Lipid-based formulation for topical ophthalmic use to increase the bioavailability of non-steroidal Anti-inflammatory drugs in intraocular tissues

Abstract

The present invention relates to a lipid-based formulation for topical ophthalmic use to increase the bioavailability of non-steroidal anti-inflammatory drugs in intraocular tissues that comprises an effective amount of a non-steroidal anti-inflammatory drug (NSAID), polyethylene glycol (PEG)-12 glyceryl dimyristate, polyethylene glycol (15)-hydroxystearate (Kolliphor® HS15), an organic solvent, and excipients selected from the group consisting of buffering agents.

Description

[0001]LIPID-BASED FORMULATION FOR TOPICAL OPHTHALMIC USE TO INCREASE THE BIOAVAILABILITY OF NON-STEROIDAL ANTI-INFLAMMATORY DRUGS IN INTRAOCULAR TISSUES. FIELD OF THE INVENTION This invention relates to ophthalmic therapies, and more particularly to the use of a lipid base for the application of a fluid pharmaceutical formulation to ocular tissues for the local, targeted delivery of non-steroidal anti-inflammatory drugs. BACKGROUND OF THE INVENTION Ocular inflammation is considered to be associated with 10-15% of blindness cases, so its management is a priority for the prevention of visual loss.Topical corticosteroids have been used as drugs of choice in the management of ocular surface and anterior segment inflammation, while intravitreal corticosteroids are used in the management of intraocular inflammation; however, the association of topical corticosteroids and intravitreal corticosteroids with serious adverse effects has raised the possibility of increasing the use of other drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs), as well as developing safe platforms for delivering these active agents to the interior of the eyeball, with the purpose of increasing their bioavailability and therapeutic effect. Therefore, in order to influence the management of intraocular inflammation, it is necessary to develop safe pharmacological platforms that favor the penetration of NSAIDs into intraocular tissues, such as the presented invention.The eyeball can be divided into two parts: the anterior and posterior segments. The anterior segment is composed of the pupil, cornea, iris (part of the uvea), ciliary body, aqueous humor, and lens, while the posterior segment is composed of the vitreous humor, macula, retina, choroid (part of the uvea), and optic nerve. The retina is the layer of sensory tissue that creates nerve impulses that are transmitted through the optic nerve to the brain. The macula is a specialized area of ​​the retina responsible for central color and high-resolution vision. The different tissues of the anterior and posterior segments can suffer from inflammatory diseases of diverse etiologies, such as conjunctivitis, keratitis, uveitis, and macular edema, among others, which together are referred to as ocular inflammation.Ocular inflammation is the local response of ocular tissues and their adnexa to an attack of a wide variety of etiologies; it can cause irreversible damage to ocular function and its associated structures, potentially leading to blindness. Uveitis, along with other inflammatory ocular pathologies, has an incidence of 15 to 17 cases per 100,000 inhabitants per year in developed countries and accounts for 10% to 15% of new cases of blindness. Treatment of ocular inflammation can be targeted, targeting the etiology, or nontargeted, reducing inflammatory symptoms.Topical corticosteroids have been used as drugs of choice in the nonspecific treatment of anterior segment ocular inflammation, but their adverse effects, such as uncontrollable ocular hypertension leading to glaucoma and the induction of the appearance and / or progression of cataracts, have raised the need to employ other drugs such as nonsteroidal anti-inflammatory drugs (NSAIDs). The anti-inflammatory activity of NSAIDs lies in the reversible inhibition of the activity of the cyclooxygenase (COX) enzyme in its COX-1 and COX-2 isoforms, preventing the formation of prostaglandins, substances that mediate inflammation. NSAIDs also have an antiangiogenic action that is very useful for treating long-term inflammatory processes that are usually associated with the formation of new vessels (angiogenesis).On the other hand, intravitreal corticosteroids are preferred for the management of posterior segment inflammation. However, this route of administration is also associated with serious complications such as endophthalmitis (intraocular infection) and retinal detachment. NSAIDs are currently used in cataract surgery, in the prevention and treatment of cystoid macular edema, and for