JP2004533470A - Fluid microbe control system - Google Patents

Abstract

An arrangement for controlling the content or growth of microorganisms in a fluid, comprising: a) a zeolite having micropores loaded with an agent, wherein the agent has an affinity for the zeolite micropores; Comprising a zeolite having antiseptic properties and b) the fluid, wherein the zeolite and the surrounding fluid are at least partially in contact or arranged to provide at least partial contact. Placement. The fluid contains molecules having a defined affinity for the zeolite that is larger than the zeolite's micropores and lower than the agent. The fluid contains a therapeutically active compound or composition in a therapeutically effective amount and concentration, and the total amount of agent in the zeolite is greater than the amount corresponding to the preservative level for the fluid. . Also disclosed are methods, uses, and zeolites for controlling the content or growth of microorganisms in a fluid.

Description

【００３３】
実施例４
本実施例は、ゼオライトに吸着されたｍ−クレゾールで処理した後のStaphylococcus aureus (ATCC 6538)，CFU ml-1)の増殖阻害を示す。
【００３４】
Eur. Pharm. 2nd Rd. VIII 14 (1992) に従った試験。抗菌性保存剤の効力。脱アルミニウム化ゼオライトＹ（ＤＡＹ，６３〜１２５μｍ粒子）がｍ−クレゾール５ｍｇ／ｍｌとインキュベートされた後、そのゼオライトはそれが吸い取られてからからにされ、更に暖小室中で一晩乾燥された。次いで、０．２ｍｇグリシン／ｍｌ及び４１ｍｇマンニトール／ｍｌを有し、成長ホルモン（ＧＨ）を添加されたか又はされていないリン酸緩衝液（０．３３３６ｍｇ／ｍｌのリン酸一ナトリウム塩、０．７０６４ｍｇ／ｍｌのリン酸二ナトリウム塩）中に、そのゼオライトは懸濁され、その後にそのゼオライトは、Staphylococcus aureus と３７℃でインキュベートされた。
【００３５】
【表２】

【００３６】
実施例５
本実施例は、超安定ゼオライトＹ（ＵＳＹ）への塩化ベンザルコニウムの吸着を示し、それは、図８中の図表を参照して記載される。
【００３７】
塩化ベンザルコニウム（１５６〜２０ｍｇ／ｍｌ）が６０分間ロッカーテーブル上で超安定ゼオライトＹ（ＵＳＹ粒子６３〜１２５μｍ）の２５ｍｇ／ｍｌとインキュベートされた。遊離塩化ベンザルコニウムの量は、２６３ｎｍ吸光度によって測定され、そしてそのゼオライトに吸着された塩化ベンザルコニウムの量が計算された。
実施例６
本実施例は、超安定ゼオライトＹ（ＵＳＹ）からの塩化ベンザルコニウムの放出を示し、それは、図９中の図表を参照して記載される。
【００３８】
超安定ゼオライトＹ（ＵＳＹ粒子６３〜１２５μｍ）が、塩化ベンザルコニウム５ｍｇ／ｍｌとインキュベートされ、その後、そのゼオライトはそれが吸い取られてからからにされ、更に暖小室中で一晩乾燥された。吸着された塩化ベンザルコニウムを有するゼオライトは、最初はＰＢＳ（■）中に、次いで、９５％エタノール（●）中に、繰り返し２０ｍｇ／ｍｌに懸濁された。その溶液中の塩化ベンザルコニウムの濃度が、２６３ｎｍにおける吸光度によって測定された。
実施例７
本実施例は、超安定ゼオライト（ＵＳＹ）からのセファロチンの放出を示し、それは、図１０中の図表を参照して記載される。
【００３９】
超安定ゼオライト（ＵＳＹ）は、防腐剤セファロチン５ｍｇ／ｍｌとインキュベートされ、その後、そのゼオライトはそれが吸い取られてからからにされ、更に暖小室中で乾燥された。吸着されたセファロチンを有するゼオライトは、繰り返し、１０ｍＭグリシンｐＨ２．５液（■）中で２０ｍｇ／ｍｌに懸濁され、そしてその溶液中のセファロチンの濃度は、２６０ｎｍにおける吸光度測定によって測定された。次いで、そのゼオライトは、１０ｍＭリン酸緩衝液ｐＨ８．０（●）中に懸濁された。ｐＨの変化は、セファロチンを脱プロトン化させ、そして導入された電荷はそのゼオライトからのセファロチンの放出の増大をもたらした。
実施例８
本実施例は，種々の抗生物質の吸着及び放出を示す。超安定ゼオライトＹ（ＵＳＹ）は、種々の抗生物質とインキュベートされ、そしてそのゼオライトに吸収された量が計算された。次いで、そのゼオライトは乾燥されて、緩衝液中で２０ｍｇ／ｍｌに懸濁され、そして、その溶液中の放出された抗生物質の濃度が測定された。
【００４０】
【表３】

【００４１】
実施例９
脱アルミニウム化ゼオライト（ＤＡＹ；６３〜１２５ミクロン）が、ｍ−クレレゾール液（１０ｍｇ／ｍｌ）で、４０ｍｇＤＡＹ／ｍ−クレゾール溶液ｍｌであるゼオライト含有量で負荷された。E. Coli (CU1867, ATCC♯47092) 懸濁液が、未負荷ゼオライト及びｍ−クレゾールで負荷されたゼオライト混ぜられて、５分間沈殿できるようにされた。その上澄み液が、ゼオライト沈殿物から除かれて、残存する囲まれた細菌懸濁液（８×１０^６コロニー形成ユニット、ＣＦＵ）を有するゼオライトが、室温でインキュベートされた。１８時間後、その沈殿物は再懸濁されて、ＬＢ−ａｇａｒ上でのコーティングの後にＣＦＵの数が測定された。その試験は、緩衝液（０．３３３６ｍｇ／ｍｌリン酸一ナトリウム塩、０．７０６４ｍｇ／ｍｌリン酸ニナトリウム塩、２ｍｇ／ｍｌグリシン及び４１ｍｇ／ｍｌマンニトール）中でも、成長ホルモン（ＧＨ，５．５ｍｇ／ｍｌ）を含有する緩衝液中でもなされた。２つの試験からの分布は、以下に、ＣＦＵで与えられる。
【００４２】
【表４】

【００４３】
実施例１０
脱アルミニウム化ゼオライト（ＤＡＹ；６３〜１２５ミクロン）が、ｍ−クレゾール（１０ｍｇ／ｍｌ）と、４０ｍｇＤＡＹ／ｍ−クレゾール溶液のゼオライト含有量で負荷された。E. Coli (CU1867, ATCC♯47092) 懸濁液が、未負荷ゼオライト及びｍ−クレゾールで負荷されたゼオライト混ぜられて、その混合液は５分間沈殿できるようにされた。その上澄み液がゼオライト沈殿物から除かれて、残存する囲まれた細菌懸濁液（８×１０^６コロニー形成ユニット、ＣＦＵ）を有するゼオライトが、室温及び＋８℃のそれぞれでインキュベートされた。その試験は、ＬＢ−培地中でなされて、室温で１８時間後及び＋８℃で２．５日後のそれぞれでその沈殿物が再懸濁されて、そしてＬＢ−ａｇａｒ上でのコーティングの後にＣＦＵの数が測定された。その結果は、以下にＣＦＵで与えられる。
【００４４】
【表５】

