.3 ppm. For example, the HPBCD mixture can have a propylene oxide concentration of about 0.2 ppm or less, about 0.1 ppm or less, about 0.09 ppm or less, about 0.08 ppm or less, about 0.07 ppm or less, about 0.06 ppm or less, about 0.05 ppm or less, about 0.04 ppm or less, about 0.03 ppm or less, about 0.02 ppm or less, or about 0.01 ppm or less. If the HPBCD mixture has a propylene oxide concentration of 0.3 ppm or more, the step of contacting the HPBCD mixture with activated carbon can be repeated until the propylene oxide concentration is less than 0.3 ppm.

[0060] Once the filtration of the activated carbon mixture is complete, the HPBCD mixture can have a conductivity of less than 90 μS / cm. For example, the HPBCD mixture can have a conductivity of about 80 μS / cm or less, about 70 μS / cm or less, about 60 μS / cm or less, about 50 μS / cm or less, about 40 μS / cm or less, about 30 μS / cm or less, about 20 μS / cm or less, or about 10 μS / cm or less. If the HPBCD mixture has a conductivity of 90 μS / cm or more, the step of nanofiltrating the HPBCD mixture can be repeated until the conductivity of the HPBCD mixture is less than 90 μS / cm.

[0061] Prior to proceeding, the HPBCD mixture may be monitored at the junction 218 to determine the propylene oxide concentration and / or conductivity of the HPBCD mixture. If the purity of the HPBCD mixture is outside a predetermined range, the HPBCD mixture may be recycled to the filter vessel 214 at the junction 218 for further purification. If the conductivity of the HPBCD mixture is outside a predetermined range, the HPBCD mixture may be recycled to the filter 208 at the junction 218 for further nanofiltration.

[0062] Once the activated carbon has been filtered and the HPBCD mixture has the desired purity and conductivity, the HPBCD mixture may be sterile filtered through the filter 220. Sterile filtration reduces the presence of bacteria and other microorganisms in the HPBCD mixture. Sterile filtration systems and methods are well known to those skilled in the art. The sterile filter preferably has a pore size of 0.22 μm or less. In some embodiments, the sterile filter may be a capsule filter. The sterile filter membrane may include polytetrafluoroethylene, polyethersulfone, polyvinylidene fluoride, nylon, polycarbonate, cellulose acetate, or other materials known in the art for sterile filtration and combinations thereof. The sterile filter preferably includes a polytetrafluoroethylene membrane.

[0063] The HPBCD mixture may then be filtered through a tangential flow filtration system 222. Tangential flow filtration systems and methods are well known to those skilled in the art. In some embodiments, the tangential flow filtration system 222 may include a membrane comprising polyethersulfone, polypropylene, polyurethane, regenerated cellulose, polyvinylidene fluoride, or other materials known in the art for tangential filtration membranes and combinations thereof. Preferably, the membrane includes polyethersulfone.

[0064] After the HPBCD mixture has been filtered through the tangential flow filtration system 222, the HPBCD mixture may be dried in a dryer 224. Preferably, the HPBCD mixture is spray dried.

[0065] In embodiments where the HPBCD mixture is spray dried, the inlet temperature of the spray dryer can be from about 180°C to about 220°C. For example, the inlet temperature of the spray dryer can be from about 180°C to about 190°C, from about 180°C to about 200°C, from about 180°C to about 210°C, from about 180°C to about 220°C, from about 190°C to about 200°C, from about 190°C to about 210°C, from about 190°C to about 220°C, from about 200°C to about 210°C, from about 200°C to about 220°C, or from about 210°C to about 220°C. The outlet temperature of the spray dryer can be from about 100°C to about 120°C. For example, the outlet temperature of the spray dryer can be from about 100°C to about 105°C, from about 100°C to about 110°C, from about 100°C to about 115°C, from about 100°C to about 120°C, from about 105°C to about 110°C, from about 105°C to about 115°C, from about 105°C to about 120°C, from about 110°C to about 115°C, from about 110°C to about 120°C, or from about 115°C to about 120°C.

[0066] Furthermore, provided herein is a method for producing an HPBCD mixture. The method can be achieved using any of the systems described above. The method includes (a) contacting an HPBCD mixture with at least two solvents, wherein the HPBCD mixture includes highly substituted HPBCD and lowly substituted HPBCD, (b) dissolving the highly substituted HPBCD in one of the solvents, and (c) removing the lowly substituted HPBCD by precipitation. The at least two solvents can include ethanol and acetone. The highly substituted HPBCD can have an average degree of substitution of about 6.0 or greater, about 6.5 or greater, about 7.0 or greater, about 7.5 or greater, about 8.0 or greater, about 8.5 or greater, about 9.0 or greater, or about 9.5 or greater. The lowly substituted HPBCD can have an average degree of substitution of less than about 7.5, less than about 7.0, less than about 6.5, less than about 6.0, less than about 5.5, or less than about 5.0.

[0067] Alternatively, the method may include (a) contacting an HPBCD mixture with at least two solvents, wherein the HPBCD mixture comprises highly substituted HPBCD; (b) dissolving the highly substituted HPBCD in one of the solvents to form a mother liquor; and (c) filtering off the mother liquor. The method may further include crystallizing or lyophilizing the mother liquor to obtain a solid. The solid may be analyzed by MALDI-TOF to determine the degree of substitution.

[0068] In some embodiments, deuterated reactants (e.g., deuterated BCD or deuterated propylene oxide) can be used to obtain deuterated products such as deuterated HPBCD.

[0069] System 100 may further comprise a purification system for purifying HPBCD. The purification system may include absorption chromatography alumina, solvent precipitation, or a combination thereof.

[0070] Furthermore, provided herein is a method for oligomeric substitution by methanolysis of an HPBCD mixture. The method generally includes mixing HPBCD and methanol, stirring until the HPBCD dissolves, adding an acid to the mixture, heating the mixture to at least about 50 to about 90 °C, stirring the mixture and maintaining the heat for at least about 24 hours, neutralizing the mixture with a base, and filtering the mixture. In some embodiments, the HPBCD can be a racemic mixture of HPBCD, and in other embodiments, the HPBCD can be enantiopure HPBCD.

[0071] Methanol can be added in an amount of about 100 to about 300 molar equivalents of HPBCD. For example, methanol can be added in an amount of about 50, about 75, about 100, about 125, about 150, about 175, about 200, about 225, about 250, about 275, about 300, about 350, or about 400 molar equivalents of HPBCD. In some embodiments, methanol can include deuterated methanol.

[0072] The acids added to the mixture can include hydrochloric acid, sulfuric acid, lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, malic acid, fumaric acid, tartaric acid, or combinations thereof. In an exemplary embodiment, the acid includes sulfuric acid.

[0073] The heat of the mixture can be maintained for at least 24 hours. For example, the heat can be maintained for 24 hours, 30 hours, 36 hours, 42 hours, 48 hours, or longer than 48 hours.

[0074] The bases used to neutralize the mixture can include sodium hydroxide, potassium hydroxide, lithium hydroxide, magnesium hydroxide, calcium hydroxide, or combinations thereof. In an exemplary embodiment, the base is sodium hydroxide.

[0075] The mixture can be filtered by filtration methods widely known in the art. In a preferred embodiment, the mixture is filtered by the nanofiltration method described below.

[0076] Further provided herein is a method for purifying an HPBCD mixture, comprising purifying the HPBCD mixture by nanofiltration, collecting at least a total of five volumes of diafiltration amount of nanofiltration permeate, and lyophilizing the resulting residue to obtain solid hydroxypropyl-β-cyclodextrin. In some embodiments, different volumes of diafiltration amount can produce different HPBCD mixtures.

[0077] The HPBCD mixture is purified by nanofiltration. Purification by nanofiltration can include a flat sheet membrane for achieving nanofiltration. Flat sheet membranes for nanofiltration and methods for fabricating and procuring flat sheet membranes are widely known in the art. The flat sheet membrane is about 0.010 m 2 ~ about 0.500 m 2 、 about 0.050 m 2 ~ about 0.100 m 2 、 or about 0.010 m 2 ~ about 0.050 m 2may have an area. For example, the flat sheet membrane may have an area of about 0.010 m 2 , about 0.015 m 2 , about 0.020 m 2 , about 0.025 m 2 , about 0.030 m 2 , about 0.035 m 2 , about 0.040 m 2 , about 0.045 m 2 , or about 0.050 m 2 . The flat sheet membrane may have an area greater than 0.010 m 2 , greater than about 0.025 m 2 , greater than about 0.050 m 2 , greater than about 0.100 m 2 , or greater than about 0.500 m 2 .

[0078] Purification and / or feed may be carried out at a feed pressure of about 0 psi to about 600 psi, about 50 psi to about 600 psi, about 100 psi to about 500 psi, about 200 psi to about 400 psi, about 250 psi to about 350 psi. For example, purification may be carried out at a feed pressure of about 25 psi, about 50 psi, about 75 psi, about 100 psi, about 125 psi, about 150 psi, about 175 psi, about 200 psi, about 225 psi, about 250 psi, about 275 psi, about 300 psi, about 325 psi, about 350 psi, about 375 psi, about 400 psi, about 425 psi, about 450 psi, about 475 psi, or about 500 psi.

[0079] The collection of the nanofiltration permeate can be achieved for at least one volume of diafiltration, at least two volumes of diafiltration, at least three volumes of diafiltration, at least four volumes of diafiltration, or at least five volumes of diafiltration in total. For example, the nanofiltration permeate can be collected for at least five volumes of diafiltration, at least six volumes of diafiltration, at least seven volumes of diafiltration, at least eight volumes of diafiltration, at least nine volumes of diafiltration, or at least ten volumes of diafiltration in total. In some embodiments, the nanofiltration permeate can be collected for a diafiltration volume exceeding ten volumes.

[0080] The method may further include analyzing the propylene glycol content of the resulting residue. Methods for analyzing the propylene glycol content of a composition are well known in the art and may include mass spectrometry, high performance liquid chromatography, gas chromatography, etc.

[0081] Further provided herein is a method for purifying an HPBCD mixture, comprising purifying the HPBCD mixture by nanofiltration, collecting a nanofiltration permeate with a diafiltration volume of at least five volumes in total, and analyzing the propylene glycol content of the resulting residue.

[0082] Further provided herein is a composition comprising a methylated 2-hydroxypropyl-β-cyclodextrin mixture having an average degree of substitution of from about 6.5 to about 9.5 and a methylated glucose having from 0 to about 5 2-hydroxypropyl groups. For example, the methylated 2-hydroxypropyl-β-cyclodextrin mixture can have an average degree of substitution of about 6.5, about 7.0, about 7.5, about 8.0, about 8.5, about 9.0, or about 9.5. The methylated 2-hydroxypropyl-β-cyclodextrin mixture can have an average degree of substitution of from about 6.5 to about 9.5, from about 6.5 to about 9.0, from about 6.8 to about 9.5, from about 6.8 to about 9.0, from about 7.0 to about 9.5, from about 7.0 to about 9.0, from about 7.2 to about 9.5, from about 7.2 to about 9.0, from about 7.5 to about 9.5, from about 7.5 to about 9.0, from about 7.8 to about 9.5, from about 7.8 to about 9.0, from about 8.0 to about 9.5, from about 8.0 to about 9.0, from about 8.2 to about 9.5, from about 8.2 to about 9.0, from about 8.5 to about 9.5, from about 8.5 to about 9.0, from about 8.8 to about 9.5, or from about 8.8 to about 9.0. In an exemplary embodiment, the composition has a mass spectrum as shown in FIG. 25.

[0083] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 0.3% unsubstituted beta-cyclodextrin ("DS-0") or less than 1% beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The mixture may comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1. The amount of DS-0 or DS-1 can be determined by the peak height of an electrospray MS spectrum.

[0084] The mixture can have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture can have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture can have an average degree of substitution ("DS a ") in the range of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7. For example, the mixture can have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0085] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0086] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0087] The total amount of other unspecified impurities in the composition may be 0.05% or less. For example, the total amount of unspecified impurities in the composition may be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0088] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient can be an adult patient or a pediatric patient. The composition may further contain a pharmaceutically acceptable diluent.

[0089] The composition can solubilize lipids in an aqueous medium. The lipids can include non-esterified or esterified cholesterol. The composition may be provided as a solution with a solution concentration of 20 w / v% of a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions. The composition can have an affinity for non-esterified cholesterol. The solubilization can be determined by UV spectroscopy or HPLC.

[0090] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0091] The composition may have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition may have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0092] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 2.5% beta-cyclodextrin substituted with one hydroxypropyl group (“DS-1”), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The mixture may comprise less than 2.5%, less than 2.4%, less than 2.3%, less than 2.2%, less than 2.1%, less than 2.0%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1.0%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% DS-1. The amount of DS-1 can be determined by the peak height of the electrospray MS spectrum.

[0093] The composition may comprise 1% or less unsubstituted beta-cyclodextrin (“DS-0”). For example, the composition may comprise 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, 0.1% or less, 0.09% or less, 0.08% or less, 0.07% or less, 0.06% or less, 0.05% or less, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less DS-0. The amount of DS-0 can be determined by the peak height of the electrospray MS spectrum.

[0094] The mixture may have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture may have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture has an average degree of substitution (“DS” of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7a It may have ")". For example, the mixture may have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0095] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0096] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0097] The total amount of other unspecified impurities in the composition can be 0.05% or less. For example, the total amount of unspecified impurities in the composition can be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0098] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient can be an adult patient or a pediatric patient. The composition may further contain a pharmaceutically acceptable diluent.

[0099] The composition can solubilize lipids in an aqueous medium. The lipids can include non-esterified or esterified cholesterol. The composition may be provided as a solution having a concentration of 20 w / v% in the solution of the composition. The composition can have an affinity for non-esterified cholesterol. The solubilization can be determined by UV spectroscopy or HPLC.

[0100] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0101] The composition can have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition can have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0102] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin (“DS-0”) and beta-cyclodextrin substituted with one hydroxypropyl group (“DS-1”), and the mixture comprises from 5% to 25% beta-cyclodextrin substituted with six hydroxypropyl groups (“DS-6”).

[0103] The mixture may comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0104] The mixture may comprise at least 8% beta-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”). The mixture may comprise at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, or at least 25% of DS-6. Alternatively, the mixture may comprise from about 8% to about 9%, from about 8% to about 10%, from about 8% to about 11%, from about 8% to about 12%, from about 8% to about 13%, from about 8% to about 14%, from about 8% to about 15%, from about 8% to about 16%, from about 8% to about 17%, from about 8% to about 18%, from about 8% to about 19%, from about 8% to about 20%, from about 8% to about 21%, from about 8% to about 22%, from about 8% to about 23%, from about 8% to about 24%, or from about 8% to about 25%. Alternatively, the mixture may comprise 15% or less, 14% or less, 13% or less, 12% or less, 11% or less, 10% or less, 9% or less, or 8% or less of DS-6.

[0105] The amount of DS-0, DS-1, or DS-6 can be determined by the peak height of the electrospray MS spectrum.

[0106] The mixture may have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture may have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture may have an average degree of substitution (“DS a ”) in the range of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7. For example, the mixture may have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0107] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0108] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0109] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0110] The total amount of other unspecified impurities in the composition may be 0.05% or less. For example, the total amount of unspecified impurities in the composition may be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0111] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient may be an adult patient or a pediatric patient. The composition may further comprise a pharmaceutically acceptable diluent.

[0112] The composition may solubilize lipids in an aqueous medium. The lipids may include non-esterified or esterified cholesterol. The composition may be provided as a solution having a concentration of 20 w / v% in the solution of the composition. The composition may have an affinity for non-esterified cholesterol. The solubilization may be determined by UV spectroscopy or HPLC.

[0113] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0114] The composition may have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition may have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0115] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises from 1% to 10% beta-cyclodextrin substituted with seven hydroxypropyl groups ("DS-7").

[0116] The mixture may comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0117] The mixture may comprise from about 1% to about 10% of DS-7. For example, the mixture may comprise from about 1% to about 2%, from about 1% to about 3%, from about 1% to about 4%, from about 1% to about 5%, from about 1% to about 6%, from about 1% to about 7%, from about 1% to about 8%, from about 1% to about 9%, from about 2% to about 10%, from about 3% to about 10%, from about 4% to about 10%, from about 5% to about 10%, from about 6% to about 10%, from about 7% to about 10%, from about 8% to about 10%, from about 9% to about 10%, from about 2% to about 9%, from about 3% to about 8%, from about 4% to about 7%, or from about 5% to about 6% of DS-7. The mixture may comprise about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or about 10% of DS-7. Alternatively, the composition may have from about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less of DS-7.

[0118] The amount of DS-0, DS-1, or DS-7 can be determined by the peak height of the electrospray MS spectrum.

[0119] The mixture can have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture can have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture can have an average degree of substitution (''DS a '') in the range of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7. For example, the mixture can have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0120] The composition can contain propylene glycol at 0.01% or less. For example, the composition can contain propylene glycol at 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0121] The composition can contain propylene glycol at 0.01% or less. For example, the composition can contain propylene glycol at 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0122] The composition may contain propylene oxide at 1 ppm or less, 0.9 ppm or less, 0.8 ppm or less, 0.7 ppm or less, 0.6 ppm or less, 0.5 ppm or less, 0.4 ppm or less, 0.3 ppm or less, 0.2 ppm or less, or 0.1 ppm or less. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0123] The total amount of other unspecified impurities in the composition can be 0.05% or less. For example, the total amount of unspecified impurities in the composition can be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0124] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient can be an adult patient or a pediatric patient. The composition may further contain a pharmaceutically acceptable diluent.

[0125] The composition can solubilize lipids in an aqueous medium. The lipid can include non-esterified or esterified cholesterol. The composition may be provided as a solution with a concentration of 20 w / v% in the solution of the composition. The composition can have an affinity for non-esterified cholesterol. The solubilization can be determined by UV spectroscopy or HPLC.

[0126] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0127] The composition can have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition can have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0128] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 50% or less of beta-cyclodextrin substituted with four hydroxypropyl groups ("DS-4").

[0129] The mixture may comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0130] The mixture may contain up to 25% of beta-cyclodextrin substituted with 4 hydroxypropyl groups ("DS-4"). The mixture may contain at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, or at least 25% of DS-4. Alternatively, the mixture may contain up to 25%, up to 30%, up to 35%, up to 40%, up to 45%, or up to 50% of DS-4. The mixture may contain from about 5% to about 50%, from about 5% to about 10%, from about 5% to about 20%, from about 5% to about 30%, from about 5% to about 40%, from about 10% to about 50%, from about 20% to about 50%, from about 30% to about 50%, from about 40% to about 50%, from about 10% to about 40%, or from about 20% to about 30% of DS-4.

[0131] The amount of DS-0, DS-1, or DS-4 can be determined by the peak height of the electrospray MS spectrum.

[0132] The mixture may have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture may have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture may have an average degree of substitution ("DS a ") in the range of about 3 to about 7, from about 4 to about 7, from about 5 to about 7, or from about 6 to about 7. For example, the mixture may have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0133] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0134] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0135] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0136] The total amount of other unspecified impurities in the composition may be 0.05% or less. For example, the total amount of unspecified impurities in the composition may be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0137] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient may be an adult patient or a pediatric patient. The composition may further comprise a pharmaceutically acceptable diluent.

[0138] The composition may solubilize lipids in an aqueous medium. The lipids may include non-esterified or esterified cholesterol. The composition may be provided as a solution having a concentration of 20 w / v% in the solution of the composition. The composition may have an affinity for non-esterified cholesterol. Solubilization may be determined by UV spectroscopy or HPLC.

