Ethanolic extract to counter borrelia spp.-induced diseases such as lyme borreliosis and relapsing fever

Abstract

The invention relates to an ethanolic extract comprising sage extracts and to a method for the production of said extract.

Description

[0001] Ethanolic extract against diseases caused by Borrelia spp., Lyme borreliosis and

[0002] Relapsing fever

[0003] According to the invention, an ethanolic extract comprising sage extracts is proposed, as well as a method for its production.

[0004] Tick-borne encephalitis (TBE) and tick-borne encephalitis (TME) were first described in the 1980s, bringing tick-borne diseases into the public eye for the first time. Tick-borne encephalitis is a disease caused by the TBE virus that presents with symptoms similar to Lyme disease, namely fever and, in some cases, meningoencephalitis, an inflammation of the brain and meninges [1].

[0005] State of the art:

[0006] [1]

[0007] Sebastian Wendt et al (2019): Tick-borne diseases: From Lyme borreliosis to Q fever and tick-borne encephalitis. CME Volume 16 (5): 53-71

[0008] [2]

[0009] Hartmut Krauss et al. (2004): Lyme disease. In: Hartmut Krauss, Albert Weber, Burkhard Enders (eds.): Zoonoses. Infectious diseases transmissible from animals to humans. 3rd edition. Deutscher Ärzteverlag. Cologne.

[0010] [3]

[0011] Fukai Bao et al (2021 / 2022): Global seroprevalence and sociodemographic characteristics of Borrelia burdorferi senso lato in human populations: a systematic review and meta-analysis. BMJ Global Health Volume 7 Issue 6.

[0012] [4]

[0013] Dirk Weickmann, Toximed GmbH (2006): Pharmaceutical active ingredient for the treatment of hepatitis, Lyme disease and multiple sclerosis. European Patent Office EP 1754486A3

[0014] [5]

[0015] Boss. & Heldr. by Benth. (1848): Salvia pisidica. APde Candolle, Prodr. 12:269.

[0016] [6] Gülcan Özkan, Sevil Albayrak (2010): Study on chemical composition and biological activities of essential oil and extract from Salvia pisidica. LWT - Food Science and Technology Volume 43, Issue 1, Pages 186-190.

[0017] [7]

[0018] Renate Burda, Jenny Mörth-Kretschmer, Elisabeth Sand, Dirk Weickmann, Patentpool Target GmbH (2022): Kit of Parts zur Therapie der durch Borrelia spp. ausgelösten Borreliose. European Patent Office EP 4233 547 A1

[0019] [8]

[0020] Akgul H, Korkmaz N, Dayangac A, Sevindik M (2020): Antioxidant Potential of Endemie Salvia absconditiflora. Turkish Journal of Agriculture - Food Science and Technology, 8(10), 2222-2224

[0021] [9]

[0022] Janicsak G, Zupko I, Hohmann J (2011): Bioactivity-guided Study of Antiproliferative Activities of Salvia Extracts. Natural Product Communications, Vol. 6, No. 5, 575-579

[0023]

[0010]

[0024] Özcelik B, Erdogan Orhan I, Kan Y (2011): Determination of Antiviral Activity and Cytotoxicity of Selected Sage (Salvia L.) Species. FABAD J. Pharm. Sei., 36, 155-160, 2011

[0025]

[0011]

[0026] Su CY, Ming QL, Rahman K, Han T, Qin LP. (2015): Salvia miltiorrhiza: Traditional medicinal uses, chemistry, and pharmacology. Chin J Nat Med. 2015 Mar; 13(3): 163-82

[0027]

[0012]

[0028] Lin TH, Hsieh CL (2010): Pharmacological effects of Salvia miltiorrhiza (Danshen) on cerebral infarction. Chin Med 5, 22 (2010)

[0029] Lyme disease is caused by bacteria, unlike tick-borne encephalitis (TBE), which is caused by a virus. Back then, the risk of transmission was only one in every 10,000 or 100,000 tick bites. Nowadays, it can be assumed that Borrelia bacteria are transmitted with every tick bite. The danger posed by Borrelia and its connection to rheumatic diseases went unrecognized for a long time, partly because the incubation period between a tick bite and the onset of the disease can be up to 40 years. In most cases, months or even years pass after a tick bite before the disease manifests. Presumably due to the generally weakened immune systems of the modern population and the widespread environmental pollution, the disease now often develops much more quickly and is more severe.We are convinced that most rheumatic complaints that occurred in the past, and still occur today, especially among the rural population, are actually due to Borrelia infections [2], In addition to Lyme borreliosis, which is caused by the Borrelia species:.

[0030] - Borrelia burgdorferi

[0031] - Borrelia afzelii

[0032] - Borrelia garinii

[0033] - Borrelia bavariensis

[0034] - Borrelia mayonii

[0035] - Borrelia spielmanii

[0036] While Borrelia texanensis may be the cause, there is another disease that can be caused by Borrelia bacteria and transmitted by ticks: relapsing fever. The pathogens were first identified in 1868 by the German physician and bacteriologist Otto Obermeier (1843-1873) in the blood of patients. The species identified was Borrelia recurrentis. It is now known that Borrelia duttoni, B. crocidurae, B. hispanica, B. hermsii, B. venezuelensis, B. turicae, B. mazzottii, B. persica, and B. parkeri can also cause relapsing fever.