【図面の簡単な説明】
【００４５】
【図１】
図１は、理想状態のもとでの意図された本発明の平衡系を箱の形態で概略的に示す。
【図２】
図２は、初期に高い飽和度へ負荷されたゼオライトについての溶出曲線を概略的に示す。
【図３】
図３は、ゼオライト中に剤量の勾配が形成されることができるほどの十分な長さであるカラムの形態のゼオライトベッドを概略的に示す。
【図４】
図４は、単一のチャンバーに関連した種々のゼオライト配置を概略的に示す。
【図５】
図５は、上流チャンバー及び下流チャンバーに関連した種々のゼオライト配置を概略的に示す。
【図６】
図６は、実施例１に関連した図表を示す。
【図７】
図７は、実施例２に関連した図表を示す。
【図８】
図８は、実施例５に関連した図表を示す。
【図９】
図９は、実施例６に関連した図表を示す。
【図１０】
図１０は、実施例７に関連した図表を示す。【Technical field】
[0001]
The present invention relates to an arrangement for controlling the content or growth of microorganisms in a fluid. The arrangement comprises a) a zeolite having micropores loaded with an agent, the agent having an affinity for and properties of the zeolite micropores, and b) a fluid. Wherein the zeolite and the fluid are at least partially in contact, or are arranged to provide for at least partial contact. The present invention also relates to suitable zeolites for their arrangement and methods of using them.
[Background Art]
[0002]
The use of preservatives to control microbial content or growth is common for a variety of purposes, such as cleaning and sterilizing fluids and preservative additives to fluids. A common problem is that they tend to be toxic or harmful not only to their microorganisms, but also to other organisms, including humans and animals, and minimizing the amount of agent desirable. On the other hand, they often must be used in excess, corresponding to the worst anticipated microbial exposure for the fluid in its intended use. The problems are exacerbated when the fluid is intended to be exposed to the body, for example, as a respiratory gas for body treatment or as a formulation. This is most applicable for injectable formulations. Although the formulation severely limits the type and amount of preservative, it still has the necessary side effects that must be weighed against the therapeutic value of the formulation treatment.
[0003]
Efforts are being made to better control the amount and exposure pattern of the preservative. One proposal is to absorb the preservative into a carrier to control its release rate. EP 301 717 discloses that a medical tube is loaded with a zeolite loaded with a metal ion such as Ag, Cu or Zn as an antimicrobial agent in order to prevent long-term infection in the body tissue surrounding the inserted tube. It is proposed to coat with. However, the degree of control obtained was only limited. Its emission is defined solely by diffusion and cannot be varied or actively controlled. The zeolite must also be applied in thin layers for efficient use, since the effect extends only to thin layers. The proposed ion exchange system limits the agents absorbed to certain metal ions. The use of zeolites as an absorbent for orally administered medical agents, as exemplified by EP240169, can avoid the occurrence of peak concentrations, but cannot be controlled to obtain low, uniform concentrations. Run into problems.
[0004]
The document WO 97/15391 proposes the use of zeolites to both absorb preservatives from pharmaceutical formulations and expel them, in order to reduce the amount of preservatives delivered to the body. The zeolite may be placed at the forefront of a syringe-type device so that the formulation is displaced and expelled. However, the location of the zeolite between the formulation and the outlet opening for the formulation means that in the most infectious tracts, i.e. in the open outlet openings, a low content Since only this preservative is present, infection of this part means that sufficient preservative is not available to control its growth. The problem is more pronounced in a multiple dose arrangement. This is because the liquid in the dead space in front of the zeolite absorbent has passed through the zeolite and is therefore subject to uncontrolled microbial growth.
[0005]
A similar problem is shown in WO 87/05592 for wastewater treatment. Here, the receptor to be protected is a biological bed used to degrade the waste content in the water. The zeolite bed is inserted into the inflow tube in front of the bed and is able to absorb toxic components that occasionally cluster in the incoming water. Thus, the concentration of the toxic substance is kept low and not so high that the biological bed can deteriorate. Because this system simply absorbs any toxic components in the incoming wastewater, it does not use the toxic components for preservative purposes, or to say nothing more efficiently, and No control is provided for the purpose.
[0006]
Thus, there is still a need for improved methods and means for controlling the content or growth of microorganisms in a fluid.
[Summary of the invention]
[0007]
The main objective of the present invention is to provide a system for controlling the growth of microorganisms in a fluid, which avoids the disadvantages and disadvantages of the technology used previously. A more specific objective is to provide a system based on the controlled and efficient use of preservatives. Another object is to provide a system which allows to maintain the preservative action and reduce the amount of agent or to maintain the amount of agent and improve the preservative action. Yet another object is to provide a system that exposes the fluid to an amount of agent adapted to its current microbial load, reducing the need for an excess amount of agent in the worst case. A further object is to provide a system that provides a preservative barrier between the entrapped fluid and its surrounding environment, independent of the fluid content of the agent. Yet another object is to provide a fluid that is a pharmaceutical formulation with a low agent content that is low enough to allow for exposure to humans and animals, including injection of the treated fluid into the body. The aim is to provide a system that gives a fluid that is also affinity. A further object is to provide a system applicable to both small and large volumes of fluid in static and intermittent or continuous movement or administration. Yet another object is to provide a system that allows for the use of a wide range of preservatives. Yet another object is to provide a system that can be used for both gas and liquid fluids. Yet another object is to provide a system that allows for flexible design of devices and arrangements.
[0008]
These objects are achieved with the characteristics indicated in the appended claims.
In the system of the present invention, zeolites are used to absorb preservatives. In general, zeolites are highly inert and structurally rigid as compared to other sorbents, which can be formed into very flexible forms into structures with variable bulk porosity. Can and is a selective and efficient absorbent due to its high pore volume and uniform pore size. In addition, zeolites can have their pore sizes varied to accommodate preservative target molecules. A further important advantage in the present context is obtained in hydrophobic zeolites, ie zeolites having a high content of silicon and a low content of aluminum in their crystal lattice backbone. These zeolites are more inert and stable and are particularly important when the fluid is exposed to humans or animals. They have no or low tendency to release particles and aluminum ions, which is important, for example, when the fluid is a pharmaceutical formulation. They withstand high temperatures and prolonged exposure to water without degradation, such as the sterilization and long-term storage of pre-loaded devices, or repeated operation in a continuous cleaning sequence. Or it is important to be able to play. Lastly, these hydrophobic zeolites have expanded the range of preservatives that can be used by their affinity for hydrophobic and non-ionic compounds, which means that these classes of compounds can be exposed to humans. Important because it covers a number of preservatives that are acceptable and works particularly well with the principles of the present invention. The present invention relates to a zeolite pre-loaded with a preservative, wherein the fluid in contact with the pre-loaded zeolite achieves the preservative level of the agent by controlled release from the zeolite. Utilizes preloaded zeolite. Accordingly, the present invention provides for the unconventional conventional properties of these zeolites, particularly their ability to absorb an amount of agent such that the fluid in contact with the zeolite increases its content to preservative levels. Use. This is, for example, the application of zeolites to filtration, where the fluid has its agent content reduced from preservative levels to below preservative levels while the agent increases in zeolite filters to medium levels In contrast to applications that do. This new use of zeolites serves several purposes of the present invention. The loaded zeolite acts as a buffer and releases additional agent when needed, e.g., when a new fluid volume is contacted or when the agent is consumed by microorganisms or otherwise. As provided, the level of preservative in the fluid can be kept low and need not be raised to excessive levels for worst case situations. Without being bound by theory, the fluid / zeolite system attempts to establish an equilibrium with a certain level of agent in each of the fluid and the zeolite, and releases the agent by the zeolite regardless of any disturbance. Or it is hypothesized that uptake will lead to a new equilibrium drive. Thus, low controlled consumption and the level of agent in the fluid that is always adequate can be maintained despite the greater amount absorbed by the zeolite, and at least to some extent the quasi-static ) Ensure that equilibrium with fluid concentration is repeatedly established. The fluid and zeolite can be statically contacted with the zeolite mass or powder by passing the fluid over a zeolite surface or coating or through a bed or column of the zeolite. Hence, the system is compatible with static, intermittent and continuous arrangements, and is also compatible with establishing equilibrium by diffusion, mixing or forced flow. In addition, the loaded zeolite has excellent barrier properties in its vicinity, since the zeolite has a really high agent content and microorganisms tend to grow preferentially on surfaces rather than in most of the fluid To prevent infection at its surface, through its stomas and at cracks, defects and dead spaces that may be present. Its barrier properties reduce the need for agents in the highest risk of infection, for example, between the fluid and its opening to its surroundings, as additional precautionary measures, or for large volumes of fluid. As a means to do so, it can be used to place the zeolite. It does not prevent the system of the present invention from being combined into a final filtration step in which the already low agent content in the fluid is further reduced. This can be done to minimize exposure or to recover the agent for destruction or feedback to the zeolite to create a static preservative system that allows for continuous fluid processing. Can be done to The loaded zeolite of the present system allows for reversibly controlled release of the agent, both when the fluid is a gas and when the fluid is a liquid, and the combined flexibility of the system. Make the present invention suitable for many different applications as illustrated.
[0009]
Further objects and advantages of the present invention will become apparent from the detailed description below.
DETAILED DESCRIPTION OF THE INVENTION
[0010]
Definition
As used herein, a "system" is generally described, claimed, exemplified or implemented as one or more devices / arrangements, methods, uses or combinations thereof. Is understood to refer to the principles of the present invention.
[0011]
Unless expressly stated or expressly stated to the contrary, expressions such as "comprising," "including," "having," "with," and the like, as used herein, Terms similar to are not understood to be exclusively limited to the listed device elements, compositional compounds / components, or method steps, but are to be understood as tolerating the presence of additional elements, compounds / components and steps. You. It is understood that it covers any device element in whole, subdivided or aggregated form, and that it is "connected," "attached," "placed," "adapted," The phrase "between" and similar terms are not to be understood as exclusively covering the direct contact between the listed elements, but one or more elements or structures intervening there between. Is understood to allow the presence of When used in describing force or action, the same applies for similar expressions. Similarly, unless expressly stated or expressly stated to the contrary, such a representation is a composition that is in the form of any physical or chemical aggregate or mixture with the compound / ingredient interposed therebetween, where possible. It is understood that the state of the compound / component, or assembly, as well as the method steps in any time sequence.
[0012]
Again, "microorganisms" in the present context are understood to mean any organism that can survive as a single cell in the presence of nutrients or host organisms, for example, protozoa such as amoeba. More precisely, the concept has the usual meaning of "microorganism", ie bacteria and fungi of the mold or yeast type.
[0013]