[0139] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0140] The composition may have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition may have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0141] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin (“DS-0”) and beta-cyclodextrin substituted with one hydroxypropyl group (“DS-1”), and the mixture comprises 50% or less of beta-cyclodextrin substituted with five hydroxypropyl groups (“DS-5”).

[0142] The mixture can comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture can comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture can comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0143] The mixture may contain up to 25% of beta-cyclodextrin substituted with 5 hydroxypropyl groups ("DS-5"). The mixture may contain at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, or at least 25% of DS-5. Alternatively, the mixture may contain up to 25%, up to 30%, up to 35%, up to 40%, up to 45%, or up to 50% of DS-5. The mixture may contain from about 5% to about 50%, from about 5% to about 10%, from about 5% to about 20%, from about 5% to about 30%, from about 5% to about 40%, from about 10% to about 50%, from about 20% to about 50%, from about 30% to about 50%, from about 40% to about 50%, from about 10% to about 40%, or from about 20% to about 30% of DS-5.

[0144] The amount of DS-0, DS-1, or DS-5 can be determined by the peak height of the electrospray MS spectrum.

[0145] The mixture may have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture may have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture may have an average degree of substitution ("DS a ") in the range of about 3 to about 7, from about 4 to about 7, from about 5 to about 7, or from about 6 to about 7. For example, the mixture may have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0146] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0147] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0148] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0149] The total amount of other unspecified impurities in the composition can be 0.05% or less. For example, the total amount of unspecified impurities in the composition can be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0150] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient may be an adult patient or a pediatric patient. The composition may further comprise a pharmaceutically acceptable diluent.

[0151] The composition may solubilize lipids in an aqueous medium. The lipid may comprise non-esterified or esterified cholesterol. The composition may be provided as a solution having a concentration in the composition solution of 20 w / v%. The composition may have an affinity for non-esterified cholesterol. The solubilization may be determined by UV spectroscopy or HPLC.

[0152] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0153] The composition can have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition can have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0154] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and at least 70% of the beta-cyclodextrin has a DS a within ±1σ, where σ is the standard deviation, the composition.

[0155] At least 70% of the beta-cyclodextrin has a DS a within ±1σ, where σ is the standard deviation. In some embodiments, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the beta-cyclodextrin has a DS a within ±1σ.

[0156] The mixture may comprise less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may comprise less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0157] The amount of DS-0 or DS-1 can be determined by the peak height of the electrospray MS spectrum.

[0158] The mixture can have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture can have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture can have an average degree of substitution (「DS a 」) in the range of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7. For example, the mixture can have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7.

[0159] The composition can contain 0.01% or less of propylene glycol. For example, the composition can contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0160] The composition can contain 0.01% or less of propylene glycol. For example, the composition can contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0161] The composition can contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0162] The total amount of other unspecified impurities in the composition can be 0.05% or less. For example, the total amount of unspecified impurities in the composition can be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0163] The composition can be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient can be an adult patient or a pediatric patient. The composition can further contain a pharmaceutically acceptable diluent.

[0164] The composition can solubilize lipids in an aqueous medium. The lipid can contain non-esterified or esterified cholesterol. The composition may be provided as a solution with a concentration of 20 w / v% in the solution of the composition. The composition can have an affinity for non-esterified cholesterol. The solubilization can be determined by UV spectroscopy or HPLC.

[0165] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0166] The composition may have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition may have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0167] Further provided herein is a composition produced using one or more of the systems and / or methods provided herein, the composition comprising a mixture of β-cyclodextrin molecules, the mixture of β-cyclodextrin molecules comprising β-cyclodextrin substituted with 0 hydroxypropyl groups (“DS-0,” also referred to as “unsubstituted”), β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). The degree of substitution of the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. As relevant to this document, the number of hydroxypropyl groups per anhydroglucose unit in the mixture of beta-cyclodextrin is the “molar substitution” or “MS” and is determined according to the procedure described in the USP monograph for Hydroxypropyl Betadex (USP NF 2015) (“USP Hydroxypropyl Betadex monograph”), which is incorporated herein by reference in its entirety. In the present disclosure, “average molar substitution” or “MS aThe term "___" is used synonymously with the term "MS" as used in the USP monograph on hydroxypropyl beta dextrin, and the term "glucose unit" is used as a synonym for the term "anhydrous glucose unit" as used in the USP monograph on hydroxypropyl beta dextrin. Further, as related to this book, the "average number of hydroxypropyl groups per beta-cyclodextrin" is known as the "degree of substitution", "average DS", or "DS a ", and refers to the total number of hydroxypropyl groups in a population of beta-cyclodextrins divided by the number of beta-cyclodextrin molecules. In an illustrative example, an equimolar mixture of beta-cyclodextrin containing glucose units each substituted with one hydroxypropyl group and beta-cyclodextrin containing glucose units each substituted with two hydroxypropyl groups has a DS a = 10.5 (average of equimolar amounts of beta-cyclodextrin with DS = 7 and DS = 14). In another illustrative example, a mixture of 33.3% beta-cyclodextrin in which only 1 out of 7 glucose units is substituted with a hydroxypropyl group (i.e., DS = 1) and 66.7% beta-cyclodextrin containing glucose units each substituted with one hydroxypropyl group (i.e., DS = 7) has a DS a = 5.0. DS a is determined by multiplying MS by 7. Further, as related to this book, the "degree of substitution" or "DS" refers to the total number of hydroxypropyl groups directly or indirectly substituted on the beta-cyclodextrin molecule. For example, a beta-cyclodextrin molecule containing glucose units each substituted with one hydroxypropyl group has a DS = 7. In another example, a beta-cyclodextrin molecule in which only 1 out of 7 glucose units is substituted with a hydroxypropyl group and that hydroxypropyl group itself is substituted with another hydroxypropyl group (e.g., one occurrence of HP containing two hydroxypropyl groups in one beta-cyclodextrin) has a DS = 2. As used herein, DS ais used synonymously with the term "degree of substitution" as defined in the USP monograph for hydroxypropyl beta dextrin.

[0168] In certain embodiments, the pharmaceutical composition of the present disclosure comprises, as a pharmaceutically active ingredient, a mixture of unsubstituted beta-cyclodextrin molecules and beta-cyclodextrin molecules substituted at one or more hydroxyl positions with hydroxypropyl groups, the mixture having an average number of hydroxypropyl groups per beta-cyclodextrin molecule (DS a ) of from about 3 to about 7.

[0169] In some embodiments, DS a is from about 3 to about 5, such as from about 3 to about 4. In some embodiments, DS a is 3.3 ± 0.3, 3.5 ± 0.3, or 3.7 ± 0.3. In other embodiments, DS a is 3.2 ± 0.2, 3.3 ± 0.2, 3.4 ± 0.2, 3.5 ± 0.2, 3.6 ± 0.2, 3.7 ± 0.2, or 3.8 ± 0.2. In other embodiments, DS a is 3.1 ± 0.1, 3.2 ± 0.1, 3.3 ± 0.1, 3.4 ± 0.1, ±0.1, 3.6 ± 0.1, 3.7 ± 0.1, 3.8 ± 0.1, or 3.9 ± 0.1.

[0170] In some embodiments, DS a is from about 3.5 to about 5.5, such as from about 3.5 to about 4.5. In some embodiments, DS a is 3.8 ± 0.3, 4.0 ± 0.3, or 4.2 ± 0.3. In other embodiments, DS a is 3.7 ± 0.2, 3.8 ± 0.2, 3.9 ± 0.2, 4.0 ± 0.2, 4.1 ± 0.2, 4.2 ± 0.2, or 4.3 ± 0.2. In other embodiments, DS a is 3.6 ± 0.1, 3.7 ± 0.1, 3.8 ± 0.1, 3.9 ± 0.1, 4.0 ± 0.1, 4.1 ± 0.1, 4.2 ± 0.1, 4.3 ± 0.1, or 4.4 ± 0.1.

[0171] In some embodiments, DS a is from about 4 to about 6, such as from about 4 to about 5. In some embodiments, DS a is 4.3 ± 0.3, 4.5 ± 0.3, or 4.7 ± 0.3. In other embodiments, DS a is 4.2 ± 0.2, 4.3 ± 0.2, 4.4 ± 0.2, 4.5 ± 0.2, 4.6 ± 0.2, 4.7 ± 0.2, or 4.8 ± 0.2. In other embodiments, DS a is 4.1 ± 0.1, 4.2 ± 0.1, 4.3 ± 0.1, 4.4 ± 0.1, 4.5 ± 0.1, 4.6 ± 0.1, 4.7 ± 0.1, 4.8 ± 0.1, or 4.9 ± 0.1.

[0172] In some embodiments, DS a is from about 4.5 to about 6.5, such as from about 4.5 to about 5.5. In some embodiments, DS a is 4.8 ± 0.3, 5.0 ± 0.3, or 5.2 ± 0.3. In other embodiments, DS a is 4.7 ± 0.2, 4.8 ± 0.2, 4.9 ± 0.2, 5.0 ± 0.2, 5.1 ± 0.2, 5.2 ± 0.2, or 5.3 ± 0.2. In other embodiments, DS a is 4.6 ± 0.1, 4.7 ± 0.1, 4.8 ± 0.1, 4.9 ± 0.1, 5.0 ± 0.1, 5.1 ± 0.1, 5.2 ± 0.1, 5.3 ± 0.1, or 5.4 ± 0.1.

[0173] In some embodiments, DS a is from about 5 to about 7, such as from about 5 to about 6. In some embodiments, DS a is 5.3 ± 0.3, 5.5 ± 0.3, or 5.7 ± 0.3. In other embodiments, DS a is 5.2 ± 0.2, 5.3 ± 0.2, 5.4 ± 0.2, 5.5 ± 0.2, 5.6 ± 0.2, 5.7 ± 0.2, or 5.8 ± 0.2. In other embodiments, DS a is 5.1 ± 0.1, 5.2 ± 0.1, 5.3 ± 0.1, 5.4 ± 0.1, 5.5 ± 0.1, 5.6 ± 0.1, 5.7 ± 0.1, 5.8 ± 0.1, or 5.9 ± 0.1.

[0174] In some embodiments, DS a is from about 5.5 to about 6.5. In some embodiments, DS a is 5.8 ± 0.3, 6.0 ± 0.3, or 6.2 ± 0.3. In other embodiments, DS a is 5.7 ± 0.2, 5.8 ± 0.2, 5.9 ± 0.2, 6.0 ± 0.2, 6.1 ± 0.2, 6.2 ± 0.2, or 6.3 ± 0.2. In other embodiments, DS a is 5.6 ± 0.1, 5.7 ± 0.1, 5.8 ± 0.1, 5.9 ± 0.1, 6.0 ± 0.1, 6.1 ± 0.1, 6.2 ± 0.1, 6.3 ± 0.1, or 6.4 ± 0.1.

[0175] In some embodiments, DS a is from about 6 to about 7. In some embodiments, DS a is 6.3 ± 0.3, 6.5 ± 0.3, or 6.7 ± 0.3. In other embodiments, DS a is 6.2 ± 0.2, 6.3 ± 0.2, 6.4 ± 0.2, 6.5 ± 0.2, 6.6 ± 0.2, 6.7 ± 0.2, or 6.8 ± 0.2. In other embodiments, DS a is 6.1 ± 0.1, 6.2 ± 0.1, 6.3 ± 0.1, 6.4 ± 0.1, 6.5 ± 0.1, 6.6 ± 0.1, 6.7 ± 0.1, 6.8 ± 0.1, or 6.9 ± 0.1.

[0176] In some embodiments, DS a is about 4.1 ± 15%, about 4.2 ± 15%, about 4.3 ± 15%, about 4.4 ± 15%, or about 4.5 ± 15%, for example about 4.1 ± 10%, about 4.2 ± 10%, about 4.3 ± 10%, about 4.4 ± 10%, or about 4.5 ± 10%, for example about 4.1 ± 5%, about 4.2 ± 5%, about 4.3 ± 5%, about 4.4 ± 5%, or about 4.5 ± 5%. For example, in certain embodiments, DS ais about 4.31±10%, about 4.32±10%, about 4.33±10%, about 4.34±10%, about 4.35±10%, about 4.36±10%, or about 4.37±10%, for example about 4.31±5%, about 4.32±5%, about 4.33±5%, about 4.34±5%, about 4.35±5%, about 4.36±5%, or about 4.37±5%. In certain embodiments, DS a is about 4.34±10%, for example about 4.34±5%.

[0177] In some embodiments, DS a is about 4.3±15%, about 4.4±15%, about 4.5±15%, about 4.6±15%, or about 4.7±15%, for example about 4.3±10%, about 4.4±10%, about 4.5±10%, about 4.6±10%, or about 4.7±10%, for example about 4.3±5%, about 4.4±5%, about 4.5±5%, about 4.6±5%, or about 4.7±5%. For example, in certain embodiments, DS a is about 4.47±10%, about 4.48±10%, about 4.49±10%, about 4.50±10%, about 4.51±10%, about 4.52±10%, or about 4.53±10%, for example about 4.47±5%, about 4.48±5%, about 4.49±5%, about 4.50±5%, about 4.51±5%, about 4.52±5%, or about 4.53±5%. In certain embodiments, DS a is about 4.50±10%, for example about 4.50±5%.

[0178] In some embodiments, DS a is about 6.1±15%, about 6.2±15%, about 6.3±15%, about 6.4±15%, or about 6.5±15%, for example about 6.1±10%, about 6.2±10%, about 6.3±10%, about 6.4±10%, or about 6.5±10%, for example about 6.1±5%, about 6.2±5%, about 6.3±5%, about 6.4±5%, or about 6.5±5%. For example, in certain embodiments, DS ais about 6.34±10%, about 6.35±10%, about 6.36±10%, about 6.37±10%, about 6.38±10%, about 6.39±10%, or about 6.40±10%, for example about 6.34±5%, about 6.35±5%, about 6.36±5%, about 6.37±5%, about 6.38±5%, about 6.39±5%, or about 6.40±5%. In certain embodiments, DS a is about 6.37±10%, for example about 6.37±5%.

[0179] In some embodiments, DS a is about 6.3±15%, about 6.4±15%, about 6.5±15%, about 6.6±15%, or about 6.7±15%, for example about 6.3±10%, about 6.4±10%, about 6.5±10%, about 6.6±10%, or about 6.7±10%, for example about 6.3±5%, about 6.4±5%, about 6.5±5%, about 6.6±5%, or about 6.7±5%. For example, in certain embodiments, DS a is about 6.50±10%, about 6.51±10%, about 6.52±10%, about 6.53±10%, about 6.54±10%, about 6.55±10%, or about 6.56±10%, for example about 6.50±5%, about 6.51±5%, about 6.52±5%, about 6.53±5%, about 6.54±5%, about 6.55±5%, or about 6.56±5%. In certain embodiments, DS a is about 6.53±10%, for example about 6.53±5%.

[0180] The distribution of the degree of substitution in a mixture of unsubstituted beta-cyclodextrin molecules and beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions can vary. For example, an equimolar mixture of beta-cyclodextrin containing glucose units each substituted with one hydroxypropyl group and beta-cyclodextrin containing glucose units each substituted with two hydroxypropyl groups has a DS a = 10.5 (average of equimolar amounts of beta-cyclodextrin with DS = 7 and DS = 14). DS a= 10.5, but in this example, beta-cyclodextrin having DS = 10 or DS = 11 is not present in the mixture. In other cases, most of the beta-cyclodextrin in the mixture of beta-cyclodextrin has a DS close to this DS a having a DS close thereto.

[0181] In some embodiments of the present disclosure, at least about 50%, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% of the beta-cyclodextrin in the mixture has a DS a within DS ± Xσ, where σ is the standard deviation and X is 1, 2, or 3. For example, in some embodiments, at least about 50%, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% of the beta-cyclodextrin in the mixture has a DS a within DS ± 1σ. In some embodiments, at least about 70% of the beta-cyclodextrin has a DS a within DS ± 1σ. In some embodiments, at least about 90% of the beta-cyclodextrin has a DS a within DS ± 1σ. In some embodiments, at least about 95% of the beta-cyclodextrin has a DS a within DS ± 1σ.

[0182] In some embodiments, at least about 50%, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% of the beta-cyclodextrin in the mixture has a DS a within DS ± 2σ. In some embodiments, at least about 70% of the beta-cyclodextrin has a DS a within DS ± 2σ. In some embodiments, at least about 90% of the beta-cyclodextrin has a DS a within DS ± 2σ. In some embodiments, at least about 95% of the beta-cyclodextrin has a DSa It has a DS within ±2σ.

[0183] In some embodiments, at least about 50% of the beta-cyclodextrin in the mixture, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±3σ. In some embodiments, at least about 70% of the beta-cyclodextrin has a DS within DSa ±3σ. In some embodiments, at least about 90% of the beta-cyclodextrin has a DS a within ±3σ. In some embodiments, at least about 95% of the beta-cyclodextrin has a DS a within ±3σ.

[0184] In some embodiments, at least about 50% of the beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±1. In some embodiments, at least about 70% of the beta-cyclodextrin has a DS a within ±1. In some embodiments, at least about 90% of the beta-cyclodextrin has a DS a within ±1. In some embodiments, at least about 95% of the beta-cyclodextrin has a DS a within ±1.

[0185] In some embodiments, at least about 50% of the beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±0.8. In some embodiments, at least about 70% of the beta-cyclodextrin has a DS aIt has a DS within ±0.8. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a It has a DS within ±0.8. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a It has a DS within ±0.8.

[0186] In some embodiments, at least about 50% of beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a It has a DS within ±0.6. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a It has a DS within ±0.6. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a It has a DS within ±0.6. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a It has a DS within ±0.6.

[0187] In some embodiments, at least about 50% of beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a It has a DS within ±0.5. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a It has a DS within ±0.5. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a It has a DS within ±0.5. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a It has a DS within ±0.5.

[0188] In some embodiments, at least about 50% of beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±0.4 of the DS. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a within ±0.4 of the DS. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a within ±0.4 of the DS. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a within ±0.4 of the DS.

[0189] In some embodiments, at least about 50% of beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±0.3 of the DS. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a within ±0.3 of the DS. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a within ±0.3 of the DS. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a within ±0.3 of the DS.

[0190] In some embodiments, at least about 50% of beta-cyclodextrin, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% has a DS a within ±0.2 of the DS. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a within ±0.2 of the DS. In some embodiments, at least about 90% of beta-cyclodextrin has a DS aIt has a DS within ±0.2. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a It has a DS within ±0.2.

[0191] In some embodiments, at least about 50%, such as at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 97% of beta-cyclodextrin has a DS a It has a DS within ±0.1. In some embodiments, at least about 70% of beta-cyclodextrin has a DS a It has a DS within ±0.1. In some embodiments, at least about 90% of beta-cyclodextrin has a DS a It has a DS within ±0.1. In some embodiments, at least about 95% of beta-cyclodextrin has a DS a It has a DS within ±0.1.

[0192] In some embodiments, the MS ranges from 0.40 to 0.80, such as 0.41 to 0.79, 0.42 to 0.78, 0.43 to 0.77, 0.44 to 0.76, 0.45 to 0.75, 0.46 to 0.74, 0.47 to 0.73, 0.48 to 0.72, 0.49 to 0.71, 0.50 to 0.70, 0.51 to 0.69, 0.52 to 0.68, 0.53 to 0.67, 0.54 to 0.66, 0.55 to 0.65, 0.56 to 0.64, 0.57 to 0.63, 0.58 to 0.62, or 0.59 to 0.61.