the short- and long-term treatment of allergic conjunctivitis. Among the NSAIDs marketed for ophthalmic use are ketorolac, nepafenac, diclofenac, and bromfenac. However, topical administration of NSAIDs in ophthalmic pathologies, particularly in inflammatory diseases of the posterior segment, is limited due to the low water solubility of these compounds. This low solubility prevents effective NSAID penetration and reduces their intraocular bioavailability.In order to increase the bioavailability of NSAIDs in intraocular tissues for their effective use in inflammatory ocular pathology, it is necessary to develop pharmacological platforms for the delivery of these active ingredients to the tissues of the anterior and posterior segments of the eye. The invention presented consists of a lipid-based formulation for topical ophthalmic use that functions as a liposomal delivery system for NSAIDs, particularly diclofenac, to tissues of the anterior and posterior segments of the eye. It is known that diclofenac is generally taken orally in the form of regular tablets or tablets coated with gastric juice-resistant coatings, or rectally, by injection, or topically.Oral administration of diclofenac is known to cause serious adverse effects such as gastrointestinal bleeding and ulceration, liver and kidney damage, and central nervous system and skin disorders, particularly after prolonged use. Therefore, in an effort to minimize the adverse effects associated with oral administration, non-oral delivery of diclofenac has been extensively investigated in recent years. Topical formulations are attractive options because they avoid hepatic first-pass metabolism, reduce side effects associated with oral administration, are associated with increased patient compliance, and, in some cases, improve the therapeutic efficacy of the drug. However, the efficacy of topical administration of diclofenac is limited by the difficulty of diffusing the drug and by its low solubility in water.This low solubility prevents effective penetration of diclofenac and reduces its intraocular bioavailability. Several patents and patent applications have attempted to address the aforementioned problems by developing topical gel formulations containing various ingredients to improve diclofenac solubility. US 4,711,906 describes a liquid diclofenac preparation, particularly for parenteral use, consisting of a solution of diclofenac or one of its salts and, if desired, additional pharmaceutical active ingredients and auxiliary substances in a solvent, the solvent consisting of 10-70% by weight, preferably 20-50% by weight, of a mixture of (a) propylene glycol and (b) polyethylene glycol and 90-30% by weight, preferably 80-50% by weight of water, and in the solvent mixture the weight ratio of propylene glycol:polyethylene glycol is between 9.5:0.5 and 0.5:9.5, preferably between 3:1 and 1:3, especially preferably between 2:1 and 1:2.US 4,917,886 describes a topically administrable pharmaceutical composition containing, as an active ingredient, from about 0.1 to about 10% by weight of a non-steroidal anti-inflammatory active compound, having at least one acid group, from about 10 to about 50% by weight of a volatile, water-soluble lower alkanol having from 2 to 4 carbon atoms, from about 3 to about 15% by weight of an optionally self-emulsifying lipid or a mixture of lipids, from about 0.5 to about 2% by weight of a gel structure former, from about 1 to about 20% by weight of a co-solvent, from about 40 to about 80% by weight of water, optionally about 0.5 to about 5% by weight of an emulsifier if the lipid phase is not self-emulsifying and, if desired, non-essential constituents.EP 147,476 describes a gel preparation for external application characterized in that it is prepared with diclofenac sodium as the active ingredient, water, lower alkanols and glycols as a medium, a carboxyvinyl polymer as a gelling agent and a weak basic substance as a neutralizing agent. The gel preparations for external application of this invention have good stability and pleasant feeling in use and show excellent anti-inflammatory and analgesic effects by skin absorption. EP 488,089 describes a diclofenac preparation for topical application that is packaged together with a propellant gas in a compressed gas container and can be foamed therefrom through an atomizer.EP 600,395 describes an anti-inflammatory and analgesic gel preparation comprising diclofenac or its salts, a dibasic acid ester, a lower alcohol, and a non-ionic polymer or a mixture of non-ionic polymers selected from the group consisting of (a) 1.5-4% by weight of hydroxypropylcellulose having a