For the purposes of the present invention, expressions such as "preservative condition", "preservative concentration", "preservative level", "preservatively effective" and similar terms in the present context refer to at least one living thing. If and when, the microorganism is in contact with any component capable of serving as a nutrient for the microorganism, at least to slow its growth, preferably to stop its growth, and most preferably to kill it. It is understood to refer to adapted or suitable conditions, but the expressions are not to be understood as requiring the presence of such nutrients. These expressions are also understood to include precautionary situations that do not require the presence of microorganisms. However, the expressions are understood to exclude situations and conditions whose contact or remaining time is not sufficient for any significant action between the preservative and the microorganism. A system that has sufficient preservative action against a single microorganism may require additional agents when the microorganism absorbs or interacts with the preservative, given the possibility according to the invention of using the loaded zeolite as a buffer. Is understood to release and restore the agent content of the fluid, and excludes the option that the fluid has an amount or concentration sufficient to be active against more than one or many microorganisms is not. Indeed, the required dosage can vary for different dosage types. For example, low for antibiotics with well-defined metabolism, high for bold oxidizing or toxic agents. Thus, for example, the conditions usually termed bactericidal and bacteriostatic are included in the meaning of these expressions.
Zeolite
In general, zeolites can be described as a crystalline framework of aluminum silicate or tectosilicate, which can be characterized by their compounds and crystal structure. Generally, the compound is described by a Si / Al ratio. High silica zeolites have less skeletal charge and are commonly referred to as hydrophobic. The opposite is true for high alumina zeolites classified as hydrophilic. Zeolites suitable for this purpose have the general structural formula (AlO₂)_x(SiO₂)_yAnd the ratio y / x may be described as having a general structural formula that may have different values as described below. In this zeolite framework, other ions, such as P, B, Fe, Ga, etc., are substituted for Al and Si to a certain extent and can also be used for the purposes of the present invention. The zeolite framework has a cation attached to each Al atom or other atom having a maximum valency less than 4, and an anion attached to each atom having a maximum valency greater than 4. Zeolites may contain more or less water.
[0014]
The crystal lattice provides a pore system in which the pores are highly uniform in size, but the size is somewhat different between different zeolite types. Generally, the pores comprise a main cavity and an outlet opening. The size of the main cavity varies between different zeolites with a diameter of about 3-11 °, and its outlet is about 1-3 ° smaller. These uniform pores are responsible for the high and selective absorption of the zeolite and are referred to herein as micropores. The zeolite crystals also have microcracks caused by their chemical and physical manufacturing and processing history, they are not uniform and, for this purpose, have low cracking It is preferred to use zeolites with or without zeolite. Finally, zeolite crystals are usually of limited size and form a powder or particle mass. Such a compact may itself be used, for example, to be suspended in a fluid for absorption. By sintering or by adding adhesive components, such as bentonite, talc or phosphate glass, the compact can be consolidated into any shape and size configuration. In doing so, macropores may be left between the particles. For both the unconsolidated powder and the compacted form, the macropores between the crystal grains are called bulk porosity, and the total bulk capacity of the zeolite compact or form, respectively. Expressed as the total macropore volume for Bulk porosity serves the purpose of allowing the fluid to enter the macropores and contact the individual particles, and to allow the fluid to pass through the zeolite compact or shape. . Suitable bulk porosity for such purposes can be at least 20%, preferably at least 30%, and most preferably at least 35%, and for stability reasons, among other things, The porosity can be at most 90%, preferably at most 80%, and most preferably at most 75%. Indeed, known means other than pore porosity to create permeability and / or to increase the contact surface between the fluid and the zeolite, such as narrow channels through a zeolite bed or body, zeolites Can be used in other ways or in combination.
[0015]
Zeolites having a y / x ratio of 15 or less for this purpose are said to be hydrophilic, while zeolites having a y / x ratio higher than 15 are said to be hydrophobic. Either type can be used for the purposes of the present invention. Hydrophilic zeolites can be used, for example, when preservatives are also hydrophilic. If the agent is ionic, it is also possible to take advantage of the well-known possibility of impairing its affinity for zeolites by altering the surrounding ionic properties, such as pH, and Reduces its affinity in environments that become non-ionic. This can be important to fine tune the release of the agent and create a self-regulating system or to make that same loaded zeolite useful for different fluid requirements. Hydrophilic zeolites can have a y / x non-rate less than 15, for example less than 5, and even less than 1. For the reasons indicated, it is often preferred to use dealuminated zeolites or, for example, hydrophobic zeolites also referred to as ultrastable zeolites due to their high stability, and also to use a large class of hydrophobic preservatives and nonionics. May be used, the latter providing stable affinity properties regardless of the ionic properties of the fluid. The hydrophobic zeolites preferably have a y / x ratio higher than 100, more preferably higher than 200, and most preferably higher than 1000. Suitable zeolite types can be, for example, silicalite, mordenite and especially zeolite Y. With regard to the size of the micropores, the zeolite Y and mordenite types belong to the largest ones today with approximately 7% and 7.5% respectively, whereas silicalite has approximately 5.5% different two-pore sizes. It has the size. Silicalite and zeolite Y have a three-dimensional pore system, whereas mordenite has a two-dimensional pore system and is therefore somewhat less accessible. In a known manner, the hydrophobic zeolite can be obtained through direct synthesis (eg, silicalite) or post-synthesis operations (eg, mordenite, zeolite Y), eg, alkali of zeolite Y, eg, ammonia, with acid, For example, it can be produced, for example, by alternating treatment with hydrochloric acid (USY) or by treatment of zeolite Y with silicon tetrachloride (DAY). Of these post-synthesis engineered zeolites, the alkali / acid treated has been found to be faster in terms of its adsorption and release, but has a high weight (high- weight) has a high tendency to adsorb for proteins, while the opposite is found for silicon tetrachloride treated zeolites.
[0016]
The zeolite may be used as such, or may be treated to modify its properties. It can be, for example, coated with, for example, dextran or polyethylene glycol, to reduce the risk of clogging the microporous system with larger molecules contained in the fluid.
Preservative
In general, preservatives useful for this purpose should have a size suitable to be contained in a zeolite microporous system. Atoms and atomic ions such as Ag or Cu can be used because they are easily accommodated. Preferably, the agent comprises molecules, of which a truly broad range of suitable agent compounds are available. Such molecules should then be able to be accommodated in their micropores, at least partially in the case of elongated or branched molecules, but preferably the entire molecule is accommodated Should. This places restrictions on the size of suitable molecules, and roughly they have a molecular weight of less than 3000, preferably less than 2000, and most preferably less than 1500. Compounds in these ranges that can be accommodated in those micropores are termed "low-weight," while larger molecules that cannot be accommodated are termed "high-weight." Called a molecule. Compounds and molecules are understood to include aggregates, chelates, etc., when they are sufficiently stable to behave as a single entity to the microorganism. A second general requirement is that the agent has sufficient affinity for at least one zeolite type and is sufficient to provide a preservative in a small volume of fluid for at least the zeolite in contact with the zeolite. To be able to absorb as much as possible. As indicated, generally, the choice of agent for the zeolite type controls this.
[0017]
Any low weight preservative that meets the above requirements can be used according to the present invention. Purely oxidizing substances such as chlorine, hydrogen peroxide and the like can be used as well as purely toxic substances such as cyanide, but it is preferable to use an agent having a more selective action. A suitable class of agents are antibiotics that can inhibit growth or kill microorganisms at low concentration levels, such as clindamycin or penicillin. Preferred are those that are non-toxic or only moderately toxic to animals or humans and can be used in medicine. Antibiotics belonging to different groups with respect to their biological mechanisms may be used, such as those that impair cell wall synthesis, protein synthesis, folate metabolism, nucleic acid synthesis, and the like. Another suitable class of agents are those commonly referred to as preservatives, for example, halogenated compounds such as DDT, triclosan, or the like, or aromates or polyaromates. For medical applications, it is preferable to use preservatives approved for such use. Suitable compounds of this type include benzyl alcohol, benzalkonium chloride, cetrimide, cholbutol, chlorohexidine, chlorocresol, hydroxybenzoates, phenyl alcohol, phenoxy alcohol, phenylmercuric nitrite, chloramphenicol Etc. are included. Good results have been obtained with phenol and cresol, especially m-cresol.
[0018]
The above groups and classes of agents have been made for ease of reference only and are not to be considered limiting as long as the functional requirements are met. It is well possible to use mixtures and combinations of agents within or between the classes and within or between the hydrophilic and hydrophobic types.
Load zeolite
The zeolite may be loaded by any known method, for example by contacting it with an agent such as a mixture, suspension, emulsion or the like of the medium in pure form or in liquid or gaseous form. The contacting operation can be performed, for example, by bringing the agent into contact with the zeolite, being stirred together, or by passing the agent over or through the zeolite, And the latter allows a gradient of agent to be created in the zeolite. The contacting can be performed at different temperatures, for example, at elevated temperatures to accelerate the operation. The zeolite may be subjected to various post-treatments, such as sterilization operations based on irradiation, chemical treatment, heating, etc., a drying step to provide a stable semi-finished product for storage, pre-loading the device before contact with the fluid, or Is subjected to an additive treatment step for modifying the properties of All these operations are facilitated by the stability properties of the zeolites. Some loss of agent can occur, for example, during heating or vacuum treatment, which should be disabled by adding a replenishing amount of agent before or after such a step. For example, a syringe pre-filled with the formulation may require a sterilization step, and if a zeolite is present, it will be affected by the same treatment. Similarly, some formulations may be subjected to a freeze-drying or freeze-drying step under vacuum, and any zeolites present there may be subjected to the same treatment. During such steps, some agent is lost from the loaded zeolite, but can be replenished by a corresponding initial overload. Especially at high loads, some agents are absorbed more slowly than most agents, possibly due to some absorption outside the micropores, for example, on the outer surface or in microcracks. But, again, it is sometimes observed. Mismatches introduced by such factors include, for example, subjecting the loaded zeolite to a short wash or elution step to remove loosely bound agents, or removing unloaded or less loaded zeolites. By being inserted downstream of the loaded zeolite, the first released agent is trapped in its downstream micropores until it has the same degree of saturation as its primary loaded zeolite, which is avoided. Is preferred.
[0019]