[0193] In certain embodiments, the MS is about 0.40, about 0.41, about 0.42, about 0.43, about 0.44, about 0.45, about 0.46, about 0.47, about 0.48, about 0.49, about 0.50, about 0.51, about 0.52, about 0.53, about 0.54, about 0.55, about 0.56, about 0.57, about 0.58, about 0.59, about 0.60, about 0.61, about 0.62, about 0.63, about 0.64, about 0.65, about 0.66, about 0.67, about 0.68, about 0.69, about 0.70, about 0.71, about 0.72, about 0.73, about 0.74, about 0.75, about 0.76, about 0.77, about 0.78, about 0.79, or about 0.80.

[0194] In certain embodiments, the MS is about 0.571 - 0.686 (DS a about 4.0 - about 4.8). In some of these embodiments, the MS is in the range of about 0.58 - about 0.68. In presently preferred embodiments, the MS is in the range of 0.58 - 0.68.

[0195] In various embodiments, the MS is at least about 0.55. In certain embodiments, the MS is at least about 0.56, about 0.57, about 0.58, about 0.59, or about 0.60. In certain embodiments, the MS is about 0.70 or less. In specific embodiments, the MS is about 0.69, about 0.68, about 0.67, about 0.66, or about 0.65 or less.

[0196] Further provided herein is a composition produced using one or more of the systems and / or methods provided herein, the composition comprising a mixture of β-cyclodextrin molecules, the mixture of β-cyclodextrin molecules comprising β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). The degree of substitution of the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS.

[0197] In some embodiments, the composition can have an average degree of substitution of from about 7 to about 9, for example, the average degree of substitution can be about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5, about 8.6, about 8.7, about 8.8, about 8.9, or about 9.0. In an exemplary embodiment, the average degree of substitution of the mixture of β-cyclodextrin molecules is about 7.7.

[0198] In some embodiments, the mixture of β-cyclodextrin molecules may contain less than 1% of DS-4. For example, the mixture of β-cyclodextrin molecules may contain about 0.9% of DS-4, about 0.8% of DS-4, about 0.7% of DS-4, about 0.6% of DS-4, about 0.5% of DS-4, about 0.4% of DS-4, about 0.3% of DS-4, about 0.2% of DS-4, or about 0.1% of DS-4. In some aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.9% of DS-4, from about 0.9% to about 0.8% of DS-4, from about 0.8% to about 0.7% of DS-4, from about 0.7% to about 0.6% of DS-4, from about 0.7% to about 0.6% of DS-4, from about 0.6% to about 0.5% of DS-4, from about 0.5% to about 0.4% of DS-4, from about 0.4% to about 0.3% of DS-4, from about 0.3% to about 0.2% of DS-4, from about 0.2% to about 0.1% of DS-4, or less than 0.1% of DS-4. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.8% of DS-4, from less than 1% to about 0.7% of DS-4, from less than 1% to about 0.6% of DS-4, from less than 1% to about 0.5% of DS-4, from less than 1% to about 0.4% of DS-4, from less than 1% to about 0.3% of DS-4, from less than 1% to about 0.2% of DS-4, from less than 1% to about 0.1% of DS-4, from about 0.9% to about 0.1% of DS-4, from about 0.8% to about 0.1% of DS-4, from about 0.7% to about 0.1% of DS-4, from about 0.6% to about 0.1% of DS-4, from about 0.5% to about 0.1% of DS-4, from about 0.4% to about 0.1% of DS-4, or from about 0.3% to about 0.1% of DS-4. In still further aspects, the mixture of β-cyclodextrin may contain less than 1% of DS-4, less than 0.9% of DS-4, less than 0.8% of DS-4, less than 0.7% of DS-4, less than 0.6% of DS-4, less than 0.5% of DS-4, less than 0.4% of DS-4, less than 0.3% of DS-4, less than 0.2% of DS-4, or less than 0.1% of DS-4. In still further aspects, the mixture of β-cyclodextrin molecules may contain about 0.001%, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or about 1% of DS-4.In some embodiments, the amount of DS-4 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-4 in the MALDI-TOF-MS spectrum is 0.73%.

[0199] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 2% to about 5% of DS-5. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 2% to about 2.5% of DS-5, from about 2.5% to about 3% of DS-5, from about 3% to about 3.5% of DS-5, from about 3.5% to about 4% of DS-5, from about 4% to about 4.5% of DS-5, or from about 4.5% to about 5% of DS-5. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 2% to about 3% of DS-5, from about 2% to about 3.5% of DS-5, from about 2% to about 4% of DS-5, from about 2% to about 4.5% of DS-5, from about 2.5% to about 5% of DS-5, from about 3% to about 5% of DS-5, from about 3.5% to about 5% of DS-5, from about 4% to about 5% of DS-5, or from about 3% to about 4% of DS-5. In still further aspects, the mixture of β-cyclodextrin molecules may contain about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9%, about 3.0%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3.7%, about 3.8%, about 3.9%, about 4.0%, about 4.1%, about 4.2%, about 4.3%, about 4.4%, about 4.5%, about 4.6%, about 4.7%, about 4.8%, about 4.9%, or about 5.0% of DS-5. In some embodiments, the amount of DS-5 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-5 in the MALDI-TOF-MS spectrum is 3.49%.

[0200] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 7% to about 13% DS-6. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 7% to about 7.5% DS-6, from about 7.5% to about 8% DS-6, from about 8% to about 8.5% DS-6, from about 8.5% to about 9% DS-6, from about 9% to about 9.5% DS-6, from about 9.5% to about 10% DS-6, from about 10% to about 10.5% DS-6, from about 10.5% to about 11% DS-6, from about 11% to about 11.5% DS-6, from about 11.5% to about 12% DS-6, from about 12% to about 12.5% DS-6, or from about 12.5% to about 13% DS-6. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 7% to about 8% DS-6, from about 7% to about 8.5% DS-6, from about 7% to about 9% DS-6, from about 7% to about 9.5% DS-6, from about 7% to about 10% DS-6, from about 7% to about 10.5% DS-6, from about 7% to about 11% DS-6, from about 7% to about 11.5% DS-6, from about 7% to about 12% DS-6, from about 7% to about 12.5% DS-6, from about 7.5% to about 13% DS-6, from about 8% to about 13% DS-6, from about 8.5% to about 13% DS-6, from about 9% to about 13% DS-6, from about 9.5% to about 13% DS-6, from about 10% to about 13% DS-6, from about 10.5% to about 13% DS-6, from about 11% to about 13% DS-6, from about 11.5% to about 13% DS-6, from about 12% to about 13% DS-6, from about 8% to about 12% DS-6, or from about 9% to about 11% DS-6.In yet a further aspect, the mixture of β-cyclodextrin molecules can contain about 7.0%, about 7.1%, about 7.2%, about 7.3%, about 7.4%, about 7.5%, about 7.6%, about 7.7%, about 7.8%, about 7.9%, about 8.0%, about 8.1%, about 8.2%, about 8.3%, about 8.4%, about 8.5%, about 8.6%, about 8.7%, about 8.8%, about 8.9%, about 9.0%, about 9.1%, about 9.2%, about 9.3%, about 9.4%, about 9.5%, about 9.6%, about 9.7%, about 9.8%, about 9.9%, about 10.0%, about 10.1%, about 10.2%, about 10.3%, about 10.4%, about 10.5%, about 10.6%, about 10.7%, about 10.8%, about 10.9%, about 11.0%, about 11.1%, about 11.2%, about 11.3%, about 11.4%, about 11.5%, about 11.6%, about 11.7%, about 11.8%, about 11.9%, about 12.0%, about 12.1%, about 12.2%, about 12.3%, about 12.4%, about 12.5%, about 12.6%, about 12.7%, about 12.8%, about 12.9%, or about 13.0% of DS-6. In some embodiments, the amount of DS-6 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-6 in the MALDI-TOF-MS spectrum is 10.66%.

[0201] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 21% to about 27% DS-7. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 21% to about 21.5% DS-7, from about 21.5% to about 22% DS-7, from about 22% to about 22.5% DS-7, from about 22.5% to about 23% DS-7, from about 23% to about 23.5% DS-7, from about 23.5% to about 24% DS-7, from about 24% to about 24.5% DS-7, from about 24.5% to about 25% DS-7, from about 25% to about 25.5% DS-7, from about 25.5% to about 26% DS-7, from about 26% to about 26.5% DS-7, or from about 26.5% to about 27% DS-7. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 21% to about 22% DS-7, from about 21% to about 22.5% DS-7, from about 21% to about 23% DS-7, from about 21% to about 23.5% DS-7, from about 21% to about 24% DS-7, from about 21% to about 24.5% DS-7, from about 21% to about 25% DS-7, from about 21% to about 25.5% DS-7, from about 21% to about 26% DS-7, from about 21% to about 26.5% DS-7, from about 21.5% to about 27% DS-7, from about 22% to about 27% DS-7, from 22.5% to about 27% DS-7, from about 23% to about 27% DS-7, from about 23.5% to about 27% DS-7, from about 24% to about 27% DS-7, from about 24.5% to about 27% DS-7, from about 25% to about 27% DS-7, from about 25.5% to about 27% DS-7, from about 26% to about 27% DS-7, from about 22% to about 26% DS-7, or from about 23% to about 25% DS-7.In yet a further aspect, the mixture of β-cyclodextrin molecules can contain about 21.0%, about 21.1%, about 21.2%, about 21.3%, about 21.4%, about 21.5%, about 21.6%, about 21.7%, about 21.8%, about 21.9%, about 22.0%, about 22.1%, about 22.2%, about 22.3%, about 22.4%, about 22.5%, about 22.6%, about 22.7%, about 22.8%, about 22.9%, about 23.0%, about 23.1%, about 23.2%, about 23.3%, about 23.4%, about 23.5%, about 23.6%, about 23.7%, about 23.8%, about 23.9%, about 24.0%, about 24.1%, about 24.2%, about 24.3%, about 24.4%, about 24.5%, about 24.6%, about 24.7%, about 24.8%, about 24.9%, about 25.0%, about 25.1%, about 25.2%, about 25.3%, about 25.4%, about 25.5%, about 25.6%, about 25.7%, about 25.8%, about 25.9%, about 26.0%, about 26.1%, about 26.2%, about 26.3%, about 26.4%, about 26.5%, about 26.6%, about 26.7%, about 26.8%, about 26.9%, or about 27.0% of DS-7. In some embodiments, the amount of DS-7 can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-7 in the MALDI-TOF-MS spectrum is 24.10%.

[0202] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 23% to about 29% DS-8. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 23% to about 23.5% DS-8, from about 23.5% to about 24% DS-8, from about 24% to about 24.5% DS-8, from about 24.5% to about 25% DS-8, from about 25% to about 25.5% DS-8, from about 25.5% to about 26% DS-8, from about 26% to about 26.5% DS-8, from about 26.5% to about 27% DS-8, from about 27% to about 27.5% DS-8, from about 27.5% to about 28% DS-8, from about 28% to about 28.5% DS-8, or from about 28.5% to about 29% DS-8. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 23% to about 24% DS-8, from about 23% to about 24.5% DS-8, from about 23% to about 25% DS-8, from about 23% to about 25.5% DS-8, from about 23% to about 26% DS-8, from about 23% to about 26.5% DS-8, from about 23% to about 27% DS-8, from about 23% to about 27.5% DS-8, from about 23% to about 28% DS-8, from about 23% to about 28.5% DS-8, from about 23.5% to about 29% DS-8, from about 24% to about 29% DS-8, from about 24.5% to about 29% DS-8, from about 25% to about 29% DS-8, from about 25.5% to about 29% DS-8, from about 26% to about 29% DS-8, from about 26.5% to about 29% DS-8, from about 27% to about 29% DS-8, from about 27.5% to about 29% DS-8, from about 28% to about 29% DS-8, from about 24% to about 28% DS-8, or from about 25% to about 27% DS-8.In yet a further aspect, the mixture of β-cyclodextrin molecules can comprise about 23.0%, about 23.1%, about 23.2%, about 23.3%, about 23.4%, about 23.5%, about 23.6%, about 23.7%, about 23.8%, about 23.9%, about 24.0%, about 24.1%, about 24.2%, about 24.3%, about 24.4%, about 24.5%, about 24.6%, about 24.7%, about 24.8%, about 24.9%, about 25.0%, about 25.1%, about 25.2%, about 25.3%, about 25.4%, about 25.5%, about 25.6%, about 25.7%, about 25.8%, about 25.9%, about 26.0%, about 26.1%, about 26.2%, about 26.3%, about 26.4%, about 26.5%, about 26.6%, about 26.7%, about 26.8%, about 26.9%, about 27.0%, about 27.1%, about 27.2%, about 27.3%, about 27.4%, about 27.5%, about 27.6%, about 27.7%, about 27.8%, about 27.9%, about 28.0%, about 28.1%, about 28.2%, about 28.3%, about 28.4%, about 28.5%, about 28.6%, about 28.7%, about 28.8%, about 28.9%, or about 29.0%. In some embodiments, the amount of DS-8 in the composition can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-8 in the MALDI-TOF-MS spectrum is 26.43%.

[0203] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 15% to about 21% DS-9. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 15% to about 15.5% DS-9, from about 15.5% to about 16% DS-9, from about 16% to about 16.5% DS-9, from about 16.5% to about 17% DS-9, from about 17% to about 17.5% DS-9, from about 17.5% to about 18% DS-9, from about 18% to about 18.5% DS-9, from about 18.5% to about 19% DS-9, from about 19% to about 19.5% DS-9, from about 19.5% to about 20% DS-9, from about 20% to about 20.5% DS-9, or from about 20.5% to about 21% DS-9. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 15% to about 16% DS-9, from about 15% to about 16.5% DS-9, from about 15% to about 17% DS-9, from about 15% to about 17.5% DS-9, from about 15% to about 18% DS-9, from about 15% to about 18.5% DS-9, from about 15% to about 19% DS-9, from about 15% to about 19.5% DS-9, from about 15% to about 20% DS-9, from about 15% to about 20.5% DS-9, from about 15.5% to about 21% DS-9, from about 16% to about 21% DS-9, from about 16.5% to about 21% DS-9, from about 17% to about 21% DS-9, from about 17.5% to about 21% DS-9, from about 18% to about 21% DS-9, from about 18.5% to about 21% DS-9, from about 19% to about 21% DS-9, from about 19.5% to about 21% DS-9, from about 20% to about 21% DS-9, from about 16% to about 20% DS-9, or from about 17% to about 19% DS-9.In yet a further aspect, the mixture of β-cyclodextrin molecules can contain about 15.0%, about 15.1%, about 15.2%, about 15.3%, about 15.4%, about 15.5%, about 15.6%, about 15.7%, about 15.8%, about 15.9%, about 16.0%, about 16.1%, about 16.2%, about 16.3%, about 16.4%, about 16.5%, about 16.6%, about 16.7%, about 16.8%, about 16.9%, about 17.0%, about 17.1%, about 17.2%, about 17.3%, about 17.4%, about 17.5%, about 17.6%, about 17.7%, about 17.8%, about 17.9%, about 18.0%, about 18.1%, about 18.2%, about 18.3%, about 18.4%, about 18.5%, about 18.6%, about 18.7%, about 18.8%, about 18.9%, about 19.0%, about 19.1%, about 19.2%, about 19.3%, about 19.4%, about 19.5%, about 19.6%, about 19.7%, about 19.8%, about 19.9%, about 20.0%, about 20.1%, about 20.2%, about 20.3%, about 20.4%, about 20.5%, about 20.6%, about 20.7%, about 20.8%, about 20.9%, or about 21.0% of DS-9. In some embodiments, the amount of DS-9 in the composition can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-9 in the MALDI-TOF-MS spectrum is 18.09%.

[0204] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 6% to about 12% DS-10. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 6% to about 6.5% DS-10, from about 6.5% to about 7% DS-10, from about 7% to about 7.5% DS-10, from about 7.5% to about 8% DS-10, from about 8% to about 8.5% DS-10, from about 8.5% to about 9% DS-10, from about 9% to about 9.5% DS-10, from about 9.5% to about 10% DS-10, from about 10% to about 10.5% DS-10, from about 10.5% to about 11% DS-10, from about 11% to about 11.5% DS-10, or from about 11.5% to about 12% DS-10. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 6% to about 7% DS-10, from about 6% to about 7.5% DS-10, from about 6% to about 8% DS-10, from about 6% to about 8.5% DS-10, from about 6% to about 9% DS-10, from about 6% to about 9.5% DS-10, from about 6% to about 10% DS-10, from about 6% to about 10.5% DS-10, from about 6% to about 11% DS-10, from about 6% to about 11.5% DS-10, from about 6.5% to about 12% DS-10, from about 7% to about 12% DS-10, from about 7.5% to about 12% DS-10, from about 8% to about 12% DS-10, from about 8.5% to about 12% DS-10, from about 9% to about 12% DS-10, from about 9.5% to about 12% DS-10, from about 10% to about 12% DS-10, from about 10.5% to about 12% DS-10, from about 11% to about 12% DS-10, from about 7% to about 11% DS-10, or from about 8% to about 10% DS-10.In yet a further aspect, the mixture of β-cyclodextrin molecules can contain about 6.0%, about 6.1%, about 6.2%, about 6.3%, about 6.4%, about 6.5%, about 6.6%, about 6.7%, about 6.8%, about 6.9%, about 7.0%, about 7.1%, about 7.2%, about 7.3%, about 7.4%, about 7.5%, about 7.6%, about 7.7%, about 7.8%, about 7.9%, about 8.0%, about 8.1%, about 8.2%, about 8.3%, about 8.4%, about 8.5%, about 8.6%, about 8.7%, about 8.8%, about 8.9%, about 9.0%, about 9.1%, about 9.2%, about 9.3%, about 9.4%, about 9.5%, about 9.6%, about 9.7%, about 9.8%, about 9.9%, about 10.0%, about 10.1%, about 10.2%, about 10.3%, about 10.4%, about 10.5%, about 10.6%, about 10.7%, about 10.8%, about 10.9%, about 11.0%, about 11.1%, about 11.2%, about 11.3%, about 11.4%, about 11.5%, about 11.6%, about 11.7%, about 11.8%, about 11.9%, or about 12.0% of DS-10. In some embodiments, the amount of DS-10 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-10 in the MALDI-TOF-MS spectrum is 9.39%.

[0205] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 2% to about 6% DS-11. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 2% to about 2.5% DS-11, from about 2.5% to about 3% DS-11, from about 3% to about 3.5% DS-11, from about 3.5% to about 4% DS-11, from about 4% to about 4.5% DS-11, from about 4.5% to about 5% DS-11, from about 5% to about 5.5% DS-11, or from about 5.5% to about 6% DS-11. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 2% to about 3% DS-11, from about 2% to about 3.5% DS-11, from about 2% to about 4% DS-11, from about 2% to about 4.5% DS-11, from about 2% to about 5% DS-11, from about 2% to about 5.5% DS-11, from about 2.5% to about 6% DS-11, from about 3% to about 6% DS-11, from about 3.5% to about 6% DS-11, from about 4% to about 6% DS-11, from about 4.5% to about 6% DS-11, from about 5% to about 6% DS-11, or from about 3% to about 5% DS-11. In still further aspects, the mixture of β-cyclodextrin molecules may contain about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9%, about 3.0%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3.7%, about 3.8%, about 3.9%, about 4.0%, about 4.1%, about 4.2%, about 4.3%, about 4.4%, about 4.5%, about 4.6%, about 4.7%, about 4.8%, about 4.9%, about 5.0%, about 5.1%, about 5.2%, about 5.3%, about 5.4%, about 5.5%, about 5.6%, about 5.7%, about 5.8%, about 5.9%, or about 6.0% DS-11. In some embodiments, the amount of DS-11 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-11 in the MALDI-TOF-MS spectrum is 4.58%.