molecular weight of 500,000 or greater, (b) 2-4% by weight of hydroxyethylcellulose having a molecular weight of 1,250,000 or greater, and (c) 1.5-4% by weight of a mixture of hydroxypropylcellulose having a molecular weight of 500,000 or greater and hydroxyethylcellulose having a molecular weight of 1,250,000 or greater, and having a viscosity of 5,000-35,000 cps and a yield value of 5 dynes / cm2 or greater.EP 788,794 describes an externally applied composition, such as a liquid preparation, cream, ointment, or poultice plaster, characterized by a composition containing a water-soluble salt such as diclofenac sodium, water, and a fatty acid dialkylamide and / or its polyoxyethylene adduct. US application No. 2010 / 0016395 A1 reports being able to increase the dose of pilocarpine to 1% to 2% by adding the nonsteroidal anti-inflammatory agent diclofenac, but at a concentration that was five times more concentrated than that approved by the FDA. Congdon et al. reported that diclofenac is associated with serious side effects such as persistent epithelial defects, corneal melting, and corneal perforation. (Congdon et al., 2001, Corneal complications associated with topical ophthalmic use of nonsteroidal anti-inflammatory drugs, Ophthalmology, 27:622-631).WO 2009 / 077736 discloses a combination of pilocarpine and dapiprazole (or thymoxamine) and pilocarpine and brimonidine (or Iopidine) for treating visual acuity defects, presbyopia, myopia, hyperopia, reduced night vision, and astigmatism. The combinations indicated are pilocarpine and dapiprazole (or thymoxamine), and pilocarpine and brimonidine (or Iopidine). The combination of pilocarpine and dapiprazole produced ocular redness and irritation (WO 2009 / 077736), and it is known to those skilled in the art that topical administration of brimonidine produces lightheadedness, dizziness, dry mouth, rapid heart rate, and stomach upset, among other side effects, which limits its use even in patients using it for a serious ocular condition such as glaucoma.Novartis PCT patent publication WO2012 / 093101, entitled “Indole compounds or analogues thereof useful for the treatment of age-related macular degeneration,” describes certain Factor D inhibitors. Additional Factor D inhibitors are described in Novartis PCT patent publications WO2014 / 002051, WO2014 / 002052, WO2014 / 002053, WO2014 / 002054, WO2014 / 002057, WO2014 / 002058, WO2014 / 002059, WO2014 / 005150, and WO2014 / 009833. Bristol-Myers Squibb PCT patent publication WO2004 / 045518, entitled “Open-chain prolyl urea-related modulators of androgen receptor function,” describes open-chain prolyl urea- and thiourea-related compounds for the treatment of androgen receptor-associated conditions, such as age-related diseases, e.g., sarcopenia. Japan Tobacco Inc. PCT patent publicationWO1999 / 048492, entitled “Amid derivatives and antagonists of nociceptin,” describes compounds with a proline-like nucleus and aromatic substituents connected to the proline nucleus via amide bonds, useful for the treatment of pain. Alexion Pharmaceuticals PCT patent publication WO1995 / 029697, entitled “Methods and compositions for the treatment of glomerulonephritis and other inflammatory diseases,” describes antibodies directed against C5 of the complement pathway for the treatment of glomerulonephritis and inflammatory conditions involving pathological activation of the complement system. Alexion Pharmaceutical’s anti-C5 antibody eculizumab (Soliris®) is currently the only complement-specific antibody on the market, and is the first and only approved treatment for paroxysmal nocturnal hemoglobinuria (PNH).The prior art describes nonsteroidal anti-inflammatory agents (NSAIDs) as inhibitors of the enzyme cyclooxygenase, which produces prostaglandins. This enzyme has two forms: cyclooxygenase-1 (COX-1), which is thought to have both “resident” and “housekeeping” functions, and cyclooxygenase-2 (COX-2), which is upregulated in inflammation and cancer. Agents that selectively inhibit COX-2, as opposed to both COX-1 and COX-2, are thought to block inflammation without affecting the body's normal homeostatic mechanisms. (Fitzgerald GA and Patrono C. The coxibs, selective inhibitors of cyclooxygenase-2. NEJM 2001;345:433-442). Although molecules for the medical treatment of inflammatory diseases have been disclosed, no clinically effective preparation suitable for use by the general public without unreasonable side effects has been found.The invention is useful for treating multiple intraocular inflammatory diseases by increasing the bioavailability of non-steroidal anti-inflammatory drugs. Unlike previous compositions