The minimum requirement for agent and zeolite in the loaded zeolite is that the small volume of fluid to which the loaded zeolite is directed reaches the minimum preservative level of the agent in the fluid when contacting the loaded zeolite. The substantial concentration or distribution of the agent between the fluid and the zeolite is reached. These minimal conditions can be useful, for example, when in contact with air or when the moistened loaded zeolite is no longer used as a preservative barrier. Generally, it is preferable to have more than the minimum conditions. For example, an agent for administering a formulation to one or more, eg, a preservative level in a larger volume of fluid, eg, a predetermined volume surrounding or passing through the zeolite. Preferably, to provide for the presence of an amount. Higher amounts of these agents are provided, for example, by increasing the volume of the zeolite having a lower concentration of the agent, preferably to increase the concentration of the agent in the zeolite, to minimize the amount of zeolite required. obtain. For very large or unlimited volumes of fluid, a new agent may be added directly to the zeolite or to the fluid to be contacted with the zeolite, or the agent may be separated from the already treated fluid and separated from the zeolite. It is preferred to maintain a minimum or buffer level in the zeolite by continuously or batch replenishing the agent into the zeolite, either directly or by returning to the fluid to be contacted with the zeolite. Increasing the amount of agent in the zeolite above the minimum requirement to ensure that the buffer is present in the zeolite at a level sufficient to provide a safe margin for minimal microbial exposure. It is also preferred that the level be assured, preferably at a level sufficient for worst case exposure for the intended use. As indicated, this can be accomplished by the present invention without sacrificing any significant increase in the concentration of the agent in the fluid. This is because the equilibrium between the zeolite and the fluid allows the preservative level in the fluid to be restored to the same agent concentration repeatedly and with consumption control. For best performance it is preferred that the agent has a really high affinity for the selected zeolite type. The affinity is herein referred to as the maximum saturation, i.e., the amount of agent in the zeolite at the saturation obtained after a sufficient contact time for stabilization for the zeolite in contact with the pure agent, Expressed as weight percent agent rate. The maximum degree of saturation so expressed can be at least 10%, preferably at least 25%, and most preferably at least 50%. Its maximum degree of saturation is only considered a measure, not necessarily the degree to which the zeolite is loaded for use. When the zeolite is loaded at high saturation and there is not enough column effect to be evaluated at the time of contact, it releases a much higher amount in the beginning than when the later saturation decreases. The decrease in concentration in the fluid on continuous or repeated contact with the fluid is very rapid and approximates an inverse linear function, polynomial or exponential function. A high degree of saturation can be used when this density pattern is desired, for example, to maintain a lower density after a one-time high clean. However, for many purposes, the concentration of the agent in the substantially steady fluid after contact with the loaded zeolite, for example, during steady fluid contact or repeated administration of a larger fluid volume, is reduced. It is desirable to have. To achieve this, the agent loaded on the zeolite should be below the maximum saturation, preferably so low that its drop curve reaches an approximately steady mode. As an indication of such saturation, it is stated that the amount of agent in the zeolite should be less than 50%, preferably less than 30%, and most preferably less than 10% of the amount corresponding to the maximum saturation. For high affinity agent and zeolite combinations, the amount is much lower. Generally, this ratio of actual dosage to dosage corresponding to maximum saturation will be higher than 0.01%, preferably higher than 0.1%, and more preferably higher than 1%. These conditions are less stringent when a column effect is present upon contact. If the loaded column is long enough to provide a substantial equilibrium agent concentration in the fluid after a small partial passage through the column, the amount of agent in the fluid or zeolite will increase during passage through the remainder of the column. No change has occurred, and the condition at the exit will be highly steady. A parameter that is relevant for this purpose is the concentration of the agent in the fluid in equilibrium with the loaded zeolite. This concentration can be lower than when not utilizing the principles of the present invention. For example, less than 0.25 times such a concentration, preferably less than 0.1 times the concentration expected in a given application, and most preferably less than 0.05 times. Giving absolute concentration values is difficult, for example, because of the different preservative efficacy of the different classes of agents, and the differences between technical and therapeutic applications. A typical demanding application is a solution for body injection, where the typical agency approved concentration of the preservative is about 2-3 mg / mL, and the general reduction above is It is possible. Even if the value drops to 0.01 mg / mL, it is still proven to be effective. Similarly, it is difficult to give an absolute loading value, but as a guide, the charging rations w / w of the agent on the zeolite is generally higher than 0.1%, preferably higher than 1%. And most preferably greater than 5%, even 10% or 20%.
[0020]
Given the possible changes, the above given considerations are considered primarily as guidelines. If the contact time between the zeolite and the fluid is too short to produce equilibrium, or if the combination of the agent and the zeolite is slowly going to equilibrium, adaptation to higher agent volumes may be required. Temperature should also be considered. Generally, the agent equilibrium concentration in a fluid in contact with a given loaded zeolite increases with temperature. Microorganisms can also be more active at higher temperatures. In most cases, the working temperature is dictated by the constraints of use, but the fluid / zeolite combination may be somewhat self-regulating.
fluid
The system can be used to control microbial growth in a wide range of fluids. The fluid may be a pure substance or a mixture of components that are aggregates in the same or different states. The gas as a continuous phase may contain droplets or solid particles. The liquid as the continuous phase may contain solid particles or liquid droplets, and the mixture may be a suspension, emulsion or the like. The fluid may be a simple mixture such as air or an aqueous solution or mixture, or may be a complex mixture that can even contain unknown contents, for example, contaminated running water or bodily fluids. Preferably, the system is used for fluids that do not interfere too strongly with the described release mechanism. Compounds or compositions that can precipitate or adhere to the zeolites to the extent that they close the micropores or macropores should be contained in the fluid in only small amounts. Preferably, the fluid has a viscosity that is not too high, eg, less than 10,000 cP, preferably less than 1000 cP and most preferably less than 100 cP. The fluid contains a compound that competes with the agent for zeolite affinity, for example, creating a control means for releasing an amount of the agent that varies depending on the abundance or addition of such competitor compounds. obtain. Moreover, the competitor compound is similar to or even greater than the agent, eg, more than 2 times, preferably more than 10 times, and most preferably more than 20 times the affinity of the agent. , Have a defined affinity for zeolites. Such compounds reduce their ionic characteristics, e.g., for hydrophilic and hydrophobic zeolites, respectively, such that the typical elution medium, e.g., the absorbed agent, becomes less affinity for the zeolite. It is also understood to include a medium that acts by increasing it. However, for the reasons outlined, it is often preferred that controlled release of the agent means a substantially steady level of the agent in the fluid. For such purpose, the liquid may be a compound that competes with the agent for affinity with the zeolite, i.e., a compound that is also a low molecule in the sense described above but also has a high affinity for the zeolite. It is preferred to have only a small amount. The fluid may contain large amounts of low molecular compounds. However, they have a lower affinity for zeolite than the agent, for example, at most 0.5 times, preferably at most 0.1 times, and most preferably at most 0.05 times the agent. With different affinities. Such compounds may be air or water molecules with low affinity for hydrophobic zeolites, or small nonionic compounds with low affinity for hydrophilic zeolites. Similarly, the fluid may also contain large amounts of high molecular compounds as defined. Because they do not tend to be absorbed by the zeolites, regardless of their hydrophobic or hydrophilic properties, respectively. Often, in connection with fluids containing such compounds, for example, by using the present invention for medical formulations where the compounds can be proteins, polypeptides, carbohydrate compounds, nucleic acid sequences, etc., these types of It is preferable to use the zeolite characteristic of not absorbing large molecules. As an approximate indicator of "mass" in the above sense, it may be above 0.01 mg / mL, preferably above 0.1, and most preferably above 1 mg / ml. Similarly, "small" can refer to less than these values. The advantages of the present invention are evident, especially when the fluid serves as a microbial nutrient, and especially when such nutrients are present in large amounts. Further, when using the present invention, it is not necessary to include a preservative in the fluid, and therefore it is preferred that the fluid contains no or preferably no other preservative, or only a small amount.
Fluid / zeolite contact
As shown, the fluid and the zeolite are brought into contact in different ways. The contact is effected by holding the fluid static with respect to the zeolite, and typically achieves saturation throughout the fluid volume, relying on diffusion of the agent from the zeolite into the fluid. Prior to being subjected to at least a concentration gradient in the fluid. Contact is also effected by relative movement between the fluid and the zeolite, for example, by stirring the fluid or forcing it to flow through or through the zeolite, typically the fluid. It is possible that there will not be much concentration gradient in it. The zeolite can take the form of particles suspended in a fluid, typically providing a small agent concentration gradient in the zeolite. The zeolite is present in the form of a coating on a surface in contact with the fluid or on a bed or column over which the fluid is moved or through, and typically in the zeolite. Moving from upstream to downstream of the zeolite can result in a concentration gradient for the agent that increases in volume. Beds or columns also have the advantage of promoting contact with the entire fluid, provide an additional barrier effect, and provide bulk porosity or voids that the fluid occupies and improves contact. Columns of inconsequential length also maintain a highly constant agent concentration at the column outlet by saturating the fluid already present at the outlet or middle of the column, and the agent in the zeolite at that outlet It serves the purpose of maintaining the amount substantially constant. For all arrangements, typically the agent concentration in the fluid will increase at least initially upon contact with the zeolite (provided that equilibrium has been achieved by earlier contact or preloading). There is no.). It is advantageous to reach equilibrium or near equilibrium, which can be obtained, for example, when the fluid is left in contact with the loaded zeolite for a sufficient time. However, it is not always necessary to reach such an equilibrium, for example in a fluid flowing system, in order to avoid too long contact times, for example in relation to the speed of the agent / zeolite combination, but it is acceptable. It may be sufficient to reach different preservative level concentrations. Nevertheless, it is possible and preferred to obtain steady state conditions at a suitable preservative level. Generally, the contact occurs by batch operation, intermittent operation or continuous contact. In a batch or intermittent operation, at least for smaller fluid volumes, many, preferably most, and most preferably all of the fluids are contained in the macropores, i.e., the bed has its fluid capacity. There are some advantages to using a zeolite bed or column that has sufficient bulk porosity to contain. This optimizes, among other things, the state of contact during the available remaining time. For intermittent work, it is preferred that the contained fluid volume at least corresponds to the volume of the repeated dose or, in the case of variable doses, to the largest dose considered .
Arrangement
As described, the loaded zeolite has utility itself, e.g., due to its excellent barrier properties, and is just a barrier, e.g., a part that is formed or inserted or sealed in a seal, For example, it may be adapted for a specific application intended to be used as a barrier designed as a patch for attachment over a wound. A preferred arrangement has at least a chamber for the fluid in combination with the zeolite. The chamber may be open, e.g., an open container, or closed, such as a vial or other encapsulation, or in a channel for transporting fluid to the zeolite. It may be part of such a conduit. In such an arrangement, the loaded zeolite helps to keep the contents sterile, e.g., fluid passes out of the chamber over the zeolite, preferably through it and out of the chamber At times, it may act as a barrier to prevent contamination across the opening or to increase the concentration of agent in the fluid. Another preferred arrangement is a first upstream chamber, a second downstream chamber and a loaded zeolite, wherein the fluid content of the agent increases at least as the fluid goes through the upstream chamber to the downstream chamber. One that is capable of being brought into contact with the zeolite in a manner. Such an arrangement may be, for example, a growth control of a fluid transport channel for any purpose, or may be part of a fluid delivery arrangement. For a given reason, it is preferred to apply the zeolite in the form of a bed or a column which allows fluid to pass through the bed.
[0021]