[0206] In some embodiments, the mixture of β-cyclodextrin molecules may contain from about 0.5% to about 4% of DS-12. In some aspects, the mixture of β-cyclodextrin molecules may contain from about 0.5% to about 1% of DS-12, from about 1% to about 1.5% of DS-12, from about 1.5% to about 2% of DS-12, from about 2% to about 2.5% of DS-12, from about 2.5% to about 3% of DS-12, from about 3% to about 3.5% of DS-12, or from about 3.5% to about 4% of DS-12. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from about 0.5% to about 1.5% of DS-12, from about 0.5% to about 2% of DS-12, from about 0.5% to about 2.5% of DS-12, from about 0.5% to about 3% of DS-12, from about 0.5% to about 3.5% of DS-12, from about 1% to about 4% of DS-12, from about 1.5% to about 4% of DS-12, from about 2% to about 4% of DS-12, from about 2.5% to about 4% of DS-12, from about 3% to about 4% of DS-12, or from about 1% to about 3% of DS-12. In still further aspects, the mixture of β-cyclodextrin molecules may contain about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9%, about 3.0%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3.7%, about 3.8%, about 3.9%, or about 4.0%. In some embodiments, the amount of DS-12 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In an exemplary embodiment, the area of DS-12 in the MALDI-TOF-MS spectrum is 1.84%.

[0207] In some embodiments, the mixture of β-cyclodextrin molecules may contain less than 1% of DS-13. For example, the mixture of β-cyclodextrin molecules may contain about 0.9% of DS-13, about 0.8% of DS-13, about 0.7% of DS-13, about 0.6% of DS-13, about 0.5% of DS-13, about 0.4% of DS-13, about 0.3% of DS-13, about 0.2% of DS-13, or about 0.1% of DS-13. In some aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.9% of DS-13, about 0.9% to about 0.8% of DS-13, about 0.8% to about 0.7% of DS-13, about 0.7% to about 0.6% of DS-13, about 0.7% to about 0.6% of DS-13, about 0.6% to about 0.5% of DS-13, about 0.5% to about 0.4% of DS-13, about 0.4% to about 0.3% of DS-13, about 0.3% to about 0.2% of DS-13, about 0.2% to about 0.1% of DS-13, or less than 0.1% of DS-13. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.8% of DS-13, from less than 1% to about 0.7% of DS-13, from less than 1% to about 0.6% of DS-13, from less than 1% to about 0.5% of DS-13, from less than 1% to about 0.4% of DS-13, from less than 1% to about 0.3% of DS-13, from less than 1% to about 0.2% of DS-13, from less than 1% to about 0.1% of DS-13, about 0.9% to about 0.1% of DS-13, about 0.8% to about 0.1% of DS-13, about 0.7% to about 0.1% of DS-13, about 0.6% to about 0.1% of DS-13, about 0.5% to about 0.1% of DS-13, about 0.4% to about 0.1% of DS-13, or about 0.3% to about 0.1% of DS-13. In still further aspects, the mixture of β-cyclodextrin may contain less than 1% of DS-13, less than 0.9% of DS-13, less than 0.8% of DS-13, less than 0.7% of DS-13, less than 0.6% of DS-13, less than 0.5% of DS-13, less than 0.4% of DS-13, less than 0.3% of DS-13, less than 0.2% of DS-13, or less than 0.1% of DS-13. In some embodiments, the amount of DS-13 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS.In an exemplary embodiment, the area of DS-13 in the MALDI-TOF-MS spectrum is 0.70%.

[0208] In some embodiments, the composition may contain less than 1% of DS-14. For example, a mixture of β-cyclodextrin molecules may contain about 0.9% of DS-14, about 0.8% of DS-14, about 0.7% of DS-14, about 0.6% of DS-14, about 0.5% of DS-14, about 0.4% of DS-14, about 0.3% of DS-14, about 0.2% of DS-14, or about 0.1% of DS-14. In some aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.9% of DS-14, about 0.9% to about 0.8% of DS-14, about 0.8% to about 0.7% of DS-14, about 0.7% to about 0.6% of DS-14, about 0.7% to about 0.6% of DS-14, about 0.6% to about 0.5% of DS-14, about 0.5% to about 0.4% of DS-14, about 0.4% to about 0.3% of DS-14, about 0.3% to about 0.2% of DS-14, about 0.2% to about 0.1% of DS-14, or less than 0.1% of DS-14. In some additional aspects, the mixture of β-cyclodextrin molecules may contain from less than 1% to about 0.8% of DS-14, from less than 1% to about 0.7% of DS-14, from less than 1% to about 0.6% of DS-14, from less than 1% to about 0.5% of DS-14, from less than 1% to about 0.4% of DS-14, from less than 1% to about 0.3% of DS-14, from less than 1% to about 0.2% of DS-14, from less than 1% to about 0.1% of DS-14, from about 0.9% to about 0.1% of DS-14, from about 0.8% to about 0.1% of DS-14, from about 0.7% to about 0.1% of DS-14, from about 0.6% to about 0.1% of DS-14, from about 0.5% to about 0.1% of DS-14, from about 0.4% to about 0.1% of DS-14, or from about 0.3% to about 0.1% of DS-14. In still further aspects, the mixture of β-cyclodextrin may contain less than 1% of DS-14, less than 0.9% of DS-14, less than 0.8% of DS-14, less than 0.7% of DS-14, less than 0.6% of DS-14, less than 0.5% of DS-14, less than 0.4% of DS-14, less than 0.3% of DS-14, less than 0.2% of DS-14, or less than 0.1% of DS-4 as appropriate. In still further aspects, the mixture of β-cyclodextrin molecules may contain, as appropriate, about 0.001%, about 0.01%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1% of DS-14.In some embodiments, the amount of DS-14 in the mixture of β-cyclodextrin molecules can be determined by MALDI-TOF-MS. In some embodiments, DS-14 is not present in the composition.

[0209] In an exemplary embodiment, the composition comprises a mixture of β-cyclodextrin molecules, the mixture of β-cyclodextrin molecules comprises DS-4, DS-5, DS-6, DS-7, DS-8, DS-9, DS-10, DS-11, DS-12, DS-13, and DS-14, and the mixture of β-cyclodextrin molecules comprises less than 1% of DS-1, DS-2, DS-3, and DS-4.

[0210] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced using one or more of the systems and / or methods provided herein and substituted at one or more hydroxyl positions by hydroxypropyl groups, wherein the mixture comprises less than 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and beta-cyclodextrin having glucose units of the formula:

Chemical formula

[0211] In some embodiments, HP contains one hydroxypropyl group. In some embodiments, HP consists essentially of one hydroxypropyl group. In some embodiments, HP consists of one hydroxypropyl group.

[0212] In some embodiments, R 1 and R 2 Of the total occurrences of the combination of, for example, about 95% or less, such as about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, or about 50% or less are HP.

[0213] R 3 Of the total occurrences of, at least about 5% can be HP. For example, of the total occurrences of R 3 Of the total occurrences of, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, or at least about 10% can be HP.

[0214] In some embodiments, R that is HP 1 and R 2The percentage of the combination is from about 5% to about 95%, for example from about 10% to about 95%, from about 15% to about 95%, from about 20% to about 95%, from about 25% to about 95%, from about 30% to about 95%, from about 35% to about 95%, from about 40% to about 95%, from about 45% to about 95%, from about 50% to about 95%, from about 55% to about 95%, from about 60% to about 95%, from about 65% to about 95%, from about 70% to about 95%, from about 75% to about 95%, from about 80% to about 95%, from about 85% to about 95%, from about 90% to about 95%; for example from about 5% to about 90%, from about 10% to about 90%, from about 15% to about 90%, from about 20% to about 90%, from about 25% to about 90%, from about 30% to about 90%, from about 35% to about 90%, from about 40% to about 90%, from about 45% to about 90%, from about 50% to about 90%, from about 55% to about 90%, from about 60% to about 90%, from about 65% to about 90%, from about 70% to about 90%, from about 75% to about 90%, from about 80% to about 90%, from about 85% to about 90%; for example from about 5% to about 85%, from about 10% to about 85%, from about 15% to about 85%, from about 20% to about 85%, from about 25% to about 85%, from about 30% to about 85%, from about 35% to about 85%, from about 40% to about 85%, from about 45% to about 85%, from about 50% to about 85%, from about 55% to about 85%, from about 60% to about 85%, from about 65% to about 85%, from about 70% to about 85%, from about 75% to about 85%, from about 80% to about 85%; for example from about 5% to about 80%, from about 10% to about 80%, from about 15% to about 80%, from about 20% to about 80%, from about 25% to about 80%, from about 30% to about 80%, from about 35% to about 80%, from about 40% to about 80%, from about 45% to about 80%, from about 50% to about 80%, from about 55% to about 80%, from about 60% to about 80%, from about 65% to about 80%, from about 70% to about 80%, from about 75% to about 80%; for example from about 5% to about 75%, from about 10% to about 75%, from about 15% to about 75%, from about 20% to about 75%, from about 25% to about 75%, from about 30% to about 75%, from about 35% to about 75%, from about 40% to about 75%, from about 45% to about 75%, from about 50% to about 75%, from about 55% to about 75%, from about 60% to about 75%, from about 65% to about 75%, from about 70% to about 75%; for example from about 5% to about 70%, from about 10% to about 70%, from about 15% to about 70%, from about 20% to about 70%, from about 25% to about 70%, from about 30% to about 70%, from about 35% to about 70%, from about 40% to about 70%, from about 45% to about 70%, from about 50% to about 70%, from about 55% to about 70%, from about 60% to about 70%, from about 65% to about 70%;For example, from about 5% to about 65%, from about 10% to about 65%, from about 15% to about 65%, from about 20% to about 65%, from about 25% to about 65%, from about 30% to about 65%, from about 35% to about 65%, from about 40% to about 65%, from about 45% to about 65%, from about 50% to about 65%, from about 55% to about 65%, from about 60% to about 65%; for example, from about 5% to about 60%, from about 10% to about 60%, from about 15% to about 60%, from about 20% to about 60%, from about 25% to about 60%, from about 30% to about 60%, from about 35% to about 60%, from about 40% to about 60%, from about 45% to about 60%, from about 50% to about 60%, from about 55% to about 60%; for example, from about 5% to about 55%, from about 10% to about 55%, from about 15% to about 55%, from about 20% to about 55%, from about 25% to about 55%, from about 30% to about 55%, from about 35% to about 55%, from about 40% to about 55%, from about 45% to about 55%, from about 50% to about 55%; for example, from about 5% to about 50%, from about 10% to about 50%, from about 15% to about 50%, from about 20% to about 50%, from about 25% to about 50%, from about 30% to about 50%, from about 35% to about 50%, from about 40% to about 50%, from about 45% to about 50%; for example, from about 5% to about 45%, from about 10% to about 45%, from about 15% to about 45%, from about 20% to about 45%, from about 25% to about 45%, from about 30% to about 45%, from about 35% to about 45%, from about 40% to about 45%; for example, from about 5% to about 40%, from about 10% to about 40%, from about 15% to about 40%, from about 20% to about 40%, from about 25% to about 40%, from about 30% to about 40%, from about 35% to about 40%; for example, from about 5% to about 35%, from about 10% to about 35%, from about 15% to about 35%, from about 20% to about 35%, from about 25% to about 35%, from about 30% to about 35%; for example, from about 5% to about 30%, from about 10% to about 30%, from about 15% to about 30%, from about 20% to about 30%, from about 25% to about 30%; for example, from about 5% to about 25%, from about 10% to about 25%, from about 15% to about 25%, from about 20% to about 25%; for example, from about 5% to about 20%, from about 10% to about 20%, from about 15% to about 20%; for example, from about 5% to about 15%, from about 10% to about 15%; or ranges from about 5% to about 10%.;

[0215] The mixture may contain less than 0.1% of DS-0 and less than 0.1% of DS-1 combined. For example, the mixture may contain less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-0, and / or the mixture may contain less than 0.1%, less than 0.09%, less than 0.08%, less than 0.07%, less than 0.06%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, or less than 0.01% of DS-1.

[0216] The amount of DS-0 or DS-1 can be determined by the peak height of the electrospray MS spectrum.

[0217] The mixture may have an average molar substitution in the range of about 0.40 to about 0.80. For example, the mixture may have an average molar substitution of about 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, or about 0.80. The mixture may have an average degree of substitution (「DS a 」) in the range of about 3 to about 7, about 4 to about 7, about 5 to about 7, or about 6 to about 7. For example, the mixture may have an average degree of substitution of about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, or about 7. In other words, the average number of occurrences of HP per beta-cyclodextrin can be 3 to 4, 3 to 5, 3 to 6, 3 to 7, 4 to 5, 4 to 6, 4 to 7, 5 to 6, 5 to 7, or 6 to 7.

[0218] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC or gas chromatography.

[0219] The composition may contain 0.01% or less of propylene glycol. For example, the composition may contain 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, or about 0.001% or less of propylene glycol. The amount of propylene glycol can be measured by HPLC, gas chromatography, or the PG / EG ratio of propylene glycol to ethylene glycol.

[0220] The composition may contain 1 ppm or less of propylene oxide, 0.9 ppm or less of propylene oxide, 0.8 ppm or less of propylene oxide, 0.7 ppm or less of propylene oxide, 0.6 ppm or less of propylene oxide, 0.5 ppm or less of propylene oxide, 0.4 ppm or less of propylene oxide, 0.3 ppm or less of propylene oxide, 0.2 ppm or less of propylene oxide, or 0.1 ppm or less of propylene oxide. The amount of propylene oxide can be measured by HPLC or gas chromatography.

[0221] The total amount of other unspecified impurities in the composition can be 0.05% or less. For example, the total amount of unspecified impurities in the composition can be 0.05%, less than 0.05%, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less. The amount of unspecified impurities can be measured by HPLC or gas chromatography.

[0222] The composition may be suitable for intrathecal, intravenous, or intraventricular administration to a patient in need thereof. The patient can be an adult patient or a pediatric patient. The composition may further contain a pharmaceutically acceptable diluent.

[0223] The composition can solubilize lipids in an aqueous medium. The lipids can include non-esterified or esterified cholesterol. The composition may be provided as a solution having a solution concentration of 20 w / v% of a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions. The composition can have an affinity for non-esterified cholesterol. The solubilization can be determined by UV spectroscopy or HPLC.

[0224] In some embodiments, about 200 mg of the composition solubilizes at least about 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or at least about 10 mg of non-esterified cholesterol in distilled water at room temperature. In one example, 1 mL of the solution can solubilize about 2 mg of non-esterified cholesterol at room temperature as measured by UV spectroscopy after about 24 hours.

[0225] The composition can have a concentration in solution of from about 10 mg / mL to about 200 mg / mL. For example, the composition can have a concentration in solution of from about 10 mg / mL to about 20 mg / mL, from about 10 mg / mL to about 30 mg / mL, from about 10 mg / mL to about 40 mg / mL, from about 10 mg / mL to about 50 mg / mL, from about 10 mg / mL to about 60 mg / mL, from about 10 mg / mL to about 70 mg / mL, from about 10 mg / mL to about 80 mg / mL, from about 10 mg / mL to about 90 mg / mL, from about 10 mg / mL to about 100 mg / mL, from about 10 mg / mL to about 110 mg / mL, from about 10 mg / mL to about 120 mg / mL, from about 10 mg / mL to about 130 mg / mL, from about 10 mg / mL to about 140 mg / mL, from about 10 mg / mL to about 150 mg / mL, from about 10 mg / mL to about 160 mg / mL, from about 10 mg / mL to about 170 mg / mL, from about 10 mg / mL to about 180 mg / mL, from about 10 mg / mL to about 190 mg / mL, from about 20 mg / mL to about 200 mg / mL, from about 30 mg / mL to about 200 mg / mL, from about 40 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 110 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 130 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 150 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 170 mg / mL to about 200 mg / mL, from about 180 mg / mL to about 200 mg / mL, or from about 190 mg / mL to about 200 mg / mL.

[0226] Further provided herein is a composition comprising a mixture of beta-cyclodextrin molecules produced by any of the systems and / or processes described and provided herein and substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises less than 0.05% unsubstituted beta-cyclodextrin ("DS-0") and less than 0.05% beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1") and has an average degree of substitution of 6.02 to 7.98, and the composition is suitable for intrathecal, intravenous, oral, or intraventricular administration to a patient in need thereof. In some embodiments, the composition has a pH of 6.0 to 7.9. In some embodiments, the true density of the composition is about 1.096 to 1.098 g / cm 3It is so. In some embodiments, the weight osmolality of the composition is about 635 - 695 mOs / kg. In some embodiments, the composition further comprises a container and invisible particulate matter, and the amount of invisible particulate matter with a size of 25 microns or more is 600 / container or less. In some embodiments, the composition contains propylene glycol at 10 ppb or less when measured by HPLC. In some embodiments, the composition contains propylene glycol at 10 ppb or less when measured by gas chromatography. In some embodiments, the composition contains propylene glycol at 10 ppb or less when measured by the PG / EG ratio of propylene glycol to ethylene glycol. In some embodiments, the composition contains propylene oxide at 1 ppm or less. In some embodiments, the total amount of other unspecified impurities is 0.05% or less as measured by HPLC. In some embodiments, the composition has a concentration of about 10 mg / mL to about 200 mg / mL. In some embodiments, the composition exhibits lower toxicity than Trappsol® Cyclo. In some embodiments, the composition has a conductivity of about 200 μS / cm or less. In some embodiments, the composition is stable for at least 6 months. In some embodiments, the composition further comprises at least one of a pharmaceutical excipient, a carrier, a pharmaceutically acceptable diluent, a pH adjuster, and a buffer. In some aspects, the pH adjuster is sodium hydroxide. In some aspects, the buffer comprises sodium dihydrogen phosphate and disodium hydrogen phosphate.

[0227] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of β-cyclodextrin molecules, the mixture of β-cyclodextrin molecules comprising β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”), the mixture of β-cyclodextrin molecules comprising less than 1% DS-4. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 0.5 w / w% to about 1 w / w% DS-4. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 2 w / w% to about 5 w / w% DS-5. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 7 w / w% to about 13 w / w% DS-6. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 21 w / w% to about 27 w / w% DS-7. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 23 w / w% to about 29 w / w% DS-8. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 15 w / w% to about 21 w / w% DS-9. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 6 w / w% to about 12 w / w% DS-10.In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 2 w / w% to about 6 w / w% of DS-11. In some embodiments, the mixture of β-cyclodextrin molecules comprises from about 0.5 w / w% to about 4 w / w% of DS-12. In some embodiments, the mixture of β-cyclodextrin molecules comprises less than about 1 w / w% of DS-13. In some embodiments, the mixture of β-cyclodextrin molecules is suitable for intravenous, intrathecal, or intraventricular administration. In some embodiments, the amounts of DS-1, DS-2, DS-3, DS-4, DS-5, DS-6, DS-7, DS-8, DS-9, DS-10, DS-11, DS-12, and DS-13 in the mixture of β-cyclodextrin molecules are determined by MALDI-TOF-MS. In some embodiments, DS-8 has the highest concentration in the mixture of β-cyclodextrin molecules when compared to the concentrations of DS-1, DS-2, DS-3, DS-4, DS-5, DS-6, DS-7, DS-9, DS-10, DS-11, DS-12, and DS-13. In some embodiments, the β-cyclodextrin molecules are substituted at the 2-O-position at a rate of 35 to 55%, at the 3-O-position at a rate of 45 to 65%, and at the 6-O-position at a rate of 0 to 20%. In some embodiments, the substitution rates at the 2-O-position, 3-O-position, and 6-O-position are determined by DEPT-edited HSQC. In some embodiments, the composition has an average degree of substitution of from about 7 to about 9. In an exemplary embodiment, the composition has an average degree of substitution of about 7.7. In some embodiments, the composition has a density of about 1.095 g / cm³. 3 to about 1.100 g / cm³ 3It has a true density. In some embodiments, the composition has a weight osmolality of from about 600 mOs / kg to about 750 mOs / kg. In some embodiments, the composition is a clear and colorless solution. In some embodiments, the composition has a pH of from about 4.0 to about 6.0. In some embodiments, the composition has a viscosity of from 1.5 cP to about 3.0 cP at 20°C. In some embodiments, the composition contains less than about 0.05% impurities. In some embodiments, the composition contains less than 600 particles per container having a diameter of 25 microns or more. In some embodiments, the composition contains less than 6000 particles per container having a diameter of 10 microns or more.