disclosed in the prior art, the compositions of the present invention can be safely used by patients for the treatment of various ocular conditions. The present invention aims to generate an innovative lipid-based pharmacological platform for topical ophthalmic use that increases the bioavailability of NSAIDs, particularly diclofenac, in intraocular tissues, thereby impacting the pharmaceutical industry in ophthalmology, an active and highly productive sector locally, nationally, and internationally.OBJECT OF THE INVENTION An object of the present invention is to propose a lipid-based formulation for topical ophthalmic use to increase the bioavailability of nonsteroidal anti-inflammatory drugs in intraocular tissues. Another object is to propose a pharmaceutical formulation comprising an aqueous solution of diclofenac for topical application that has improved diffusion and bioavailability properties. DESCRIPTION OF THE INVENTION Additional objects and advantages of the invention will be set forth in part in the description that follows, and in part will be obvious from the description, or may be learned by practice of the invention. It should be understood that the following detailed description is only exemplary and explanatory and does not limit the invention as claimed. In order that the present invention may be more readily understood, certain terms are defined. Other definitions are set forth throughout the description of the embodiments.The present invention relates to a lipid-based formulation for topical ophthalmic use to increase the bioavailability of nonsteroidal anti-inflammatory drugs in intraocular tissues. The formulation comprises an effective amount of a nonsteroidal anti-inflammatory agent (NSAID), a medium-chain fatty acid, an organic solvent, and excipients selected from the group consisting of buffering agents. Wherein said effective amount of a nonsteroidal anti-inflammatory agent (NSAID) refers to an aqueous solution of a pharmaceutically acceptable salt of diclofenac. The compositions of the present invention contain an effective amount of diclofenac (D), 0.001 to 10 mg, i.e. 0.0001 to 1% (w / v), polyethylene glycol (PEG)-12 glyceryl dimyristate which is used as a structural component of the liposomes in a concentration of 5-15% (w / v) and ethyl alcohol which is used as an organic solvent in a concentration of 0.7 to 2.1% (v / v).In addition, the formulation contains 2.5 to 7.5% (w / v) of polyethylene glycol (15) hydroxystearate (Kolliphor® HS15) as a non-ionic solubilizer and emulsifying agent. This compound is characterized by its low toxicity and also acts as a permeability enhancer, because it promotes the transport of molecules across cell membranes; increasing the rate of endocytosis and stimulating the translocation of drugs through the paracellular route. Likewise, the aqueous compositions of the present invention optionally comprise further excipients selected from the group consisting of buffering agents, pH adjusting agents and preservatives. Anhydrous citric acid (0.04 to 0.16%) and sodium citrate dihydrate (0.23 to 0.69%) are used as buffers. The lipid-based compositions of the present invention can be prepared by conventional methods for preparing suspension pharmaceutical compositions.It was surprisingly found that the compositions of the present invention enhance the action and decrease the side effects. Method of preparation. According to the preferred method, diclofenac powder or solution (concentration 0.001 to 10 mg) is first added to a lipid mixture containing polyethylene glycol (PEG)-12 glyceryl dimyristate (5-15%), polyethylene glycol (15)-hydroxystearate (Kolliphor® HS15) 2.5 to 7.5% and ethyl alcohol (0.7 to 2.1%), the components being combined by stirring for 10 minutes ^1 minute at 25 ° C^ 1 ° C. On the other hand, an aqueous base mixture composed of grade 2 purified water (QS), anhydrous citric acid (0.04 to 0.16%) and sodium citrate dihydrate (0.23 to 0.69%) is prepared by combining the components by stirring for 10 minutes ^1 minute at 25 ºC^1 ºC. The aqueous mixture is added to the lipid mixture at room temperature and stirred for 5 min ^1 minute, thus obtaining the final formulation. Resulting formulation.The resulting lipid-based formulation object of this invention is formed as follows: Reagent Volume / Amount Diclofenac 0.001 to 10 mg (w / v) Polyethylene glycol (15) -hydroxystearate 2.5 to 7.5% (w / v) Polyethylene glycol (PEG)-12 glyceryl dimyristate 5 to 15% (w / v) Ethyl alcohol 0.7 to 2.1% (v / v) Anhydrous citric acid 0.04 to 0.16% Sodium citrate dihydrate 0.23 to 0.69% Benzalkonium chloride 0.1 mg Purified water grade 2 QS1.0 ml