Although the arrangements are intended to interact with the fluid, the arrangement is considered to be part of the system when it is useful or adapted for the purposes of the present invention. The arrangement is a precautionary measure when the fluid does not necessarily come into contact with the zeolite, for example, when the zeolite is to be activated, for example, if the seal is accidentally broken or fails. It can also be useful when used as In some cases, such a limitation may be such that the fluid contact is created at a controlled moment, e.g., at a controlled moment in connection with activation or opening of the pre-filled device. A membrane that is easy to rupture, easy to penetrate or remove, or other seal is inserted artificially when inserted into the arrangement.
[0022]
The general arrangement outlined can be adapted for many applications, and the specific application can affect the details of the arrangement. In addition to their use, further applications already indicated will be illustrated in connection with the figures.
[0023]
Description of the drawings
FIG. 1 schematically shows the intended equilibrium system of the invention under ideal conditions in the form of a box. The box on the left shows the loaded zeolite 1, the box in the middle shows the fluid 2 in contact with the zeolite 1, and the box on the right shows the microorganisms 3 present in the fluid. The zeolite and the fluid are in reversible equilibrium with each other, indicated by arrows 4 and 5. Similarly, the microorganism and the fluid are in reversible equilibrium, indicated by arrows 6 and 7. Arrow 4 indicates, for example, that the fluid is initially under-balanced with the agent, or that the fluid becomes unsaturated, for example, in connection with consumption of the agent by microorganisms or addition of new fluid. Figure 2 shows the flow of agent from the zeolite to a fluid in all situations. Arrow 5 indicates the opposite flow of the agent from the fluid to the zeolite when or when the fluid is supersaturated with the agent, e.g., when the liquid fluid is evaporated or microorganisms are destroyed to release the agent. Show. In most uses, the backflow of the agent according to arrow 5 is less important than the flow as arrow 4. Arrow 6 indicates the flow of the agent from the fluid to the microorganisms that are assumed to be present here. Consumption of the agent according to arrow 6 may reduce the agent concentration in fluid 2, resulting in a recovery of agent flow from zeolite 1 to fluid 2 according to arrow 4. Arrow 7 indicates the theoretical possibility that the agent is released from the microorganisms into the fluid 2, for example in connection with their death or destruction, which is followed by an equalization back to the zeolite by the corresponding arrow 5 Can bring flow. The flow of arrow 7 is highly dependent on the mechanism between the agent and the microorganism and is not always present. It is also absent if dead microorganisms are removed from the system. Box 3 contains any other components or disturbances than microorganisms that act to destroy or consume the agent from fluid 2, such as impurities in the fluid or components in the complex fluid formulation to which the agent can bind, absorb, etc. It should be noted that the following are also indicated. Thus, it is clear that the system shown is highly resilient to any form of disturbance or any form of drug consumption. For the purposes of the present invention, the concentration of the agent in the fluid 2 is maintained at preservative levels but very low so that the fluid is much less harmful than if the buffer capacity of the loaded zeolite were not present. It is desirable to do so. The speed of the system towards a new equilibrium during a disturbance can vary. In general, the situation can be described in which the equilibrium between the zeolite and the fluid is fairly fast at their interface, but slower if diffusion is required due to the volume of fluid not in contact with the zeolite. However, agitation may be used to correct the diffusion delay, or fluid flow may be used. The equilibrium between the fluid and the microorganisms can be slower, but highly dependent on the preservative mechanism. However, this is typically less important for performance through the system.
[0024]
FIG. 2 schematically shows an elution curve 20 for a zeolite initially loaded at high saturation. The vertical axis 21 represents the concentration of the agent in the fluid in contact with the zeolite, and the horizontal axis 22 represents the volume of the fluid or the number of resuspension. The black curve 23 may be described as representing the pattern obtained when the fluid was continuously contacted with the zeolite, and the discrete value 24 may be described as representing the concentration achieved with repeated batch contact. . The curve shows that initially a high concentration 25 is obtained in the fluid, and that concentration quickly drops to a state where more volume or further resuspension gives a substantially constant concentration of the agent in the fluid. Is shown. Dashed lines 26 and 27 indicate, respectively, roughly the upper and lower loadings that can be selected to provide a substantially constant agent concentration in the treated fluid. A suitable initial zeolite loading is preferably by pre-loading the zeolite to a level corresponding to line 26 by eluting or evaporating the agent until the level of line 26 is reached after overloading, e.g., a diluted load. It can be obtained by using a fluid. It should be noted that the curve represents the agent concentration in the fluid but not in the zeolite. If the agent has a high affinity for the zeolite, the zeolite can comprise a vast amount of agent, despite the fact that the concentration of the agent in the fluid is low, and the exiting fluid It allows for repetition of large volumes of fluids or many contacts, with a substantially constant agent concentration therein. Furthermore, it should be noted that the curves shown in FIG. 2 are typical for contact patterns where sufficient dosage variation or gradient does not occur in the zeolite, for example, without column effect. is there. Such a state of equalization may occur, for example, when the zeolite is optionally mixed or agitated with a fluid, or when the bed of zeolite is thin or shallow.
[0025]
Figure 3 shows a zeolite bed in the form of a column 31, generally 30, having a column form of the zeolite bed long enough to allow a dosage gradient to form in the zeolite. FIG. If the fluid remaining time is sufficient for the speed of the equilibrium system, the fluid is typically saturated with an agent while passing through the column height of the early fractionation, after which the fluid is Without exchanging the agent between the fluid and the fluid, ie, both the fluid and the zeolite pass through the remainder of the column unchanged. This allows for a highly steady fluid outlet concentration for large volumes of fluid until the column is depleted of agent so that it no longer provides the target concentration through the entire column. In that figure, column 31 has an inlet end 32 and an outlet end 33. Initially, prior to contact with any fluid, the column is assumed to have a constant agent load throughout its length. Fluid is then passed from the inlet 32 end of the column as indicated by arrow 34 to the outlet end 33 as indicated by arrow 35. After a period of work, the state shown is reached. Typically, a dynamic borderline, indicated by dashed line 36, is formed and the lower portion 37 of the column with unchanged drug volume is removed from the upper portion 38 of the column with reduced drug volume. To separate. The fluid reaches a saturated concentration of the agent while passing through the upper portion 38 of the column between the inlet 32 and the boundary line 36, and this portion of the column forms a gradient of increasing agent volume from the inlet to the boundary. Having. During the passage of the remainder of the column from the boundary 36 to the outlet 33 and the lower part 37, little change occurs in the fluid or in the column, and the column has substantially its initial agent content. No gradient is formed and the fluid has a steady outlet concentration. As fluid continues to be pumped into the column, boundary line 36 moves slowly down until it reaches outlet end 33. Here, the entire column has a gradient in which the amount of agent decreases towards the outlet end, and the concentration in the fluid begins to drop. In contrast to the situation described in connection with FIG. 2, it is no longer necessary to limit the zeolite saturation from the beginning to obtain a stable outlet concentration. Instead, the column effect ensures a steady outlet concentration independent of the initial saturation. Even if a high or maximum degree of saturation could be used, then if the outlet concentration was targeted to be similar to that of the system of FIG. Agents combined with strong agents may be selected.
[0026]
FIG. 4 schematically illustrates various zeolite arrangements, generally designated 40 and here comprising a liquid fluid 42, together with a single chamber 41. The chamber may be closed or may be part of a transport channel or larger system as indicated by dashed line 43. The chamber is also shown with an opening 44, which is in the form of a loaded zeolite in the form of a fixed disk 45, for example in the form of a particulate zeolite arranged between retaining sieves, preferably calcined. It is sealed in the form of a self-supporting body. The loaded zeolite in the opening 44 is now exposed to the surroundings and acts as a preservative barrier against microbial infection of the chamber. At the location shown in the figure, the liquid 42 is not in contact with the zeolite disk 45, the zeolite is considered to be a safety measure, and the liquid does not necessarily have to have a preservative level of the agent. Nevertheless, the openings allow access to the liquid, for example, to be poured or pushed through the porous disc. Also shown is another fixed arrangement of zeolite as a coating 46 on the interior surface of the chamber. Due to the possibility of loading the zeolite with high amounts of agent to give low concentrations in the fluid, it is often possible to use fairly small amounts of the loaded zeolite, and a partial coating may be sufficient. Also shown is a non-fixed amount of zeolite 47 in particulate or powder form, which can be present in the chamber or can be agitated and suspended. The illustrated arrangements protect chambers for various purposes or sizes, for example, enclosures for fluids into a large manufacturing plant, for example, in connection with keeping the plant closed and stored. Can be used to
[0027]
FIG. 5 shows various zeolite arrangements, generally indicated as 50, which are fed into the upstream chamber as indicated by arrow 53 and the upstream chamber for flowing fluid extracted from the downstream chamber as indicated by arrow 54. Shown schematically with 51 and downstream chamber 52. Between those chambers, the loaded zeolite is placed as a coating 55 on the wall, preferably beside the bed through which the fluid is passed. Also described schematically is that the zeolite 57 in particulate or powder form is suspended in the incoming fluid stream 53, for example, through a zeolite feed tube 58, and further suspended downstream, for example, of possible removal. For example, it is possible to collect through a zeolite extraction tube 59, or for its repeated feeding into a zeolite feed tube 58. Such an arrangement may be such that the loaded zeolite pumped into the tube 58 is varied externally to optimize its condition, for example, to minimize the amount of zeolite or to fluctuate in the incoming fluid 53. It can be treated by reloading the agent to adapt the zeolite dosage or agent type, thus providing an additional degree of control. Again, the arrangement shown in this figure can also be adapted for many applications with flowing fluids and for the size of the arrangement, for example, from a small syringe to a large manufacturing or fluid cleaning plant.
【Example】
[0028]
Example 1
This example shows the release of m-cresol from a zeolite with adsorbed m-cresol, which is described with reference to the chart in FIG.
[0029]
After incubation with varying concentrations of m-cresol, the zeolite was allowed to settle and the supernatant was removed. The zeolite is then suspended in PBS (phosphate buffered salin) such that an amount corresponding to 20 mg / ml of dried zeolite is suspended and incubated on a rocker table for 30-60 minutes After that, the concentration of cresol in the supernatant was measured by absorption at 276 nm. Thereafter, the zeolite was suspended in fresh PBS. Cresol concentration was measured again in the supernatant and the zeolite was resuspended in PBS.
Example 2
This example illustrates the release of m-cresol from a zeolite containing adsorbed m-cresol, which is described with reference to the chart in FIG.
[0030]
After incubation with 23.1 mM m-cresol, the zeolite was dried on a glass filter and the zeolite was repeatedly suspended in PBS in an amount corresponding to 5-20 mg / ml of dried zeolite. Between each suspension, the zeolite was incubated on a rocker table. The concentration of cresol in the supernatant was analyzed by absorbance at 276 nm.
Example 3
This example demonstrates the growth inhibition of Staphylococcus aureus (ATCC 6538, CFU ml-1) after incubation with m-cresol adsorbed on zeolite. Test according to Eur. Pharm. 2nd Ed. VIII 14 (1992). Antimicrobial preservative efficacy.
[0031]
Ultrastable zeolite Y (USY, 63-125 μm particles) is incubated with m-cresol 5 mg / ml, after which the zeolite is allowed to drain before it is dried and further dried overnight in a closet. Was done. The zeolite with adsorbed m-cresol was then suspended in PBS and incubated at 37 ° C with Staphylococcus aureus. The concentration of m-cresol in the solution after suspending the zeolite was 0.14 mg / ml.
[0032]
[Table 1]