[0228] Further provided herein is a composition produced by any of the systems and / or methods provided herein and containing a mixture of β-cyclodextrin molecules, the composition having at least one peak at about 5.0 to 5.4 ppm corresponding to the anomeric proton of the β-cyclodextrin molecule, at least one peak at about 3.2 to 4.2 ppm corresponding to the proton within the core region of the β-cyclodextrin molecule, and at least one peak at about 1.0 to 1.2 ppm corresponding to the methyl proton of the side chain of the β-cyclodextrin molecule 1 having an 1H-NMR spectrum.

[0229] Further provided herein is a composition produced by any one of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules containing less than 1% of β-cyclodextrin substituted with four hydroxypropyl groups (“DS-4”). In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on the area percentage from the MALDI-TOF-MS spectrum. In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on weight percentage. In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with three hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with two hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with one hydroxypropyl group (“DS-1”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 1% to about 5% of β-cyclodextrin substituted with five hydroxypropyl groups (“DS-5”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises from about 7% to about 13% of β-cyclodextrin substituted with six hydroxypropyl groups (“DS-6”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises from about 8% to about 12% of DS-6. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises from about 16% to about 22% of β-cyclodextrin substituted with seven hydroxypropyl groups (“DS-7”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises from about 17% to about 21% of DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises from about 26% to about 32% of β-cyclodextrin substituted with eight hydroxypropyl groups (“DS-8”).In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains about 27% to about 31% DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains about 22% to about 28% of β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains about 23% to about 27% DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains about 11% to about 17% of β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains about 12% to about 16% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains less than 1% of β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains less than 1% of β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 6.4 to about 7.0. In an exemplary embodiment, the average degree of substitution is about 6.69. In some embodiments, about 52% to about 58% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, about 55% to about 56% of the hydroxypropyl substitution in the β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, about 41% to about 47% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position.In some embodiments, about 43% to about 45% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the concentration of the composition does not substantially change the time required for nanofiltration. In some embodiments, the length of time for nanofiltration of the composition ranges from 1.04 to 1.20 hours per diafiltration volume (kg solution / m2-hr / L solution). In some embodiments, the composition has a conductivity of 0 to 8.0 μS / cm, 0 to 4.5 μS / cm, 0 to 3 μS / cm, or 0 to 1.5 μS / cm.

[0230] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprising β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), and β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), the composition comprising less than 1% of β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”) and less than 1% of β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”). In some embodiments, the composition comprises 0.0-1.0% of β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), 0.0-1.0% of β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and 0.0-1.0% of β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, DS-8 has the highest concentration in the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules when compared to DS-5, DS-6, DS-7, DS-9, and DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises about 1%-about 5% of DS-5. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin comprises about 7%-about 13% of DS-6.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains from about 16% to about 22% DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains from about 26% to about 32% DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains from about 22% to about 28% DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin contains from about 11% to about 17% DS-10. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 6.4 to about 7.0. In an exemplary embodiment, the average degree of substitution is about 6.69. In some embodiments, from about 52% to about 58% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 41% to about 47% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the composition has an -ESI-MS spectrum with peaks at about 653 m / z, about 682 m / z, about 711 m / z, about 741 m / z, about 769 m / z, about 799 m / z, about 828 m / z, and about 857 m / z, and a +ESI-MS spectrum with peaks at about 686 m / z, about 715 m / z, about 744 m / z, about 773 m / z, about 802 m / z, about 832 m / z, about 861 m / z, and about 890 m / z. In some embodiments, the composition has a MALDI-TOF spectrum with peaks at about 1436 m / z, about 1495 m / z, about 1555 m / z, about 1614 m / z, about 1674 m / z, and about 1733 m / z. In some embodiments, the weight osmolality of the composition is from about 635 to 695 mOs / kg. In some embodiments, the true density of the composition is from about 1.096 to 1.098 g / cm. 3It is so. In some embodiments, the composition contains propylene glycol of 10 ppb or less when measured by HPLC. In some embodiments, the composition contains propylene oxide of 1 ppm or less. In some embodiments, the total amount of other unspecified impurities is 0.05% or less as measured by HPLC. In some embodiments, the composition further contains 0 to 10 ppm of chloride. In some embodiments, the composition is nanofiltrated. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in HPLC-ELSD after nanofiltration. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in NMR after nanofiltration.

[0231] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules containing less than 1% of hydroxypropyl-β-cyclodextrin having 5 hydroxypropyl groups (“DS-5”). In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on the area percentage from the MALDI-TOF-MS spectrum. In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on weight percentage. In some embodiments, the composition comprises β-cyclodextrin substituted with less than 1% of 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 0% to about 6% of hydroxypropyl-β-cyclodextrin having 6 hydroxypropyl groups (“DS-6”). In some aspects, the mixture of isomerically purified β-hydroxypropyl cyclodextrin molecules comprises from about 1% to about 5% of DS-6. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 8% to about 14% of hydroxypropyl-β-cyclodextrin having 7 hydroxypropyl groups (“DS-7”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 9% to about 13% of DS-7. In some embodiments, the mixture of isomerically purified β-hydroxypropyl cyclodextrin molecules comprises from about 19% to about 25% of hydroxypropyl-β-cyclodextrin having 8 hydroxypropyl groups (“DS-8”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 20% to about 24% of DS-8.In some embodiments, the mixture of isomerically purified β-hydroxypropyl cyclodextrin molecules comprises hydroxypropyl β-cyclodextrin substituted with about 23% to about 29% nine hydroxypropyl groups (“DS-9”). In some aspects, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises about 24% to about 28% DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises hydroxypropyl β-cyclodextrin substituted with about 17% to about 23% ten hydroxypropyl groups (“DS-10”). In some aspects, the mixture of isomerically purified β-hydroxypropyl cyclodextrin molecules comprises about 18% to about 22% DS-10. In some embodiments, the mixture of isomerically purified β-hydroxypropyl cyclodextrin molecules comprises hydroxypropyl β-cyclodextrin substituted with about 9% to about 15% eleven hydroxypropyl groups (“DS-11”). In some aspects, the mixture of isomerically purified β-cyclodextrin molecules comprises about 10% to about 14% DS-11. In some embodiments, the mixture of isomerically purified β-cyclodextrin molecules comprises hydroxypropyl β-cyclodextrin substituted with about 2% to about 8% twelve hydroxypropyl groups (“DS-12”). In some aspects, the mixture of isomerically purified β-cyclodextrin molecules comprises about 3% to about 7% DS-12. In some embodiments, the mixture of isomerically purified β-cyclodextrin molecules has an average degree of substitution of about 7 to about 8. In an exemplary embodiment, the average degree of substitution is about 7.42. In some embodiments, about 36% to about 42% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecules is located at the 3-O-position. In some aspects, about 37% to about 41% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecules is located at the 3-O-position. In some embodiments, about 58% to about 64% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecules is located at the 2-O-position.In some embodiments, about 59% to about 63% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the concentration of the composition does not substantially change the time required for nanofiltration. In some embodiments, the length of time for nanofiltration of the composition ranges from 1.04 to 1.20 hours per diafiltration volume (kg solution / m2-hr / L solution). In some embodiments, no substantial difference is observed in HPLC-ELSD after nanofiltration of the composition compared to before nanofiltration. In some embodiments, no substantial difference is observed in NMR after nanofiltration of the composition compared to before nanofiltration. In some embodiments, the composition has a conductivity of 0 to 8.0 μS / cm, 0 to 4.5 μS / cm, 0 to 3 μS / cm, or 0 to 1.5 μS / cm.

[0232] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprising β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), and β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), the composition comprising less than 1% of β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), and the composition comprising less than 1% of β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”) and hydroxypropyl-β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, DS-9 has the highest concentration in the composition when compared to DS-6, DS-7, DS-8, DS-10, DS-11, and DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 0% to about 6% of DS-6.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 8% to about 14% DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 19% to about 25% DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 23% to about 29% DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 17% to about 23% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 9% to about 15% DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 2% to about 8% DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules has an average degree of substitution of from about 7 to about 8. In some embodiments, from about 36% to about 42% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 58% to about 64% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the composition has an -ESI-MS spectrum with peaks at about 682 m / z, about 712 m / z, about 740 m / z, about 770 m / z, about 798 m / z, about 828 m / z, about 856 m / z, and about 886 m / z, and a +ESI-MS spectrum with peaks at about 744 m / z, about 773 m / z, about 803 m / z, about 832 m / z, about 860 m / z, about 889 m / z, and about 919 m / z. In some embodiments, the composition has a MALDI-TOF-MS spectrum with peaks at about 1497 m / z, about 1557 m / z, about 1616 m / z, about 1675 m / z, about 1734 m / z, about 1794 m / z, and about 1914 m / z. In some embodiments, the weight osmolality of the composition is from about 635 to 695 mOs / kg. In some embodiments, the true density of the composition is from about 1.096 to 1.098 g / cm. 3It is so. In some embodiments, the composition contains propylene glycol of 10 ppb or less when measured by HPLC. In some embodiments, the composition contains propylene oxide of 1 ppm or less. In some embodiments, the total amount of other unspecified impurities is 0.05% or less as measured by HPLC. In some embodiments, the composition contains chlorides of 0 to 10 ppm. In some embodiments, the composition has a conductivity of 0 to 8 μS / cm. In some embodiments, the composition is nanofiltrated. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in HPLC-ELSD after nanofiltration. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in NMR after nanofiltration.

[0233] Further provided herein is a composition produced by any of the methods and / or systems provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprising less than 1% of hydroxypropyl-β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”) and less than 1% of β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on the area percentage from a MALDI-TOF-MS spectrum. In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on weight percentage. In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 1% to about 7% of β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 2% to about 6% of DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 16% to about 22% of β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 17% to about 21% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 22% to about 28% of β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”).In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 23% to about 27% DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 19% to about 25% β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 20% to about 24% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 14% to about 20% β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 15% to about 19% DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 5% to about 11% β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 6% to about 10% DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 1% to about 7% β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”). In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 2% to about 6% DS-13. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 8 to about 9. In an exemplary embodiment, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 8.53. In some embodiments, from about 26% to about 32% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 27% to about 31% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position.In some embodiments, about 68% to about 74% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some aspects, about 69% to about 73% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the concentration of the composition does not substantially change the time required for nanofiltration. In some aspects, the length of time for nanofiltration of the composition is in the range of 1.04 to 1.20 hours per diafiltration volume (kg solution / m. 2 -hr / L solution). In some embodiments, no substantial difference is observed in HPLC-ELSD after nanofiltration of the composition compared to before nanofiltration. In some embodiments, no substantial difference is observed in NMR after nanofiltration of the composition compared to before nanofiltration. In some embodiments, the composition has a conductivity of 0 to 8.0 μS / cm, 0 to 4.5 μS / cm, 0 to 3 μS / cm, or 0 to 1.5 μS / cm.

[0234] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprising β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), and β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), the composition comprising less than 1% of β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”) and less than 1% of β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, DS-9 has the highest concentration in the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules when compared to DS-6, DS-7, DS-8, DS-10, DS-11, DS-12, and DS-13. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 16% to about 22% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 22% to about 28% of DS-9.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 19% to about 25% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 14% to about 20% DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 5% to about 11% DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 1% to about 7% DS-13. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 8 to about 9. In an exemplary embodiment, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 8.53. In some embodiments, from about 26% to about 32% of the hydroxypropyl substitution in the β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 68% to about 74% of the hydroxypropyl substitution in the β-cyclodextrin molecule is located at the 2-O-position. In an exemplary embodiment, the HPLC-CAD average retention time of the composition is about 13.5 minutes. In some embodiments, the composition has an -ESI-MS spectrum with peaks at about 741 m / z, about 769 m / z, about 799 m / z, about 828 m / z, about 856 m / z, about 886 m / z, and a +ESI-MS spectrum with peaks at about 773 m / z, about 803 m / z, about 833 m / z, about 860 m / z, about 889 m / z, and about 920 m / z. In some embodiments, the composition has a MALDI-TOF spectrum with peaks at about 1557 m / z, about 1617 m / z, about 1676 m / z, about 1736 m / z, about 1795 m / z, about 1855 m / z, and about 1915 m / z. In some embodiments, the weight osmolality of the composition is from about 635 to 695 mOs / kg. In some embodiments, the true density of the composition is from about 1.096 to 1.098 g / cm. 3It is. In some embodiments, the composition contains propylene glycol of 10 ppb or less when measured by HPLC. In some embodiments, the composition contains propylene oxide of 1 ppm or less. In some embodiments, the total amount of other unspecified impurities is 0.05% or less as measured by HPLC. In some embodiments, the composition contains chlorides of 0 to 10 ppm. In some embodiments, the composition contains chlorides of 0 to 1 ppm. In some embodiments, the composition has a conductivity of 0 to 8 μS / cm. In some embodiments, the composition is nanofiltrated. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in HPLC-ELSD after nanofiltration. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in NMR after nanofiltration.

[0235] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules containing less than 1% of hydroxypropyl-β-cyclodextrin having six hydroxypropyl groups (“DS-6”). In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on the area percentage from a MALDI-TOF-MS spectrum. In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on weight percentage. In some embodiments, the composition comprises β-cyclodextrin substituted with less than 1% of five hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with four hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with three hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with two hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with one hydroxypropyl group (“DS-1”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 0% to about 6% of β-cyclodextrin substituted with seven hydroxypropyl groups (“DS-7”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 1% to about 5% of DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 13% to about 19% of β-cyclodextrin substituted with eight hydroxypropyl groups (“DS-8”). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 14% to about 18% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 22% to about 28% of β-cyclodextrin substituted with nine hydroxypropyl groups (“DS-9”). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 23% to about 27% of DS-9.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 23% to about 29% β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”). In some aspects, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 24% to about 28% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 12% to about 18% β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”). In some aspects, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 13% to about 17% DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 7% to about 13% β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”). In some aspects, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 8% to about 12% DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 2% to about 8% β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”). In some aspects, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 3% to about 7% DS-13. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 7.5 to about 8.5. In an exemplary embodiment, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 8.08. In some embodiments, from about 22% to about 28% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some aspects, from about 23% to about 27% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position.In some embodiments, about 72% to about 78% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some aspects, about 73% to about 77% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the concentration of the composition does not substantially change the time required for nanofiltration. In some aspects, the length of time for nanofiltration of the composition is in the range of 1.04 to 1.20 hours per diafiltration volume (kg solution / m. 2 -hr / L solution). In some embodiments, no substantial difference is observed in HPLC-ELSD after nanofiltration of the nanofiltrated composition as compared to before nanofiltration. In some embodiments, no substantial difference is observed in NMR after nanofiltration of the nanofiltrated composition as compared to before nanofiltration. In some embodiments, the composition has a conductivity of 0 to 8.0 μS / cm, 0 to 4.5 μS / cm, 0 to 3 μS / cm, or 0 to 1.5 μS / cm.

[0236] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules, including β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”), β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”), the composition comprising less than 1% of β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, DS-9 has the highest concentration in the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules when compared to DS-7, DS-8, DS-10, DS-11, DS-12, DS-13, and DS-14. In some embodiments, DS-10 has the highest concentration in the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules when compared to DS-7, DS-8, DS-10, DS-11, DS-12, DS-13, and DS-14.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 0% to about 6% of DS-7. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 13% to about 19% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 22% to about 28% of DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 23% to about 29% of DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 12% to about 18% of DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 7% to about 13% of DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 2% to about 8% of DS-13. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 0% to about 6% of DS-14. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 7.5 to about 8.5. In some embodiments, from about 22% to about 28% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 72% to about 78% of the hydroxypropyl substitution in the β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the composition has an -ESI-MS spectrum with peaks at about 740 m / z, about 770 m / z, about 798 m / z, about 828 m / z, and about 857 m / z, and a +ESI-MS spectrum with peaks at about 803 m / z, about 831 m / z, about 861 m / z, about 889 m / z, and about 919 m / z. In some embodiments, the composition has a MALDI-TOF spectrum with peaks at about 1559 m / z, about 1618 m / z, about 1678 m / z, about 1737 m / z, about 1796 m / z, about 1857 m / z, and about 1916 m / z.In some embodiments, the weight osmolality of the composition is about 635 - 695 mOs / kg. In some embodiments, the true density of the composition is about 1.096 - 1.098 g / cm³. 3 In some embodiments, the composition contains less than 10 ppb of propylene glycol as measured by HPLC. In some embodiments, the composition contains less than 1 ppm of propylene oxide. In some embodiments, the total amount of other unspecified impurities is less than 0.05% as measured by HPLC. In some embodiments, the composition contains 0 - 10 ppm of chloride. In some embodiments, the composition has a conductivity of 0 - 8 μS / cm. In some embodiments, the composition is nanofiltrated. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in HPLC-ELSD after nanofiltration. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in NMR after nanofiltration.

[0237] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules containing less than 1% of hydroxypropyl-β-cyclodextrin having 7 hydroxypropyl groups ( "DS-7"). In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on the area percentage from the MALDI-TOF-MS spectrum. In some embodiments, the percentage of hydroxypropyl-β-cyclodextrin is based on weight percentage. In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 6 hydroxypropyl groups ( "DS-6"), β-cyclodextrin substituted with 5 hydroxypropyl groups ( "DS-5"), β-cyclodextrin substituted with 4 hydroxypropyl groups ( "DS-4"), β-cyclodextrin substituted with 3 hydroxypropyl groups ( "DS-3"), β-cyclodextrin substituted with 2 hydroxypropyl groups ( "DS-2"), and β-cyclodextrin substituted with 1 hydroxypropyl group ( "DS-1"). In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 6% to about 12% of β-cyclodextrin substituted with 8 hydroxypropyl groups ( "DS-8"). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 7% to about 11% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 18% to about 24% of β-cyclodextrin substituted with 9 hydroxypropyl groups ( "DS-9"). In some aspects, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 19% to about 23% of DS-9. In some embodiments, the mixture of isomerically purified hydroxypropyl-β-cyclodextrin molecules comprises from about 24% to about 30% of β-cyclodextrin substituted with 10 hydroxypropyl groups ( "DS-10").In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 25% to about 29% DS-10. In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 18% to about 24% β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”). In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 19% to about 23% DS-11. In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 10% to about 16% β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”). In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 11% to about 15% DS-12. In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 4% to about 10% β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”). In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 5% to about 9% DS-13. In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 0% to about 6% β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”). In some embodiments, a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains from about 1% to about 5% DS-14. In some embodiments, the average degree of substitution of a mixture of isomerically purified hydroxypropyl β-cyclodextrin is from about 9 to about 10. In an exemplary embodiment, the average degree of substitution of a mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 9.65. In some embodiments, from about 15% to about 21% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, from about 16% to about 20% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position.In some embodiments, about 79% to about 85% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some aspects, about 80% to about 84% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the concentration of the composition does not substantially change the time required for nanofiltration. In some embodiments, the length of time for nanofiltration of the composition is in the range of 1.04 to 1.20 hours per diafiltration volume (kg solution / m. 2 -hr / L solution). In some embodiments, no substantial difference is observed in HPLC-ELSD after nanofiltration of the composition compared to before nanofiltration. In some embodiments, no substantial difference is observed in NMR after nanofiltration of the composition compared to before nanofiltration. In some embodiments, the composition has a conductivity of 0 to 8.0 μS / cm, 0 to 4.5 μS / cm, 0 to 3 μS / cm, or 0 to 1.5 μS / cm.