These and other embodiments of the invention can be further illustrated in the following non-limiting examples. EXAMPLE. The following example is intended to illustrate, but not to limit, the present invention [ingredient concentrations are presented in units of % weight / volume (% w / v) or % volume / volume (% v / v)]. The final concentration of diclofenac in the resulting nano suspension in the example is 1 mg / ml (0.1%).The composition of the lipid emulsion (L) with diclofenac (D), hereinafter referred to as LD, is described below. REAGENT VOLUME / QUANTITY Diclofenac 1 mg Polyethylene glycol (15) hydroxystearate 75 mg Polyethylene glycol (PEG)-12 glyceryl dimyristate 100 mg Ethyl alcohol 14 µl Anhydrous citric acid 0.8 mg Sodium citrate dihydrate 4.675 mg Benzalkonium chloride 0.1 mg Purified water grade 2 QS1.0 ml Analysis of the resulting formulation. The pH of the LD was analyzed by a potentiometer in triplicate at room temperature. The osmolarity was measured by a vapor pressure osmometer in triplicate at 33°C (the ocular surface temperature). The viscosity was also measured in triplicate at 33°C. Viscosity was measured using a thermostatically controlled rheometer when steady state was reached with shear rates increasing from 0 to 1000 s-1.The particle size of the diclofenac-containing lipid nanoparticles in the DL was analyzed by dynamic light scattering, and the zeta potential (ζ) was calculated by measuring the particle velocity using a laser Doppler velocimeter at 25°C (Zetasizer Nano ZS, Malvern Instruments, Malvern, UK). The particle size distribution (Z) and the polydispersity index (PDI) were calculated from the particle size distribution. The pH of the DL was 7.4. The physiological pH range of tears is between 6.5 and 7.6. The viscosity of the DL was 36.5 cP (high), which could favor its permanence on the ocular surface. The osmolarity was 367 mOsmol / L, within the isotonic range. The average particle diameter (z-average) was 152.45 nm, while its PDI was 8.78 and its zeta potential was 14.25. The physicochemical characteristics of the LD are summarized below.PHYSICOCHEMICAL CHARACTERISTICS OF LD. Formulation Physicochemical parameters pH Viscosity Osmolarity Z PDI (cP) (mOsm / l) (nm) (nm) LD 7.4 36.5 367 152.45 8.78 pH= Hydrogen ion potential, cP= centipoise, PDI= Polydispersity index Additionally, diffusion chambers and New Zealand white rabbit corneas were used to perform in vitro diffusion experiments (BW200S Chemotaxis Chambers, NeuroProbe, Gaithersburg, MD, USA) in order to test the formulation’s ability to increase the bioavailability of diclofenac. Briefly, the central section of the cornea (6 mm in diameter) was placed between the upper and lower compartments of the diffusion chambers to act as a diffusion barrier for either the LD or a control solution of diclofenac in aqueous solution (AD), composed of the active ingredient (1 mg / mL) in water for injection.The upper compartment (acceptor chamber) was filled with 180 μL of balanced salt solution (BSS), while the lower compartment (donor chamber) was filled with 200 μL of either LD or AD. To prevent evaporation, the diffusion chambers were placed inside a humidified chamber at 37 °C. To analyze LD diffusion, the concentration of diclofenac (D) was determined by high-performance liquid chromatography (HPLC) in the upper compartment at 2, 4, 6, and 8 hours after the start of the diffusion test. The determination of D concentration by HPLC was carried out using the Varian 920 LC equipment (Aligent Technologies, Santa Clara, CA, USA) with a Zorbax Eclipse Plus C18 column, 4.6 x 100 mm and 3.5 μm column (Agilent, Santa Clara, CA, USA) at a temperature of 30 °C. The samples (20 μl) were eluted from the column in a mobile phase composed of water:methanol (30:70) at a flow rate of 1 ml / min.Detection was performed at 225 nm. The retention time and detection limit were 5 min and 75 - 4000 ng / mLml, respectively. Regarding the in vitro diffusion analysis, it was observed that the LD reached higher concentrations of D compared to AD. The results of the in vitro diffusion study are presented below. DICLOFENAC (D) CONCENTRATION IN THE UPPER COMPARTMENT OF THE DIFFUSION CHAMBER (NG / ML). Time (hours) LD AD 2 834.92 562.23 4 948.57 745.83 6 1450.31 890.78 8 1470.87 910.61 The values ​​represent the average of three measurements. The D concentrations in the acceptor chamber were higher with the LD. which reflects the invention's ability to significantly increase the deposition of the NSAID (D) in intraocular compartments and tissues. It has been surprisingly found that the pharmaceutical formulation of the present invention has