[0033]
Example 4
This example shows the growth inhibition of Staphylococcus aureus (ATCC 6538), CFU ml-1) after treatment with m-cresol adsorbed on zeolite.
[0034]
Test according to Eur. Pharm. 2nd Rd. VIII 14 (1992). Antimicrobial preservative efficacy. After dealuminated zeolite Y (DAY, 63-125 μm particles) was incubated with 5 mg / ml m-cresol, the zeolite was allowed to drain after it was blotted and then dried overnight in a warm room. Then a phosphate buffer with 0.2 mg glycine / ml and 41 mg mannitol / ml, with or without growth hormone (GH) (0.3336 mg / ml monosodium phosphate, 0.7064 mg / Ml disodium phosphate), the zeolite was then incubated with Staphylococcus aureus at 37 ° C.
[0035]
[Table 2]

[0036]
Example 5
This example shows the adsorption of benzalkonium chloride on ultrastable zeolite Y (USY), which is described with reference to the chart in FIG.
[0037]
Benzalkonium chloride (156-20 mg / ml) was incubated with 25 mg / ml of ultrastable zeolite Y (USY particles 63-125 μm) on a rocker table for 60 minutes. The amount of free benzalkonium chloride was measured by absorbance at 263 nm and the amount of benzalkonium chloride adsorbed on the zeolite was calculated.
Example 6
This example illustrates the release of benzalkonium chloride from ultrastable zeolite Y (USY), which is described with reference to the chart in FIG.
[0038]
Ultrastable zeolite Y (USY particles 63-125 μm) was incubated with 5 mg / ml of benzalkonium chloride, after which the zeolite was allowed to drain and then dried overnight in a warm room. The zeolite with adsorbed benzalkonium chloride was repeatedly suspended at 20 mg / ml, first in PBS (■) and then in 95% ethanol (●). The concentration of benzalkonium chloride in the solution was measured by absorbance at 263 nm.
Example 7
This example shows the release of cephalothin from ultrastable zeolite (USY), which is described with reference to the chart in FIG.
[0039]
Ultrastable zeolite (USY) was incubated with the preservative cephalotin 5 mg / ml, after which the zeolite was allowed to drain before drying and further drying in a warm room. The zeolite with adsorbed cephalotin was repeatedly suspended at 20 mg / ml in 10 mM glycine pH 2.5 solution (■), and the concentration of cephalotin in the solution was measured by absorbance measurement at 260 nm. The zeolite was then suspended in a 10 mM phosphate buffer pH 8.0 (•). The change in pH caused cephalothin to deprotonate, and the charge introduced resulted in increased release of cephalothin from the zeolite.
Example 8
This example demonstrates the adsorption and release of various antibiotics. Ultrastable zeolite Y (USY) was incubated with various antibiotics and the amount absorbed by the zeolite was calculated. The zeolite was then dried, suspended at 20 mg / ml in buffer, and the concentration of released antibiotic in the solution was determined.
[0040]
[Table 3]