[0238] Further provided herein is a composition produced by any of the systems and / or methods provided herein, the composition comprising a mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprising β-cyclodextrin substituted with 8 hydroxypropyl groups (“DS-8”), β-cyclodextrin substituted with 9 hydroxypropyl groups (“DS-9”), β-cyclodextrin substituted with 10 hydroxypropyl groups (“DS-10”), β-cyclodextrin substituted with 11 hydroxypropyl groups (“DS-11”), β-cyclodextrin substituted with 12 hydroxypropyl groups (“DS-12”), β-cyclodextrin substituted with 13 hydroxypropyl groups (“DS-13”), and β-cyclodextrin substituted with 14 hydroxypropyl groups (“DS-14”), the composition comprising less than 1% of β-cyclodextrin substituted with 7 hydroxypropyl groups (“DS-7”). In some embodiments, the composition comprises less than 1% of β-cyclodextrin substituted with 6 hydroxypropyl groups (“DS-6”), 1% of β-cyclodextrin substituted with 5 hydroxypropyl groups (“DS-5”), β-cyclodextrin substituted with 4 hydroxypropyl groups (“DS-4”), β-cyclodextrin substituted with 3 hydroxypropyl groups (“DS-3”), β-cyclodextrin substituted with 2 hydroxypropyl groups (“DS-2”), and β-cyclodextrin substituted with 1 hydroxypropyl group (“DS-1”). In some embodiments, DS-10 has the highest concentration in the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules when compared to DS-8, DS-9, DS-11, DS-12, DS-13, and DS-14. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 6% to about 12% of DS-8. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules comprises from about 18% to about 24% of DS-9.In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains about 24% to about 30% DS-10. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains about 18% to about 24% DS-11. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains about 10% to about 16% DS-12. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains about 4% to about 10% DS-13. In some embodiments, the mixture of isomerically purified hydroxypropyl β-cyclodextrin molecules contains about 0% to about 6% DS-14. In some embodiments, the average degree of substitution of the mixture of isomerically purified hydroxypropyl β-cyclodextrin is about 9 to about 10. In some embodiments, about 15% to about 21% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 3-O-position. In some embodiments, about 79% to about 85% of the hydroxypropyl substitution in the hydroxypropyl β-cyclodextrin molecule is located at the 2-O-position. In some embodiments, the composition has an -ESI-MS spectrum with peaks at about 770 m / z, about 798 m / z, about 828 m / z, about 857 m / z, about 885 m / z, and a +ESI-MS spectrum with peaks at about 803 m / z, about 831 m / z, about 861 m / z, about 889 m / z, and about 919 m / z. In some embodiments, the composition has a MALDI-TOF spectrum with peaks at about 1614 m / z, about 1673 m / z, about 1733 m / z, about 1792 m / z, about 1852 m / z, about 1912 m / z, and about 1971 m / z. In some embodiments, the weight osmolality of the composition is about 635 - 695 mOs / kg. In some embodiments, the true density of the composition is about 1.096 - 1.098 g / cm. 3It is. In some embodiments, the composition contains propylene glycol of 10 ppb or less when measured by HPLC. In some embodiments, the composition contains propylene oxide of 1 ppm or less. In some embodiments, the total amount of other unspecified impurities is 0.05% or less as measured by HPLC. In some embodiments, the composition contains chlorides of 0 to 10 ppm. In some embodiments, the composition has a conductivity of 0 to 8 μS / cm. In some embodiments, the composition is nanofiltrated. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in HPLC-ELSD after nanofiltration. In some embodiments, for the nanofiltrated composition, when compared with that before nanofiltration, no substantial difference is observed in NMR after nanofiltration.

[0239] The systems and methods provided herein can be used, for example, to produce the compositions described in U.S. Patent No. 10,933,083, filed on March 2, 2021, and its related applications (e.g., U.S. Patent No. 9,675,634, filed on June 13, 2017; U.S. Patent No. 10,258,641, filed on April 16, 2019; and U.S. Patent No. 10,300,086, filed on May 28, 2019), as well as the compositions described in U.S. Provisional Application No. 63 / 311,661, titled "COMPOSITIONS OF HYDROXYPROPYL-BETA-CYCLODEXTRIN AND METHODS OF PURIFYING THE SAME", filed on February 18, 2022. These patents, patent applications, and provisional applications are each incorporated herein by reference in their entirety.

[0240] Method for producing beta-cyclodextrin The beta-cyclodextrin (BCD) used in the systems and methods described herein can be produced by an enzymatic synthesis process. Suitable enzymatic synthesis processes are disclosed, for example, in PCT / IB2023 / 055977, the disclosure of which is incorporated herein by reference.

[0241] In some cases, a method for producing BCD or a method for producing a composition containing cyclodextrin comprises contacting sucrose with an enzyme or enzyme mixture capable of converting sucrose to amylose under conditions that allow the conversion of sucrose to amylose, thereby producing amylose. In some cases, the method further comprises contacting amylose with an enzyme capable of converting amylose to cyclodextrin under conditions that allow the conversion of amylose to cyclodextrin, thereby producing a composition containing cyclodextrin. In some cases, the enzyme capable of converting amylose to cyclodextrin is a variant enzyme that can produce beta-cyclodextrin in a higher amount and / or concentration (e.g., weight %, mol %, or w / v) than the wild-type enzyme capable of converting amylose to cyclodextrin, alpha-cyclodextrin, gamma-cyclodextrin, or both. In some cases, the composition containing cyclodextrin contains beta-cyclodextrin and may further optionally contain alpha-cyclodextrin, gamma-cyclodextrin, or any combination thereof. In some cases, the composition containing cyclodextrin contains beta-cyclodextrin in a higher amount and / or concentration (e.g., weight %, mol %, or w / v) than alpha-cyclodextrin, gamma-cyclodextrin, or both. In some cases, the amounts and / or concentrations of alpha-cyclodextrin, beta-cyclodextrin, and gamma-cyclodextrin are measured by high performance liquid chromatography (HPLC).

[0242] Method step (a) for the enzymatic conversion from sucrose to amylose The methods provided herein may include the enzymatic conversion of sucrose to amylose. In some cases, the amylose is alpha - amylose. In some embodiments, the method includes contacting sucrose with an enzyme or enzyme mixture capable of converting sucrose to amylose under conditions that allow for the conversion of sucrose to amylose, thereby producing amylose. In one aspect, the method includes the use of a single enzyme for converting sucrose to amylose. In an alternative aspect, the method includes the use of an enzyme mixture (e.g., two enzymes) that collectively or in combination convert sucrose to amylose. In some cases, the sucrose is deuterated sucrose (e.g., one or more hydrogens are replaced with deuterium). In some cases, the sucrose and / or any one or more reagents used in the synthesis reaction are deuterated.

[0243] One-enzyme method for producing amylose from sucrose In some embodiments, the enzyme is amylosucrase. Figure 27A shows a schematic of a single - enzyme method for producing amylose from sucrose. In this example, sucrose is contacted with amylosucrase, which converts the sucrose to amylose. In some cases, the amylosucrase is wild - type amylosucrase. For example, the wild - type amylosucrase can be Cellulomonas carboniz T26 amylosucrase (NCBI accession number N868_11335). In some cases, the wild - type Cellulomonas carboniz T26 amylosucrase can have the amino acid sequence of SEQ ID NO: 1. In some cases, the wild - type amylosucrase can be Neisseria polysaccharea amylosucrase (NCBI accession number AJ011781). In some cases, the wild - type Neisseria polysaccharea amylosucrase can have the amino acid sequence of SEQ ID NO: 2. Table 1 below shows non - limiting examples of wild - type amylosucrase enzymes (and their amino acid sequences) that can be used in accordance with the methods provided herein.

Table 1

[0244] In some embodiments, the amylosucrase is a variant amylosucrase that includes at least one amino acid variant relative to wild-type amylosucrase. The variant amylosucrase can include one or more amino acid substitutions, deletions, insertions, and / or modifications relative to wild-type amylosucrase. Optionally, the variant amylosucrase can produce a higher amount and / or concentration of amylose from sucrose relative to wild-type amylosucrase.

[0245] In some cases, variant amylosucrase comprises at least one amino acid variant relative to wild-type Cellulomonas carboniz T26 amylosucrase (SEQ ID NO: 1). In some cases, variant amylosucrase comprises at least one amino acid variant relative to wild-type Neisseria polysaccharea amylosucrase (SEQ ID NO: 2). In some cases, variant amylosucrase comprises or consists of an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of wild-type Cellulomonas carboniz T26 amylosucrase, preferably at least about 90% sequence identity. In some cases, variant amylosucrase comprises or consists of an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 1, preferably at least about 90% sequence identity.In some cases, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of wild-type Neisseria polysaccharea amylosucrase, preferably at least about 90% sequence identity. In some cases, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity.

[0246] In some cases, at least one amino acid variant contains at least one amino acid substitution relative to the wild-type amylosucrase. In some cases, at least one amino acid variant contains at least one amino acid substitution relative to the wild-type Cellulomonas carboniz T26 amylosucrase. In some cases, at least one amino acid variant contains at least one amino acid substitution relative to the wild-type Neisseria polysaccharea amylosucrase. In some cases, at least one amino acid substitution comprises or consists of an amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2. In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is selected from the group consisting of R234Q, R234G, R234A, R234S, R234M, R234C, R234K, R234I, R234D, R234Y, R234W, R234E, R234L, and R234H. In a preferred embodiment, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is selected from the group consisting of R234Q, R234G, R234A, R234S, R234M, R234C, and R234K. In this regard, it will be understood that R234Q indicates that the arginine (R) at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is replaced by glutamine (Q), and so on. In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is R234Q (e.g., SEQ ID NO: 3 in Table 2). In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is R234G (e.g., SEQ ID NO: 4 in Table 2). In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is R234A (e.g., SEQ ID NO: 5 in Table 2). In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is R234S (e.g., SEQ ID NO: 6 in Table 2). In some cases, the amino acid substitution at amino acid position 234 relative to the amino acid sequence of SEQ ID NO: 2 is R234M (e.g., SEQ ID NO: 7 in Table 2).In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234C (e.g., SEQ ID NO: 8 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234K (e.g., SEQ ID NO: 9 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234I (e.g., SEQ ID NO: 10 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234D (e.g., SEQ ID NO: 11 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234Y (e.g., SEQ ID NO: 12 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234W (e.g., SEQ ID NO: 13 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234E (e.g., SEQ ID NO: 14 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234L (e.g., SEQ ID NO: 15 in Table 2). In some cases, the amino acid substitution at position 234 with respect to the amino acid sequence of SEQ ID NO: 2 is R234H (e.g., SEQ ID NO: 16 in Table 2). In some embodiments, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity to the amino acid sequence according to any one of SEQ ID NOs: 3 to 16 or 48 shown in Table 2, or the amino acid sequence according to any one of SEQ ID NOs: 3 to 16 or 48 shown in Table 2.In a preferred embodiment, the variant amylase comprises, or consists of, an amino acid sequence according to any one of SEQ ID NOs: 3 to 9 or 48 shown in Table 2. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0247] In some embodiments, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 234 relative to SEQ ID NO:2. In this regard, as used throughout this disclosure, the described sequence identity includes amino acid substitutions (i.e., the sequence identity is calculated based on the entire amino acid sequence of the variant enzyme including the amino acid substitutions). Optionally, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 234 relative to SEQ ID NO:2 selected from the group consisting of R234Q, R234G, R234A, R234S, R234M, R234C, R234K, R234I, R234D, R234Y, R234W, R234E, R234L, and R234H.In a preferred embodiment, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 2, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 234 of SEQ ID NO: 2 selected from the group consisting of R234Q, R234G, R234A, R234S, R234M, R234C, and R234K. Optionally, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 2, preferably at least about 90% sequence identity, and an amino acid substitution R234Q of SEQ ID NO: 2. Optionally, the variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 2, preferably at least about 90% sequence identity, and an amino acid substitution R234G of SEQ ID NO: 2.In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234A relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234S relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234M relative to SEQ ID NO:2.In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234C relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234K relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234I relative to SEQ ID NO:2.In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234D relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234Y relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234W relative to SEQ ID NO:2.In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234E relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234L relative to SEQ ID NO:2. In some cases, variant amylosucrase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO:2, preferably at least about 90% sequence identity, and the amino acid substitution R234H relative to SEQ ID NO:2.

[0248] In some embodiments, the amylosucrase is derived from microbial cells. Optionally, the amylosucrase is isolated and / or purified from microbial cells. Optionally, the microbial cells are bacterial cells. Optionally, the bacterial cells are Escherichia coli. In some embodiments, the amylosucrase is derived from Neisseria polysaccharea. In some embodiments, the amylosucrase is derived from Cellulomonas carboniz T26. In some embodiments, the amylosucrase can be produced within microbial cells. In some embodiments, the amylosucrase is expressed within recombinant host cells (e.g., from a recombinant polynucleotide). Optionally, the amylosucrase is produced recombinantly. Optionally, the amylosucrase is produced in yeast cells (e.g., produced recombinantly). Optionally, the yeast cells are Pichia yeast cells such as Pichia pastoris cells.

[0249] Two-enzyme method for producing amylose from sucrose In some embodiments, the method includes contacting sucrose with an enzyme mixture capable of converting sucrose to amylose under conditions that allow for the conversion of sucrose to amylose, thereby producing amylose. Optionally, the method includes contacting sucrose with an enzyme mixture that includes at least two enzymes capable of collectively or combinatorially converting sucrose to amylose. For example, the enzyme mixture can include at least sucrose phosphorylase and alpha-glucan phosphorylase. The method can include contacting sucrose with at least two enzymes simultaneously or substantially simultaneously. Alternatively, the method can include contacting sucrose with at least two enzymes sequentially. FIG. 27B shows a schematic of a two-enzyme method for producing amylose from sucrose. In this example, sucrose is contacted with sucrose phosphorylase to convert sucrose to glucose-1-phosphate. Next, glucose-1-phosphate is contacted with alpha-glucan phosphorylase to convert glucose-1-phosphate to amylose. Optionally, sucrose phosphorylase and alpha-glucan phosphorylase are contacted with sucrose simultaneously or substantially simultaneously. In other cases, sucrose phosphorylase and alpha-glucan phosphorylase are added sequentially (e.g., sucrose phosphorylase is first contacted with sucrose to produce glucose-1-phosphate, and then alpha-glucan phosphorylase is added to produce amylose). Optionally, glucose-1-phosphate generated from the reaction with sucrose phosphorylase is isolated and / or purified before contacting it with alpha-glucan phosphorylase. In other cases, glucose-1-phosphate generated from the reaction with sucrose phosphorylase is not isolated and / or purified before contacting it with alpha-glucan phosphorylase. As used in the context of adding two or more components to the reaction mixture described herein, the term "substantially simultaneously" means that the two or more components are added to the reaction mixture within 10 seconds of each other.

[0250] In some cases, the sucrose phosphorylase is a wild-type sucrose phosphorylase. For example, the wild-type sucrose phosphorylase can be Bifidobacterium longum sucrose phosphorylase (e.g., NCBI accession number AAO84039). In some cases, the wild-type Bifidobacterium longum sucrose phosphorylase can have the amino acid sequence according to SEQ ID NO: 17. In some cases, the wild-type sucrose phosphorylase can be Leuconostoc mesenteroide sucrose phosphorylase (e.g., NCBI accession number D90314.1). In some cases, the wild-type Leuconostoc mesenteroide sucrose phosphorylase can have the amino acid sequence according to SEQ ID NO: 18. In some cases, the wild-type sucrose phosphorylase can be Streptococcus mutans sucrose phosphorylase (e.g., NCBI accession number NZ_CP013237.1). In some cases, the wild-type Streptococcus mutans sucrose phosphorylase can have the amino acid sequence according to SEQ ID NO: 19 (e.g., NCBI accession number P10249). In some cases, the sucrose phosphorylase enzyme is a variant sucrose phosphorylase enzyme. In some cases, the variant sucrose phosphorylase has one or more amino acid substitutions relative to the wild-type sucrose phosphorylase. In some cases, the variant sucrose phosphorylase has amino acid substitutions in one or all of amino acid residues T47, S62, Y77, V128, K140, Q144, N155, and D249 relative to SEQ ID NO: 19. In some cases, the amino acid substitution at amino acid position 47 relative to SEQ ID NO: 19 is T47S. In some cases, the amino acid substitution at amino acid position 62 relative to SEQ ID NO: 19 is S62P. In some cases, the amino acid substitution at amino acid position 77 relative to SEQ ID NO: 19 is Y77H. In some cases, the amino acid substitution at amino acid position 128 relative to SEQ ID NO: 19 is V128L. In some cases, the amino acid substitution at amino acid position 140 relative to SEQ ID NO: 19 is K140M.In some cases, the amino acid substitution at position 144 for SEQ ID NO: 19 is Q144R. In some cases, the amino acid substitution at position 155 for SEQ ID NO: 19 is N155S. In some cases, the amino acid substitution at position 249 for SEQ ID NO: 19 is D249G. In some cases, the variant sucrose phosphorylase has the amino acid substitutions T47S, S62P, Y77H, V128L, K140M, Q144R, N155S, and D249G with respect to SEQ ID NO: 19. In some cases, the variant sucrose phosphorylase comprises or consists of the amino acid sequence according to SEQ ID NO: 20. Table 3 below shows non-limiting examples of sucrose phosphorylase enzymes (and their amino acid sequences) that can be used according to the methods provided herein.

Table 3-1

Table 3-2

[0251] In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild-type Bifidobacterium longum sucrose phosphorylase, preferably at least about 90% sequence identity. In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 17, preferably at least about 90% sequence identity. In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild-type Leuconostoc mesenteroides sucrose phosphorylase, preferably at least about 90% sequence identity.In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 18, preferably at least about 90% sequence identity. In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild-type Streptococcus mutans sucrose phosphorylase, preferably at least about 90% sequence identity. In some cases, the sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 19, preferably at least about 90% sequence identity.In some cases, sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 20, preferably at least about 90% sequence identity, and comprises the amino acid substitutions T47S, S62P, Y77H, V128L, K140M, Q144R, N155S, and D249G relative to SEQ ID NO: 19.

[0252] In some embodiments, sucrose phosphorylase is derived from a microbial cell. In some cases, sucrose phosphorylase is isolated and / or purified from a microbial cell. In some cases, the microbial cell is a bacterial cell. In some cases, the bacterial cell is Escherichia coli. In some embodiments, sucrose phosphorylase is derived from Bifidobacterium longum. In some embodiments, sucrose phosphorylase is derived from Leuconostoc mesenteroides. In some embodiments, sucrose phosphorylase is derived from Streptococcus mutans. In some embodiments, sucrose phosphorylase can be produced within a microbial cell. In some embodiments, sucrose phosphorylase is expressed within a recombinant host cell (e.g., from a recombinant polynucleotide). In some cases, sucrose phosphorylase is produced recombinantly. In some cases, sucrose phosphorylase is produced in a yeast cell (e.g., produced recombinantly). In some cases, the yeast cell is a Pichia yeast cell such as a Pichia pastoris cell.