improved diffusion and bioavailability properties.It has also been found that the pharmaceutical formulation of the present invention is stable, and can be stored throughout the shelf life of the product without any separation or precipitation of free diclofenac from solution. In order to further define the invention, the following terms and definitions are provided herein: DEFINITIONS As used herein, a “composition” refers to a material suitable for administration to an eye of a subject. Compositions may include a drug delivery system if desired. Compositions may comprise a liquid carrier. The term may also be used to refer to materials such as solutions, suspensions, emulsions, and the like.The term “therapeutically effective amount” as used herein refers to the level or amount of agent necessary to treat an ocular condition without causing significant adverse or negative side effects to the eye or a region of the eye. The term “pharmaceutically acceptable” as used herein means suitable for use in contact with tissues without undue toxicity, irritation, incompatibility, instability, allergic response, and the like. The term “% (w / v)” used herein means parts by weight (expressed in grams) per 100 parts by volume (expressed in milliliters). Thus, an aqueous solution containing, for example, 1% w / v diclofenac means that 100 ml of aqueous solution contains 1 gram of diclofenac.DICLOFENAC Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID) chemically known as 2-[(2,6-dichlorophenyl)amino]phenylacetic acid. Diclofenac belongs to the acetic acid class of NSAIDs. The drug was developed in the 1960s by scientists at Ciba-Geigy and is sold worldwide by Novartis under several trade names, including Cataflam™ and Voltaren™ in Europe and the United States. Due to its excellent analgesic characteristics, diclofenac is widely used to treat various types of pain, including chronic and acute painful episodes.The drug is used to treat musculoskeletal and joint disorders such as rheumatoid arthritis, osteoarthritis, and ankylosing spondylitis; periarticular disorders such as bursitis and tendonitis; soft tissue disorders such as sprains and strains; and other painful conditions such as after certain surgical procedures. Diclofenac is usually taken orally in the form of plain tablets or tablets coated with gastric juice-resistant coatings, or rectally, by injection, or topically. Oral administration of diclofenac can cause serious adverse effects such as gastrointestinal bleeding and ulceration, liver and kidney damage, and central nervous system and skin disorders, particularly after prolonged use.Advantageously, the pharmaceutical formulation of the present invention comprises a diclofenac salt selected from the sodium, potassium, pyrrolidine, piperidine, N-hydroxyethylpyrrolidine, N-hydroxyethylpiperidine, triethanolamine, diethanolamine, ethylenediamine, and diethylamine salts of diclofenac. While only certain features of the invention have been illustrated and described herein, many modifications and changes will occur to those skilled in the art. It is, therefore, to be understood that the appended claims are intended to cover all modifications and changes which fall within the true spirit of the invention.

Claims

CLAIMS Having sufficiently described my invention, I consider as a novelty and therefore claim as my exclusive property, the contents of the following clauses: 1.- A lipid-based formulation for topical ophthalmic use to increase the bioavailability of non-steroidal anti-inflammatory drugs in intraocular tissues characterized in that it comprises a combination of diclofenac, polyethylene glycol (PEG)-12 glyceryl dimyristate which is used as a structural component of the liposomes and ethyl alcohol which is used as an organic solvent, polyethylene glycol (15)-hydroxystearate (Kolliphor® HS15) as a non-ionic solubilizer and emulsifying agent, medium chain fatty acids. 2.- The lipid-based formulation for topical ophthalmic use according to claim 1, characterized in that it comprises the following percentages: Diclofenac ------------------------------------------------- 0.001 to 10 mg (w / v); Polyethylene glycol (15) hydroxystearate ----------------- 2.5 to 7.5% (w / v); Polyethylene glycol (PEG)-12 glyceryl dimyristate -- 5 to 15% (w / v); Ethyl alcohol ---------------------------------------------- 0.7 to 2.1% (v / v); Anhydrous citric acid ------------------------------------- 0.04 to 0.16%; Sodium citrate dihydrate ------------------------------- 0.23 to 0.69%; Benzalkonium chloride --------------------------------- 0.1 mg; Purified water grade 2 -------------------------------- QS1.0 ml.