[0041]
Example 9
Dealuminated zeolite (DAY; 63-125 microns) was loaded with m-crelesol solution (10 mg / ml) with a zeolite content of 40 mg DAY / ml of m-cresol solution. E. Coli (CU1867, ATCC # 47092) suspension was mixed with unloaded zeolite and zeolite loaded with m-cresol to allow sedimentation for 5 minutes. The supernatant is removed from the zeolite precipitate and the remaining enclosed bacterial suspension (8 × 10⁶A zeolite with a colony forming unit (CFU) was incubated at room temperature. After 18 hours, the precipitate was resuspended and the number of CFU was determined after coating on LB-agar. The test was performed in buffer (0.3336 mg / ml monosodium phosphate, 0.7064 mg / ml disodium phosphate, 2 mg / ml glycine and 41 mg / ml mannitol) in growth hormone (GH, 5.5 mg / ml mannitol). ml) in the buffer. The distributions from the two tests are given below in CFU.
[0042]
[Table 4]

[0043]
Example 10
Dealuminated zeolite (DAY; 63-125 microns) was loaded with m-cresol (10 mg / ml) and the zeolite content of a 40 mg DAY / m-cresol solution. The E. Coli (CU1867, ATCC # 47092) suspension was mixed with unloaded zeolite and zeolite loaded with m-cresol, allowing the mixture to settle for 5 minutes. The supernatant is removed from the zeolite precipitate and the remaining enclosed bacterial suspension (8 × 10⁶Zeolites with colony forming units (CFU) were incubated at room temperature and at + 8 ° C., respectively. The test was performed in LB-medium, the precipitate was resuspended after 18 hours at room temperature and after 2.5 days at + 8 ° C., respectively, and after coating on LB-agar, The number was measured. The result is given below in CFU.
[0044]
[Table 5]