[0253] In some embodiments, the alpha-glucan phosphorylase is a wild-type alpha-glucan phosphorylase. In some cases, the wild-type alpha-glucan phosphorylase can be a Solanum tuberosum alpha-glucan phosphorylase (e.g., NCBI accession number D00520.1). In some cases, the wild-type Solanum tuberosum alpha-glucan phosphorylase can have the amino acid sequence according to SEQ ID NO: 21. In some cases, the wild-type alpha-glucan phosphorylase can be an S. tokodaii strain 7 alpha-glucan phosphorylase (e.g., NCBI accession number NC_003106.2). In some cases, the wild-type S. tokodaii strain 7 alpha-glucan phosphorylase can have the amino acid sequence according to SEQ ID NO: 22. In some cases, the wild-type alpha-glucan phosphorylase can be a C. callunae DSM 20145 alpha-glucan phosphorylase (e.g., NCBI accession number AY102616.1). In some cases, the wild-type C. callunae DSM 20145 alpha-glucan phosphorylase can have the amino acid sequence according to SEQ ID NO: 23. In some cases, the alpha-glucan phosphorylase enzyme is a variant alpha-glucan phosphorylase enzyme. In some cases, the variant alpha-glucan phosphorylase has one or more amino acid substitutions relative to the wild-type alpha-glucan phosphorylase. In some cases, the variant alpha-glucan phosphorylase has an amino acid substitution in one or all of amino acid residues F39, N135, and T706 relative to SEQ ID NO: 21. In some cases, the amino acid substitution at amino acid position 39 relative to SEQ ID NO: 21 is F39L. In some cases, the amino acid substitution at amino acid position 135 relative to SEQ ID NO: 21 is N135S. In some cases, the amino acid substitution at amino acid position 706 relative to SEQ ID NO: 21 is T706I. In some cases, the variant alpha-glucan phosphorylase has the amino acid substitutions F39L, N135S, and T706I relative to SEQ ID NO: 21.In some cases, the variant alpha - glucan phosphorylase enzyme comprises or consists of the amino acid sequence according to SEQ ID NO: 24. Table 4 below shows non - limiting examples of alpha - glucan phosphorylase enzymes (and their amino acid sequences) that can be used in accordance with the methods provided herein.

Table 4 - 1

Table 4 - 2

[0254] In some cases, the alpha - glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild - type Solanum tuberosum alpha - glucan phosphorylase, preferably at least about 90% sequence identity. In some cases, the alpha - glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 21, preferably at least about 90% sequence identity. In some cases, the alpha - glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild - type S. tokodaii strain 7 alpha - glucan phosphorylase, preferably at least about 90% sequence identity.In some cases, the alpha-glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 22, preferably at least about 90% sequence identity. In some cases, the alpha-glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the wild-type C. callunae DSM 20145 alpha-glucan phosphorylase, preferably at least about 90% sequence identity. In some cases, the alpha-glucan phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 23, preferably at least about 90% sequence identity.In some cases, sucrose phosphorylase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 24, preferably at least about 90% sequence identity, and includes the amino acid substitutions F39L, N135S, and T706I relative to SEQ ID NO: 21.

[0255] In some embodiments, the alpha - glucan phosphorylase is derived from microbial cells. In some cases, the alpha - glucan phosphorylase is isolated and / or purified from microbial cells. In some cases, the microbial cells are bacterial cells. In some cases, the bacterial cells are Escherichia coli. In some embodiments, the alpha - glucan phosphorylase is derived from Solanum tuberosum. In some embodiments, the alpha - glucan phosphorylase is derived from the S. tokodaii strain 7. In some embodiments, the alpha - glucan phosphorylase is derived from C. callunae DSM 20145. In some embodiments, the alpha - glucan phosphorylase can be produced within microbial cells. In some embodiments, the alpha - glucan phosphorylase is expressed within recombinant host cells (e.g., from a recombinant polynucleotide). In some cases, the alpha - glucan phosphorylase is produced recombinantly. In some cases, the alpha - glucan phosphorylase is produced in yeast cells (e.g., produced recombinantly). In some cases, the yeast cells are Pichia yeast cells such as Pichia pastoris cells.

[0256] Method step (b) for the enzymatic conversion from amylose to beta-cyclodextrin In various embodiments, the method further comprises enzymatically converting amylose (e.g., as produced by the methods provided herein (e.g., method step (a))) to cyclodextrin, preferably beta-cyclodextrin. Optionally, the method comprises contacting amylose with an enzyme or enzyme mixture (e.g., two or more enzymes, etc.) capable of converting amylose to cyclodextrin under conditions that allow for the conversion of amylose to cyclodextrin. Optionally, the enzyme capable of converting amylose to cyclodextrin is a variant enzyme that can produce a higher amount and / or concentration of beta-cyclodextrin than alpha-cyclodextrin, gamma-cyclodextrin, or both, relative to a wild-type enzyme capable of converting amylose to cyclodextrin.

[0257] In some embodiments, the enzyme capable of converting amylose to cyclodextrin comprises a variant cyclodextrin glucanotransferase. Optionally, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to a wild-type cyclodextrin glucanotransferase. Figure 28 shows the enzymatic conversion of amylose to beta-cyclodextrin using cyclodextrin glucanotransferase. Preferably, the cyclodextrin glucanotransferase produces beta-cyclodextrin from amylose in a higher amount and / or concentration than the amount and / or concentration of alpha-cyclodextrin and / or gamma-cyclodextrin.

[0258] In some embodiments, the cyclodextrin glucanotransferase is a variant cyclodextrin glucanotransferase comprising at least one amino acid variant relative to the wild-type cyclodextrin glucanotransferase. The variant cyclodextrin glucanotransferase can include one or more amino acid substitutions, deletions, insertions, and / or modifications relative to the wild-type cyclodextrin glucanotransferase. Optionally, the variant cyclodextrin glucanotransferase can produce a higher amount and / or concentration of beta-cyclodextrin from amylose compared to alpha-cyclodextrin and / or gamma-cyclodextrin relative to the wild-type cyclodextrin glucanotransferase.

[0259] In some cases, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to the wild-type Bacillus sp. (strain number 38-2) cyclodextrin glucanotransferase (e.g., NCBI accession number M19880.1; SEQ ID NO: 25). In some cases, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to the wild-type B. circulans strain 251 cyclodextrin glucanotransferase (e.g., NCBI accession number X78145.1; SEQ ID NO: 26 or 27). In some cases, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to the wild-type B. circulans strain 251 cyclodextrin glucanotransferase of SEQ ID NO: 27. In some cases, the variant cyclodextrin glucanotransferase comprises or consists of an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 25, preferably at least about 90% sequence identity. In some cases, the variant cyclodextrin glucanotransferase comprises or consists of an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 26 or 27, preferably at least about 90% sequence identity.In some cases, variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 27, preferably at least about 90% sequence identity.

[0260] In some cases, at least one amino acid variant comprises at least one amino acid substitution relative to wild-type cyclodextrin glucanotransferase. In some cases, at least one amino acid substitution comprises an amino acid substitution at amino acid position 31 relative to the amino acid sequence of SEQ ID NO: 27. In some cases, the amino acid substitution at amino acid position 31 relative to the amino acid sequence of SEQ ID NO: 27 is A31R (e.g., SEQ ID NO: 28 in Table 5). In some cases, the amino acid substitution at amino acid position 31 relative to the amino acid sequence of SEQ ID NO: 27 is A31P (e.g., SEQ ID NO: 29 in Table 5). In some cases, the amino acid substitution at amino acid position 31 relative to the amino acid sequence of SEQ ID NO: 27 is A31T (e.g., SEQ ID NO: 30 in Table 5). In some embodiments, cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence according to any one of SEQ ID NOS: 25-30 shown in Table 5.

[0261] In some cases, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to wild-type Paenibacillus macerans cyclodextrin glucanotransferase (e.g., NCBI accession number AAA22298.1 or X59045.1; e.g., SEQ ID NOs: 31-34). In some cases, the variant cyclodextrin glucanotransferase comprises at least one amino acid variant relative to any one of SEQ ID NOs: 31-34. In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) to the amino acid sequence of wild-type Paenibacillus macerans cyclodextrin glucanotransferase, preferably at least about 90% sequence identity. In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) to the amino acid sequence of any one of SEQ ID NOs: 31-34, preferably at least about 90% sequence identity.

[0262] In some cases, at least one amino acid variant comprises at least one amino acid substitution relative to wild-type cyclodextrin glucanotransferase. In some cases, at least one amino acid substitution comprises an amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34. In some cases, the amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34 is R146A (e.g., SEQ ID NO: 35 in Table 5). In some cases, the amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34 is R146P (e.g., SEQ ID NO: 36 in Table 5). In some cases, at least one amino acid substitution comprises an amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34. In some cases, the amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34 is D147A (e.g., SEQ ID NO: 37 in Table 5). In some cases, the amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34 is D147P (e.g., SEQ ID NO: 38 in Table 5). In some cases, at least one amino acid substitution comprises amino acid substitutions at amino acid positions 146 and 147 relative to the amino acid sequence of SEQ ID NO: 34. In some cases, the amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34 is R146A and the amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34 is D147P (e.g., SEQ ID NO: 39 in Table 5). In some cases, the amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34 is R146P and the amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34 is D147A (e.g., SEQ ID NO: 40 in Table 5). In some cases, the amino acid substitution at amino acid position 146 relative to the amino acid sequence of SEQ ID NO: 34 is R146P and the amino acid substitution at amino acid position 147 relative to the amino acid sequence of SEQ ID NO: 34 is D147P (e.g., SEQ ID NO: 41 in Table 5).

[0263] In some cases, at least one amino acid substitution includes an amino acid substitution at position 372 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34. In some cases, the amino acid substitution at position 372 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34 is D372K (for example, SEQ ID NO: 42 (for SEQ ID NO: 32) and SEQ ID NO: 45 (for SEQ ID NO: 34) in Table 5). In some cases, at least one amino acid substitution includes an amino acid substitution at position 89 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34. In some cases, the amino acid substitution at position 89 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34 is Y89R (for example, SEQ ID NO: 43 (for SEQ ID NO: 32) and SEQ ID NO: 47 (for SEQ ID NO: 34) in Table 5). In some cases, at least one amino acid substitution includes an amino acid substitution at position 372 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34 and an amino acid substitution at position 89 with respect to the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 34. In some cases, the amino acid substitution at position 372 with respect to the amino acid sequence of SEQ ID NO: 32 or 34 is D372K, and the amino acid substitution at position 89 with respect to the amino acid sequence of SEQ ID NO: 32 or 34 is Y89R (for example, SEQ ID NO: 44 (for SEQ ID NO: 32) and SEQ ID NO: 47 (for SEQ ID NO: 34) in Table 5).

[0264] In some embodiments, cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence according to any one of SEQ ID NOs: 31 to 47 shown in Table 5. In some embodiments, cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity, preferably at least about 90% sequence identity, with an amino acid sequence according to any one of SEQ ID NOs: 31 to 47 shown in Table 5.

[0265] In certain embodiments, cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence according to SEQ ID NO: 34, or comprises, or consists of, an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity, preferably at least about 90% sequence identity, with the amino acid sequence according to SEQ ID NO: 34.

[0266] In another specific embodiment, the cyclodextrin glucanotransferase comprises or consists of the amino acid sequence according to SEQ ID NO: 39, or comprises or consists of an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity with the amino acid sequence according to SEQ ID NO: 39, preferably at least about 90% sequence identity.

[0267] In another specific embodiment, the cyclodextrin glucanotransferase comprises or consists of the amino acid sequence according to SEQ ID NO: 40, or comprises or consists of an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity with the amino acid sequence according to SEQ ID NO: 40, preferably at least about 90% sequence identity.

[0268] In another specific embodiment, the cyclodextrin glucanotransferase comprises, or consists of, the amino acid sequence according to SEQ ID NO: 41, or an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity with the amino acid sequence according to SEQ ID NO: 41, preferably at least about 90% sequence identity.

[0269] In another specific embodiment, the cyclodextrin glucanotransferase comprises, or consists of, the amino acid sequence according to SEQ ID NO: 47, or an amino acid sequence having at least about 70% (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) sequence identity with the amino acid sequence according to SEQ ID NO: 47, preferably at least about 90% sequence identity.

Table 5-1

Table 5-2

Table 5-3

Table 5-4

Table 5-5

Table 5-6

Table 5-7

Table 5-8

Table 5-9

Table 5-10

[0270] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 25, preferably at least about 90% sequence identity. In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 26 or 27, preferably at least about 90% sequence identity.

[0271] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 27, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 31 relative to SEQ ID NO: 27. In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 27, preferably at least about 90% sequence identity, and an amino acid substitution A31R at amino acid position 31 relative to SEQ ID NO: 27. In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 27, preferably at least about 90% sequence identity, and an amino acid substitution A31P at amino acid position 31 relative to SEQ ID NO: 27.In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 27, preferably at least about 90% sequence identity, and the amino acid substitution A31T relative to SEQ ID NO: 27.

[0272] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 146 relative to SEQ ID NO: 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and the amino acid substitution R146A relative to SEQ ID NO: 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and the amino acid substitution R146P relative to SEQ ID NO: 34.

[0273] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 147 relative to SEQ ID NO: 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and an amino acid substitution D147P relative to SEQ ID NO: 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, and an amino acid substitution D147A relative to SEQ ID NO: 34.

[0274] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, an amino acid substitution at amino acid position 146 relative to SEQ ID NO: 34, and an amino acid substitution at amino acid position 147 relative to SEQ ID NO: 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, the amino acid substitution R146A relative to SEQ ID NO: 34, and the amino acid substitution D147P relative to SEQ ID NO: 34.In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, an amino acid substitution R146P with respect to SEQ ID NO: 34, and an amino acid substitution D147A with respect to SEQ ID NO: 34. In some cases, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 34, preferably at least about 90% sequence identity, an amino acid substitution R146P with respect to SEQ ID NO: 34, and an amino acid substitution D147P with respect to SEQ ID NO: 34.

[0275] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 372 relative to SEQ ID NO: 32 or 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, and an amino acid sequence having the amino acid substitution D372K relative to SEQ ID NO: 32 or 34.

[0276] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, and an amino acid substitution at amino acid position 89 relative to SEQ ID NO: 32 or 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, and an amino acid substitution Y89R relative to SEQ ID NO: 32 or 34.

[0277] In some embodiments, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence identity (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, an amino acid substitution at amino acid position 372 relative to SEQ ID NO: 32 or 34, and an amino acid substitution at amino acid position 89 relative to SEQ ID NO: 32 or 34. Optionally, the variant cyclodextrin glucanotransferase comprises, or consists of, an amino acid sequence having at least about 70% sequence (e.g., at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more) with the amino acid sequence of SEQ ID NO: 32 or 34, preferably at least about 90% sequence identity, the amino acid substitution D372K relative to SEQ ID NO: 32 or 34, and the amino acid substitution Y89R relative to SEQ ID NO: 32 or 34.

[0278] In some embodiments, cyclodextrin glucanotransferase is derived from microbial cells. Optionally, cyclodextrin glucanotransferase is isolated and / or purified from microbial cells. Optionally, the microbial cells are bacterial cells. Optionally, the bacterial cells are Escherichia coli. In some embodiments, cyclodextrin glucanotransferase is derived from Bacillus sp. (strain number 38-2). In some embodiments, cyclodextrin glucanotransferase is derived from B. circulans strain 251. In some embodiments, cyclodextrin glucanotransferase can be produced within microbial cells. In some embodiments, cyclodextrin glucanotransferase is expressed within recombinant host cells (e.g., from a recombinant polynucleotide). Optionally, cyclodextrin glucanotransferase is produced recombinantly. Optionally, cyclodextrin glucanotransferase is produced in yeast cells (e.g., produced recombinantly). Optionally, the yeast cells are Pichia yeast cells such as Pichia pastoris cells.

[0279] In various embodiments, the methods provided herein produce beta-cyclodextrin at a higher ratio than alpha-cyclodextrin, gamma-cyclodextrin, or both. For example, in some cases, the methods provided herein provide a ratio of beta-cyclodextrin to alpha-cyclodextrin, gamma-cyclodextrin, or both of at least 2:1, at least 3:1, at least 4:1, at least 5:1, at least 6:1, at least 7:1, at least 8:1, at least 9:1, at least 10:1, at least 20:1, at least 30:1, at least 40:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, at least 100:1, or more. In a preferred embodiment, the methods provided herein provide a ratio of beta-cyclodextrin to alpha-cyclodextrin of at least 10:1. For example, the ratio can be at least 20:1, at least 30:1, at least 40:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, at least 100:1, or more. In a preferred embodiment, the methods provided herein provide a ratio of beta-cyclodextrin to gamma-cyclodextrin of at least 5:1. For example, the ratio can be at least 10:1, at least 20:1, at least 30:1, at least 40:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, at least 100:1, or more. In a preferred embodiment, the methods provided herein provide a ratio of beta-cyclodextrin to both alpha-cyclodextrin and gamma-cyclodextrin of at least 3.5:1. For example, the ratio can be at least 5:1, at least 10:1, at least 20:1, at least 30:1, at least 40:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, at least 100:1, or more.

[0280] Throughout the present disclosure, methods are outlined for achieving robust enzyme activity at each step to obtain a higher yield of beta-cyclodextrin than is currently achievable. In some embodiments, a first enzymatic step of converting sucrose to amylose (e.g., as described herein) is carried out over a first period, thereby enabling a catalytic conversion of sucrose to amylose, followed by a second enzymatic step of converting amylose to beta-cyclodextrin (e.g., as described herein) over a second period, thereby enabling a catalytic conversion of amylose to beta-cyclodextrin. In some embodiments, the first enzymatic reaction (e.g., as described herein, e.g., converting sucrose to amylose) and the second enzymatic reaction (e.g., as described herein, e.g., converting amylose to beta-cyclodextrin) are carried out in the same reservoir (e.g., one-pot synthesis method).

[0281] In some embodiments, the first period is at least 30 minutes, at least 45 minutes, at least 60 minutes, at least 85 minutes, at least 90 minutes, at least 105 minutes, at least 120 minutes, at least 135 minutes, at least 150 minutes, at least 165 minutes, at least 180 minutes, at least 195 minutes, at least 210 minutes, at least 225 minutes, at least 240 minutes, at least 255 minutes, at least 270 minutes, at least 285 minutes, or at least 300 minutes. In some embodiments, the second period is at least 30 minutes, at least 45 minutes, at least 60 minutes, at least 85 minutes, at least 90 minutes, at least 105 minutes, at least 120 minutes, at least 135 minutes, at least 150 minutes, at least 165 minutes, at least 180 minutes, at least 195 minutes, at least 210 minutes, at least 225 minutes, at least 240 minutes, at least 255 minutes, at least 270 minutes, at least 285 minutes, or at least 300 minutes. In some embodiments, the first period is shorter than the second period. In some embodiments, the first period is longer than the second period. In some embodiments, the first period is the same or substantially the same length as the second period. In some embodiments, sucrose is added to the reaction reservoir batch by batch. In some embodiments, the enzyme used in the first enzyme reaction step (e.g., as described herein, e.g., to convert sucrose to amylose) is added once at the start of the reaction period and then added again after a certain period of time has elapsed to promote catalytic activity. In some embodiments, sucrose is added once at the start of the reaction period and then added again after a certain period of time has elapsed to replenish the sucrose. In some embodiments, the enzyme used in the first enzyme reaction step (e.g., as described herein, e.g., to convert sucrose to amylose) is added to the same reaction reservoir at the same time as the enzyme used in the second enzyme reaction step (e.g., to convert amylose to beta-cyclodextrin).In some embodiments, the enzyme used in the first enzyme reaction step (e.g., as described herein, e.g., to convert sucrose to amylose) is added at a different time point (e.g., prior thereto) than the enzyme used in the second enzyme reaction step (e.g., to convert amylose to beta-cyclodextrin).