[Brief description of the drawings]
[0045]
FIG.
FIG. 1 schematically shows the intended equilibrium system of the invention under ideal conditions in the form of a box.
FIG. 2
FIG. 2 schematically shows the elution curve for zeolites initially loaded to high saturation.
FIG. 3
FIG. 3 schematically shows a zeolite bed in the form of a column that is long enough to allow a dosage gradient to be formed in the zeolite.
FIG. 4
FIG. 4 schematically illustrates various zeolite configurations associated with a single chamber.
FIG. 5
FIG. 5 schematically illustrates various zeolite configurations associated with the upstream and downstream chambers.
FIG. 6
FIG. 6 shows a chart related to the first embodiment.
FIG. 7
FIG. 7 shows a chart related to the second embodiment.
FIG. 8
FIG. 8 shows a chart related to the fifth embodiment.
FIG. 9
FIG. 9 shows a chart related to the sixth embodiment.
FIG. 10
FIG. 10 shows a chart related to the seventh embodiment.

Claims (72)

An arrangement for controlling the content or growth of microorganisms in a fluid, comprising: a) a zeolite having micropores loaded with an agent, wherein the agent has an affinity for the zeolite micropores; Comprising a zeolite having preservative properties and b) the fluid, wherein the zeolite and the surrounding fluid are at least partially in contact or arranged to be at least partially contacted. The fluid contains a therapeutically active compound or composition in a therapeutically effective amount and concentration, and the total amount of agent in the zeolite corresponds to the preservative level for the fluid An arrangement characterized by being greater than the amount to be performed. 2. The arrangement according to claim 1, wherein the micropores of the zeolite have a major cavity of uniform size of approximately 3-11 °. 2. The arrangement according to claim 1, wherein the zeolite comprises a powder or a particle mass. 2. The arrangement according to claim 1, wherein the zeolite comprises a compacted powder or particle compact structure. 5. The arrangement according to claim 4, wherein the bulk porosity of the structure is at least 20%, preferably at least 30%, and most preferably at least 35%, and preferably at most 80%, preferably at most. An arrangement characterized by 70%, and most preferably at most 65%. The arrangement of claim 1, wherein the zeolite comprises a hydrophobic zeolite, wherein the zeolite has the general structural formula (AlO ₂ ) _x (SiO ₂ ) _y and the y / x ratio is at least 15. An arrangement characterized by having a general structural formula, preferably greater than 100, preferably greater than 200, and most preferably greater than 1000. 2. The arrangement of claim 1, wherein the agent comprises or consists essentially of a molecule having a molar weight of less than 3000, preferably less than 2000, and most preferably less than 1500, Or an arrangement characterized by consisting of them. The arrangement of claim 1, wherein the agent comprises at least one preservative. 2. The arrangement of claim 1, wherein the agent comprises at least one preservative. 10. The arrangement according to claim 9, wherein the preservative is benzyl alcohol, benzalkonium chloride, cetrimide, cholbutol, chlorohexidine, chlorocresol, hydroxybenzoates, phenyl alcohol, phenoxy alcohol, phenylmercuric nitrite, chloramphenicol. , Phenol, cresol, especially m-cresol, and combinations thereof. 2. The arrangement according to claim 1, wherein the affinity of the agent for the zeolite is expressed as w / w amount of the agent in the zeolite at its maximum saturation, preferably at least 25%, and most preferably. Is an arrangement characterized by an agent affinity for the zeolite of at least 50%. 2. The arrangement according to claim 1, wherein the w / w amount of the agent in the zeolite is less than 100%, preferably less than 50%, more preferably less than 30%, and most preferably the amount corresponding to the maximum degree of saturation. Is less than 10%. 2. The arrangement according to claim 1, wherein the w / w amount of the agent in the zeolite is higher than 0.01%, preferably higher than 0.1%, and most preferably the amount corresponding to the maximum saturation. An arrangement characterized by being higher than 1%. The arrangement of claim 1, wherein the amount of agent loaded on the zeolite is adapted to provide release of the agent to the preservative concentration of the agent in the fluid when contacting the fluid. An arrangement characterized by that. 2. The arrangement of claim 1, wherein the zeolite forms a column that is long enough to provide substantially equilibrium concentration in the fluid after partially passing the column through the column. A method comprising: The arrangement of claim 1, wherein the fluid comprises a gas. The arrangement of claim 1, wherein the fluid comprises a liquid. 2. The arrangement according to claim 1, wherein the fluid comprises defined low-weight molecules having a defined affinity for the zeolite lower than the agent. . 19. The arrangement according to claim 18, wherein the affinity of the low weight molecule is at most 0.5 times, preferably at most 0.1 times, and most preferably at most 0.05 times that of the agent. An arrangement characterized by the following. 2. The arrangement of claim 1, wherein the fluid comprises high weight molecules as defined. 21. Arrangement according to claim 20, characterized in that the amount of high-weight molecules is greater than 0.01 mg / ml, preferably greater than 0.1 and most preferably greater than 1 mg / ml. 21. The arrangement according to claim 20, wherein the high-weight molecule comprises one selected from the group consisting of proteins, polypeptides, carbohydrates, nucleic acid sequences, lipids or mixtures thereof. 21. The arrangement according to claim 20, wherein said high weight molecule comprises at least one therapeutically active compound or composition. 2. The arrangement according to claim 1, wherein the fluid contains nutrients for microorganisms. An arrangement for controlling the content or growth of microorganisms in a fluid, comprising: a) a zeolite having micropores loaded with an agent, wherein the agent has an affinity for the zeolite micropores; Comprising a zeolite having preservative properties and b) the fluid, wherein the zeolite and the surrounding fluid are at least partially in contact, or arranged to be at least partially brought into contact. An arrangement wherein the fluid contains molecules that are larger than the micropores of the zeolite and have a defined affinity for the zeolite lower than the agent. 26. The arrangement according to claim 25, wherein said molecule comprises or forms part of a therapeutically active compound or composition. 27. The arrangement according to claim 26, wherein said therapeutically active compound or composition is present in said fluid in a medically active concentration. 26. An arrangement according to claim 25, characterized by any of the features of claims 1 to 24. A method for controlling the content or growth of a microorganism in a fluid, wherein the fluid comprises a therapeutically active compound or composition in a therapeutically effective amount and / or concentration, wherein at least one of the fluids comprises Part is contacted with a zeolite having micropores, the agent having affinity for the zeolite micropores and having the micropores loaded with an agent having preservative properties, and the agent is converted from the zeolite. Releasing the fluid to increase the content of the agent in at least a portion of the fluid. 30. The method of claim 29, wherein said contacting comprises maintaining said fluid in a substantially static relationship with said zeolite. 30. The method of claim 29, wherein said contacting comprises moving said fluid with respect to said zeolite. 32. The method according to claim 31, wherein the moving step comprises agitating the fluid and / or the zeolite. 32. The method of claim 31, wherein said transferring comprises passing the fluid over the zeolite. 32. The method of claim 31, wherein said transferring comprises passing said fluid through macropores in said zeolite. 35. The method of claim 34, comprising passing the fluid through a column of zeolite that is long enough to substantially provide an equilibrium concentration of agent in the fluid before or at the column outlet. Features method. 30. The method of claim 29, wherein said contacting comprises maintaining said fluid in a static relationship within macropores in said zeolite. 37. The method of claim 36, wherein a substantial preservative level of the agent in the fluid is maintained during the static relationship. 30. The method of claim 29, comprising increasing the content of the agent in the fluid to a preservative level in the fluid. 30. The method of claim 29, wherein the step of substantially increasing the content of agent in the fluid to an equilibrium level for the loaded zeolite. 30. The method of claim 29, wherein said contacting step comprises batch contacting between said fluid and said zeolite. 30. The method of claim 29, wherein the contacting step comprises intermittent contact between the fluid and the zeolite that are administered more than one time. 42. The method of claim 41, wherein the intermittent contact comprises maintaining at least a portion, preferably all, of the fluid dose in a static relationship within the macropores in the zeolite. And how. 30. The method of claim 29, wherein said contacting step comprises a continuous contact between said fluid and said zeolite. 44. The method of claim 43, comprising continuously or batch refilling the agent in the zeolite. 46. The method of claim 44, wherein said replenishing comprises contacting the zeolite after delivering an agent to the fluid. 46. The method of claim 44, wherein said replenishing step comprises the step of separating an agent from the fluid after contacting said zeolite and delivering it to said zeolite. 47. The method of claim 46, wherein said separating comprises extracting the agent by contacting the fluid with a second zeolite that is not loaded with or loaded with the agent. A method comprising: 30. The method of claim 29, wherein the method may feature any of the features of the preceding claims. A method for controlling the content or growth of microorganisms in a fluid, comprising a zeolite having micropores having an affinity for the zeolite micropores and an agent having preservative properties, wherein the micropores are used. Contacting at least a portion of the fluid with a loaded zeolite to release an agent from the zeolite to increase the content of the agent in at least a portion of the fluid, wherein the fluid comprises A process comprising molecules that are larger than the micropores of the zeolite and have a lower defined affinity for the zeolite than the agent. 50. The method of claim 49, wherein the fluid is a therapeutically effective amount and / or concentration of a therapeutically active compound or composition. 50. The method of claim 49, wherein any of the features of the preceding claims can be characterized. An arrangement for controlling the content or growth of microorganisms in a fluid, comprising: a) an upstream chamber or conduit, b) a downstream chamber or conduit, c) a bed, wherein the fluid is at least from the upstream chamber through the bed. A zeolite having micropores loaded with an agent, disposed between the upstream chamber and the downstream chamber in a manner that allows passage to a downstream chamber, wherein the agent has an affinity for the zeolite micropores. And a bed comprising a zeolite having preservative properties and d) an arrangement comprising a fluid present in at least one of the upstream chamber, the bed and the downstream chamber, wherein the bed is loaded with the zeolite. The amount of agent that has been adapted to release the agent to the preservative concentration of the agent in the fluid when contacting the bed. Arrangement, characterized in comprising at improved. 53. An arrangement according to claim 52, characterized by any of the features of the preceding claims. An arrangement for controlling the content or growth of microorganisms in a fluid, comprising: a) an upstream chamber or conduit, b) a downstream chamber or conduit, c) a bed, wherein the fluid is at least from the upstream chamber through the bed. A zeolite having micropores loaded with an agent, disposed between the upstream chamber and the downstream chamber in a manner that allows passage to a downstream chamber, wherein the agent has an affinity for the zeolite micropores. A bed comprising a zeolite having preservative properties, and d) comprising a fluid present in at least one of the upstream chamber, the bed and the downstream chamber, at least downstream of the bed. An arrangement characterized in that said fluid volume comprises a preservative concentration of said agent. 55. The arrangement of claim 54, wherein the arrangement may feature any of the features of the preceding claims. A method for controlling the content or growth of microorganisms in a fluid, comprising: a) providing a bed of zeolite, wherein the zeolite is a micropore and has an affinity for the zeolite micropore. Having micropores loaded with an agent having preservative properties and having macropores that allow the fluid to pass through the bed; and b) Passing at least a portion of the fluid through the bed in a manner that allows release of the agent from the zeolite to increase the content of the agent in at least a portion of the fluid to the preservative concentration of the agent. The method characterized by the above. 57. The method of claim 56, wherein the method may feature any of the features of the preceding claims. A hydrophobic zeolite having micropores, wherein the zeolite has the general structural formula (AlO ₂ ) _x (SiO ₂ ) _y and the y / x ratio is at least 15. The zeolite micropores are loaded with an amount of preservative corresponding to more than 10% of the maximum zeolite saturation for the agent and corresponding to an absolute amount of more than 1% w / w for the zeolite. Zeolite characterized by the improvement that it is. 59. The zeolite according to claim 58, characterized in that the zeolite is loaded with an amount of preservative which corresponds to more than 25%, preferably more than 35%, of the maximum zeolite saturation for the agent. Zeolite. 59. The zeolite of claim 58, wherein the zeolite is loaded on the zeolite in an absolute amount greater than 5% w / w. 59. The zeolite of claim 58, wherein the loaded zeolite is improved. 59. A zeolite according to claim 58, characterized by any of the features of the preceding claims. Use of a hydrophobic zeolite having micropores, wherein the zeolite has the general structural formula (AlO ₂ ) _x (SiO ₂ ) _y and the y / x ratio is at least 15; The use wherein the zeolite micropores are loaded with a preservative, wherein the fluid and the loaded zeolite are contacted to control the microbial content or growth of the fluid to control the preservative in the fluid. Use wherein the concentration is raised to a preservatively effective concentration. 64. Use according to claim 63, which may feature any of the features of the preceding claims. A method for controlling the content or growth of microorganisms in a fluid, comprising: a) providing a hydrophobic zeolite having micropores, wherein the zeolite has a y / x ratio of at least 15. Having the structural formula (AlO ₂ ) _x (SiO ₂ ) _y , wherein the zeolite micropores are loaded with a preservative; and b) contacting the fluid with the zeolite to form at least a portion of the fluid. Increasing the concentration of the preservative to a preservatively effective concentration. 66. The method of claim 65, wherein the fluid is kept in contact with the zeolite for a time sufficient for preservative action on microorganisms. 66. The method of claim 65, wherein the fluid is removed and a new portion of the fluid contacts the zeolite. 66. The method of claim 65, wherein any of the features of the preceding claims can be characterized. A method for controlling the content or growth of microorganisms in a fluid comprising the steps of: a) converting a first portion of the fluid to a zeolite having micropores loaded with an agent, wherein the agent has an affinity for the zeolite micropores. Contacting with a zeolite having a protective and preservative property to release the agent from the zeolite and increase the content of the agent in at least a portion of the first portion of the fluid; b) Maintaining a first portion of the fluid in static contact with the zeolite under preservative conditions; c) removing the first portion of the fluid from contact with the zeolite having a reduced agent content. And d) contacting a second portion of the fluid with a zeolite having the reduced agent content. 70. The method of claim 69, wherein any of the features of the preceding claims can be characterized. A method for controlling the content or growth of microorganisms in a fluid comprising the steps of: a) converting a first portion of the fluid to a zeolite having micropores loaded with an agent, wherein the agent has an affinity for the zeolite micropores. Contacting with a zeolite having a hydrophobic and preservative property to release the agent from the zeolite and increase the content of the agent in at least a portion of the fluid; b) removing the first portion of the fluid Removing from contact with the zeolite; c) separating at least a portion of the agent from a first portion of the fluid; e) adding at least a portion of the agent to a second portion of the fluid; d) contacting the second portion of the fluid with the zeolite. 73. The method of claim 71, wherein the method may feature any of the features of the preceding claims.

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