[0282] In some embodiments, the sucrose concentration is maximized for efficient conversion to amylose. In some embodiments, the starting concentration of sucrose in the reaction is at least about 50 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 100 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 150 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 200 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 250 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 300 g / L. In some embodiments, the starting concentration of sucrose in the reaction is at least about 350 g / L.

[0283] In some embodiments, the reaction time is an important consideration for obtaining beta-cyclodextrin at maximum yield. In some embodiments, the production of beta-cyclodextrin can involve the decomposition of the product into glucose, maltose, and other sugars. Therefore, it is important to obtain beta-cyclodextrin without decomposing it. In some embodiments, the entire reaction (e.g., method step (a) and method step (b)) is carried out over 12 hours or less. In some embodiments, the entire reaction (e.g., method step (a) and method step (b)) is carried out over 8 hours or less. In some embodiments, the entire reaction is carried out over 7 hours or less. In some embodiments, the entire reaction is carried out over 6 hours or less. In some embodiments, the entire reaction is carried out over 5 hours or less. In some embodiments, the entire reaction is carried out over 4 hours or less. In some embodiments, the entire reaction is carried out over 3 hours or less. In some embodiments, the entire reaction is carried out over 2 hours or less. In some embodiments, the entire reaction is carried out over 1 hour or less.

[0284] Temperature is an important consideration for maximizing the yield of beta-cyclodextrin. In some embodiments, one or more of the enzymatic reactions are carried out at about 30°C to about 55°C, such as about 40°C to about 50°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 40°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 41°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 42°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 43°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 44°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 45°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 46°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 47°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 48°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 49°C. In some embodiments, one or more of the enzymatic reactions are carried out at about 50°C. Preferably, one or more of the reactions are carried out at about 45°C.

[0285] In some embodiments, the enzymatic reaction of step (a) is carried out at about 40°C to about 55°C, such as about 45°C to about 50°C. In some embodiments, the enzymatic reaction of step (b) is carried out at about 40°C to about 50°C. Step (a) and step (b) may be carried out at different temperatures, or preferably, step (a) and step (b) are carried out at substantially the same temperature. If step (a) involves the use of a single enzyme (e.g., amylosucrase), the enzymatic reaction of step (a) is preferably carried out at about 45°C. In this embodiment, the enzymatic reaction of step (b) is preferably also carried out at about 45°C. If step (a) involves the use of at least two enzymes (e.g., sucrose phosphorylase and alpha-glucan phosphorylase), the enzymatic reaction of step (a) is preferably carried out at about 45°C or about 50°C. In this embodiment, the enzymatic reaction of step (b) is preferably also carried out at about 45°C or about 50°C, respectively.

[0286] In one-pot synthesis, even though the optimal temperature for each enzyme may vary slightly, it is considered that the functionality of the enzyme mixture(s) should be maximized.

[0287] In some embodiments, the reaction is carried out in a reservoir having a reservoir volume of about 1 mL to about 1,000,000 L. For example, the reaction can be carried out in a reservoir having a reservoir volume of about 100 mL to about 10 L, such as a reservoir volume of about 500 mL or about 10 L.

[0288] In some embodiments, the total reaction volume is from about 1 mL to about 1,000,000 L. For example, the total reaction volume can be from about 100 mL to about 10 L, such as a total reaction volume of about 500 mL or about 5 L. In some embodiments, the total reaction volume is smaller than the reservoir volume. For example, in a reaction carried out in a reservoir having a reservoir volume of about 10 L, a total reaction volume of about 5 L can be used.

[0289] In some embodiments, the reaction is carried out in a stirred tank reactor (STR), a loop reactor, a plug flow reactor, a single-stage or multi-stage continuous stirred tank reactor, or any other suitable reactor known in the art. In some embodiments, the reaction is carried out in a stirred tank reactor and the reactants are stirred at about 100 to about 200 rpm, such as about 160 rpm.

[0290] The pH of the reaction mixture can be an important consideration for maximizing the yield of beta-cyclodextrin. In some embodiments, one or more of the enzyme reactions are carried out at a pH of about 6 to about 8, for example, the pH can be from about 6.5 to about 7.5. In preferred embodiments, one or more of the enzyme reactions are carried out at a pH of about 7.0 to about 7.5. Preferably, step (a) is carried out at a pH of about 7.0 to about 7.5. Preferably, step (b) is carried out at a pH of about 7.0 to about 7.5. Steps (a) and (b) may be carried out at different pHs, but preferably, steps (a) and (b) are carried out at the same pH.

[0291] In some embodiments, one or more of the enzymatic reactions are carried out in a reaction mixture containing a buffer. Any suitable buffer known in the art may be used. For example, the buffer may be selected from the group consisting of sodium citrate, disodium hydrogen phosphate, and Tris-HCl. The buffer may be present in the reaction mixture at a concentration of about 50 mM to about 200 mM, such as about 100 mM.

[0292] In some embodiments, one or more of the enzymatic reactions are carried out in a reaction mixture containing an organic solvent, preferably toluene. The reaction mixture preferably also contains water. Without wishing to be bound by any theory set forth herein, the inventors have confirmed that the addition of the organic solvent surprisingly increases the yield of beta-cyclodextrin obtained from the enzymatic reaction. For example, the addition of the organic solvent can increase the yield of beta-cyclodextrin by at least about 5%, such as at least about 10%, such as at least about 15%, such as at least about 20%, such as at least about 50%, such as at least about 100%, such as at least about 150%, such as at least about 200%, such as at least about 250%, such as at least about 300%, such as at least about 350%, such as at least about 400% compared to the yield obtained from the enzymatic reaction carried out without the organic solvent. It is believed that the addition of the organic solvent reduces the solubility of beta-cyclodextrin in the reaction mixture, causing beta-cyclodextrin to precipitate and reducing the concentration of beta-cyclodextrin in the reaction mixture, thereby increasing the yield of beta-cyclodextrin. This prevents the degradation of beta-cyclodextrin by the enzyme.

[0293] In some embodiments, the amount of the organic solvent (preferably toluene) in the reaction mixture is about 0.1 v / v% to about 40 v / v% of the reaction mixture, such as about 1 v / v% to about 35 v / v%, such as about 5 v / v% to about 25 v / v%.

[0294] In some embodiments, the organic solvent is introduced at the start or during the enzymatic reaction of step (a). In some preferred embodiments, the organic solvent is introduced at the start or during the enzymatic reaction of step (b). For example, in embodiments where the entire reaction (e.g., method step (a) and method step (b)) is carried out over 8 hours or less, the organic solvent can be introduced about 1 hour after the start of enzymatic reaction (b).

[0295] In some embodiments, the enzyme used in step (a) is amylosucrase. In some embodiments, the starting concentration of amylosucrase in the reaction mixture is about 1 to about 30 U / mL, such as about 5 to about 25 U / mL, such as about 8 to about 25 U / mL.

[0296] In some embodiments, the enzyme mixture used in step (a) comprises sucrose phosphorylase and alpha - glucan phosphorylase. In some embodiments, the starting concentration of sucrose phosphorylase in the reaction mixture is about 1 to about 30 U / mL, such as about 5 to about 25 U / mL, such as about 8 to about 25 U / mL. In some embodiments, the starting concentration of alpha - glucan phosphorylase in the reaction mixture is about 1 to about 30 U / mL, such as about 5 to about 25 U / mL, such as about 8 to about 25 U / mL.

[0297] In some embodiments, the enzyme is provided in a whole cell lysate, and preferably, the ratio of the starting concentration of the enzyme in step (b) (measured as the volume in the whole cell lysate) to the enzyme in step (a) is about 1:1 to about 50:1, such as about 2:1 to about 50:1, such as about 5:1 to about 40:1, such as about 10:1 to about 30:1. In a preferred embodiment, the ratio is about 20:1.

[0298] In certain embodiments, any one of the enzyme reactions provided herein (e.g., the first enzyme reaction that converts sucrose to amylose and / or the second enzyme reaction that converts amylose to beta-cyclodextrin) can occur within a microbial host cell. Optionally, the microbial cell is a bacterial cell. Optionally, the bacterial cell is Escherichia coli. For example, the microbial host cell can contain one or more heterologous nucleic acid molecules encoding one or more of the enzymes provided herein. The microbial host cell can express one or more of the enzymes provided herein. Optionally, sucrose and / or one or more intermediates of the enzyme reaction can be fed to the microbial host cell. For example, sucrose can be fed to the microbial host cell, and the conversion from sucrose to beta-cyclodextrin can occur within the microbial host cell.

[0299] In some embodiments, one or more of the enzymes used in the enzyme reactions provided herein may be immobilized on a resin. For example, the enzyme may be covalently bound to the resin. Alternatively, the enzyme may be non-covalently bound to the resin. For example, the enzyme may be linked to Ni resin via a His tag. For example, the enzyme of (a) can be a variant amylosucrase (e.g., this variant amylosucrase can contain or consist of the amino acid sequence according to SEQ ID NO: 3), and the enzyme may be immobilized on a resin. Alternatively, or additionally, the enzyme of (b) can be a variant cyclodextrin glucanotransferase (e.g., this variant cyclodextrin glucanotransferase can contain or consist of the amino acid sequence according to SEQ ID NO: 28), and the enzyme may be immobilized on a resin. Optionally, the enzyme or enzyme mixture of (a) and the enzyme of (b) are immobilized on the same resin.

[0300] The resin-immobilized enzyme can be reused by the method described in this specification. However, the inventors have found that when the resin-immobilized enzyme is reused, the yield of beta-cyclodextrin tends to decrease. This is presumably due to the fact that the enzyme leaks from the resin during use, resulting in a decrease in the enzyme conversion rate. Therefore, it is desirable to improve the enzyme stability on the resin and thereby prevent enzyme leakage. This is because it enables the resin-immobilized enzyme to be reused more frequently and / or at a higher enzyme conversion rate, thereby increasing the yield of the reaction.

[0301] The inventors have found that enzyme stability can be improved by using freeze-dried enzymes, spray-drying the enzymes, and / or introducing additives.

[0302] In some embodiments, the enzyme is provided in a cell slurry or in a whole cell lysate. For example, a cell slurry containing recombinant cells expressing the enzyme can be suspended and lysed in a buffer (such as sodium citrate buffer) and centrifuged to provide a whole cell lysate containing the enzyme. Methods of cell lysis are known in the art. For example, cells can be lysed by homogenization, chemical lysis, sonication, freeze / thaw, lytic enzymes, acid lysis, and / or alkaline lysis. In a preferred embodiment, the cells are lysed by homogenization.

[0303] In some embodiments, the cell slurry or whole cell lysate further comprises an additive. In some embodiments, the additive is selected from the group consisting of PEG, maltose, sorbitol, sucrose, glucose, mannitol, lactose, milk powder, starch, and combinations thereof. In some embodiments, the additive is added in an amount of about 0.1 w / v% to about 10 w / v%, such as about 0.5 w / v% to about 5 w / v%, of the cell slurry or whole cell lysate. For example, the additive may be added at 0.5 w / v%, 1.0 w / v%, or 5 w / v% of the cell slurry or whole cell lysate. In a preferred embodiment, the additive is mannitol, sorbitol, sucrose, or a combination thereof.

[0304] In some embodiments, the cell slurry or cell lysate may be lyophilized. For example, the cell slurry or cell lysate may be lyophilized over a period of two days. Methods of lyophilization are known in the art.

[0305] The inventors have found that adding an additive to a cell slurry or whole cell lysate (as described above) increases the enzyme stability compared to a cell slurry or whole cell lysate that does not contain the additive, and that lyophilizing the cell slurry or whole cell lysate (as described above) increases the enzyme stability compared to a non-...

Claims

1. (a) Propylene oxide feed, (b) β-cyclodextrin feed, (c) Mass flow meter or mass flow controller, and (d) Static mixer A hydroxypropyl-β-cyclodextrin (HPBCD) reactor system equipped with the above features.

2. The propylene oxide feed is pressurized and / or The reactor system according to claim 1, wherein the β-cyclodextrin feed is pressurized.

3. The reactor system according to claim 1, comprising at least two propylene oxide feeds, wherein optionally, the at least two propylene oxide feeds are operably connected to a separate mass flow meter or mass flow controller.

4. (i) further comprising a back pressure regulator, wherein the back pressure regulator is optionally operably connected to a plug flow reactor or a coiled tube, (ii) further comprising a mass flow controller and / or (iii) The reactor system according to claim 1, further comprising a temperature controller, wherein the temperature controller optionally maintains a temperature of approximately 30°C to approximately 60°C.

5. The reactor system according to claim 1, wherein the β-cyclodextrin feed contains NaOH, and optionally the β-cyclodextrin feed contains NaOH at a concentration of about 5 to about 10 equivalents.

6. The reactor system according to claim 1, wherein the static mixer is a helical static mixer and / or one or more of the feeds are operably connected to a syringe pump.

7. The reactor further comprises a coiled tube and / or a plug flow reactor, optionally (i) The plug flow reactor comprises at least two coiled tubes and a temperature control unit, wherein optionally at least one supply amount of propylene oxide is supplied in front of the first coiled tube and at least another supply amount of propylene oxide is supplied in front of the second coiled tube, or (ii) The reactor system according to claim 1, wherein at least one amount of propylene oxide is supplied before the plug flow reactor.

8. The reactor system according to claim 1, wherein the propylene oxide is supplied at at least two locations.

9. It is further equipped with collection tanks, and optionally, (i) The collection tank is operably connected to an acid feed, and optionally the acid feed contains hydrochloric acid, sulfuric acid, lactic acid, acetic acid, formic acid, citric acid, oxalic acid, uric acid, malic acid, fumaric acid, tartaric acid, or a combination thereof; or (ii) The reactor system according to claim 1, wherein the system provides a total residence time of approximately 30 minutes to approximately 70 minutes.

10. The reactor system according to claim 1, wherein a first propylene oxide feed provides a concentration of about 7 to about 15 equivalents, and a second propylene oxide feed provides a concentration of about 3.5 to about 15 equivalents.

11. The reactor system according to claim 1, further comprising a purification process including a container for contacting a crude mixture of HPBCD with activated carbon, wherein the purification process optionally further comprises a sterile filter.

12. A method for producing a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, (a) Contacting a hydroxypropyl-β-cyclodextrin (HPBCD) mixture with at least two solvents, wherein the HPBCD mixture includes highly substituted HPBCD and less substituted HPBCD, (b) Dissolving the highly substituted HPBCD in one of the solvents, (c) Removing the low-degree substituted HPBCD by precipitation. The method, including the method described above.

13. A method for producing a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, (a) Contacting a hydroxypropyl-β-cyclodextrin (HPBCD) mixture with at least two solvents, wherein the HPBCD mixture includes highly substituted HPBCD, (b) Dissolve the highly substituted HPBCD in one of the solvents to form a mother liquor. (c) Filtering the mother liquor. The method, including the method described above.

14. The method according to claim 13, further comprising freeze-drying the mother liquor to obtain a solid, and optionally further comprising analyzing the solid by MALDI-TOF to determine the degree of substitution.

15. The method according to claim 13, wherein the at least two solvents include ethanol and acetone.

16. A composition comprising a mixture of methylated 2-hydroxypropyl-β-cyclodextrin (HPBCD) having a degree of substitution of approximately 6.5 to approximately 9.5, and methylated glucose having 0 to 5 2-hydroxypropyl groups.

17. The composition according to claim 16, wherein the composition has a mass spectrum as shown in Figure 25.

18. A method for oligomer substitution of a hydroxypropyl-β-cyclodextrin (HPBCD) mixture by methanolysis, (a) Mix HPBCD and methanol. (b) Stir until the HPBCD is dissolved. (c) Adding an acid to the mixture, (d) Heat the mixture to at least about 50 to about 90°C. (e) Stir the mixture and maintain its heat for at least about 24 hours. (f) Neutralizing the mixture with a base, (g) Filter the mixture. The method, including the method described above.

19. A method for purifying a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, (a) Purify the HPBCD mixture by nanofiltration. (b) Collect nanofiltration permeates equivalent to at least five diafiltration volumes in total, and (c) Freeze-dry the resulting residue to obtain solid hydroxypropyl-β-cyclodextrin. The method, including the method described above.

20. (i) The purification is carried out at a feed pressure of about 200 psi to about 400 psi; (ii) The purification by nanofiltration includes a flat sheet membrane, and optionally the flat sheet membrane is 0.010 to 0.050 m ２ Including the area of; (iii) The method comprises collecting nanofiltration permeates for a total of at least seven diafiltration volumes; and / or (iv) The method according to claim 19, wherein the method comprises collecting nanofiltration permeates for a total of at least 10 diafiltration volumes.

21. A method for purifying a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, (a) Purify the HPBCD mixture by nanofiltration. (b) Collect nanofiltration permeates equivalent to at least five diafiltration volumes in total, and (c) Analyze the propylene glycol content of the remaining residue obtained. The method, including the method described above.

22. (i) further comprising freeze-drying the obtained residue to obtain solid hydroxypropyl-β-cyclodextrin; and / or (ii) The method according to claim 21, further comprising step (d) contacting the HPBCD mixture with activated carbon.

23. A method for producing a hydroxypropyl-β-cyclodextrin (HPBCD) mixture, (a) Contacting the first propylene oxide feed with the beta-cyclodextrin feed to form the first reaction effluent, and (b) Including contacting a second propylene oxide feed with the first reaction effluent to form a second reaction effluent, The method wherein the second reaction effluent comprises a mixture of HPBCD containing an unsubstituted beta-cyclodextrin molecule and a beta-cyclodextrin molecule substituted with a hydroxypropyl group at one or more hydroxyl positions.

24. A composition produced by the method described in any one of claims 12 to 15 or 18 to 23, wherein the composition is (i) comprising a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture contains 0% to 0.3% unsubstituted beta-cyclodextrin ("DS-0") or 0% to 1% beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to patients requiring it; or (ii) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and at least 70% of the beta-cyclodextrin is DS ａ It has a DS within ±1σ, where σ is the standard deviation; or (iii) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 1% to 10% beta-cyclodextrin substituted with seven hydroxypropyl groups ("DS-7"); or (iv) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 25% or less beta-cyclodextrin substituted with four hydroxypropyl groups ("DS-4"); or (v) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 20% or less of beta-cyclodextrin substituted with five hydroxypropyl groups ("DS-5"); or (vi) comprising a mixture of beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture contains 0% to 2.5% of beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the composition is suitable for intrathecal, intravenous, or intraventricular administration to patients in need; or (vii) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture comprises 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the mixture comprises 5% to 25% beta-cyclodextrin substituted with six hydroxypropyl groups ("DS-6"); or (viiii) A mixture comprising beta-cyclodextrin molecules substituted with hydroxypropyl groups at one or more hydroxyl positions, wherein the mixture contains 0% to 1% unsubstituted beta-cyclodextrin ("DS-0") and beta-cyclodextrin substituted with one hydroxypropyl group ("DS-1"), and the following structure: 【Chemistry 1】 It contains a beta-cyclodextrin having a glucose unit, where R １ , R ２ , and R ３ R is independently -H or -HP at each occurrence site, and HP contains one or more hydroxypropyl groups, and is HP in the beta-cyclodextrin. １ and R ２ The composition wherein the percentage of the total number of occurrences of the combinations is in the range of 85% to 95%.

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