[0037] Lyme disease initially manifests with nonspecific general symptoms such as fever, headache, or gastrointestinal problems. A characteristic skin rash (erythema migrans or bull's-eye rash) is a clear symptom of a Lyme disease infection, but it does not occur in all cases. The absence of erythema migrans after a tick bite does not automatically rule out a Lyme disease infection.

[0038] Symptoms such as flu-like symptoms, fever, and headaches make the disease difficult to recognize. It can affect organs, joints, muscles, and the central and peripheral nervous systems. At this stage, the immune system is often no longer able to fight the infections. Borrelia bacteria remain in the bloodstream only briefly and attach themselves to connective tissue and cell walls, essentially living hidden. Here, they are difficult to detect, necessitating prolonged treatment. Relapsing fever is characterized by repeated episodes of high fever mediated by cytokines. Furthermore, the pathogens can damage the vascular endothelium, leading to an increased tendency to bleed and necrosis of the affected organs. If the pathogen is not completely eliminated, sepsis and multiple organ failure ultimately result, leading to the patient's death.

[0039] Other symptoms such as joint, head, chest and limb pain, as well as nausea and shortness of breath, can closely resemble those of Lyme disease.

[0040] State of the art in treatment:

[0041] If the infection is not yet advanced, it can be treated with antibiotics. Treatment with pure penicillin (C16H18N2O4S) has proven most effective. Whether other antibiotics have a lasting and sustainable effect is currently unconfirmed and remains questionable. However, penicillin treatment can be helpful if the right moment (before the bacteria "hide" in the tissue) is recognized; however, recognizing this moment is virtually impossible. From the time of transmission of the pathogen, a local skin infection can occur after a varying incubation period, manifesting as redness at the bite site. The incubation period can last several years in some cases, which is why the associated joint pain was long mistaken for rheumatic complaints and unsuccessfully treated with cortisone ointment.We are convinced that most rheumatic complaints, especially those that occurred in the past among the rural population and still occur today, are due to Borrelia.

[0042] Problems with diagnosis and therapy:

[0043] Contrary to popular belief, Borrelia bacteria can spread very quickly. Lyme disease does not have a completely predictable course, as the classic three-stage classification suggests; its progression is not linear. If a patient's symptoms do not fit the conventional medical classification of Lyme disease, it is often ruled out, and the symptoms are treated incorrectly. A Lyme disease infection does not follow any pattern! The development of Lyme disease cannot be predicted. Symptoms can develop rapidly and severely, or they can manifest gradually and slowly. If Lyme disease is not treated promptly, it can become a chronic infection. This means the disease recurs repeatedly or worsens progressively. Even years of symptom-free latency followed by reactivation are possible.The pathogens cause various illnesses. The joints are most commonly affected, but heart problems and vascular inflammation can also occur. Mixed forms of the disease are also possible. Many Lyme disease patients complain of exhaustion and chronic fatigue that cannot be alleviated by sufficient sleep. A Lyme disease vaccine suitable for humans is not expected in the near future. In the USA, a corresponding vaccine was withdrawn from the market in 2002 due to alleged vaccination complications. However, this vaccine was only effective against the American strain of the pathogen and therefore not applicable in Europe. The only truly effective antibiotic is purely biological penicillin. According to our experience, this must be administered no later than six weeks after infection. This only happens in very few cases.

[0044] Relapsing fever usually has an acute course. Penicillin has presented problems in this context, as its use can regularly lead to relapses.

[0045] A risk inherent in the treatment of Borrelia is the so-called Jarisch-Herxheimer reaction. This can occur if too many Borrelia bacteria are killed at once with antibiotics, and the bacterial endotoxins released in the process can cause circulatory shock in the treated patients, which can sometimes be fatal.

[0046] Lyme disease update:

[0047] Recent studies show that approximately one in seven people worldwide has been infected with Lyme disease at least once. According to a study published in BMJ Global Health, Central Europe has the highest infection rate at 20% [3]. For relapsing fever, there are no reliable data sources. However, it can be assumed that approximately one in 15 to 20 people worldwide has been infected at least once.

[0048] The objective of this invention is therefore to develop an effective therapy against Borrelia species that can cause Lyme disease and / or relapsing fever. With patent W02007060252A3, we have already had a therapeutic agent against Lyme disease since 2006, which has also been positively evaluated within the framework of permissible medical treatment trials. Here, we identified and developed active ingredients from the total toxin cocktails of various Myriapoda [4]. A disadvantage of this treatment option is the relatively poor availability of the active ingredients. With patent EP 4233 547 A1 [7], we presented three sage species that can kill Borrelia burgdorferi and / or Borrelia afzelli. This is effective, but does not fully suffice as a therapy against the diseases caused by Borrelia, namely Lyme disease and relapsing fever.It is known from the literature that various sage species may be used against tumor diseases [8, 9], and it is also assumed that viruses can be combated by means of the essential oils of various sage species

[0010] . An application against various human pathogenic bacteria is not known, especially not against Lyme disease. For this purpose, there is only the method from TCM that red root sage (Salvia miltiorrhiza) can be used against Lyme disease

[0011] .

[0049] In the present invention we describe an effective treatment option for Lyme borreliosis and relapsing fever with best availability of the active ingredients and optimal tolerability of the therapy for the patient.

[0050] Inventive composition and manufacture:

[0051] The following describes aspects of the present invention.

[0052] We have now surprisingly found substances in several sage species [5] that can kill Borrelia spp. and Yersinia spp. in vitro and in vivo. It is currently assumed that a methanol extract, or the oil of various Salvia spp., exhibits antimicrobial effects against Gram-positive bacteria. Hardly any effect has been demonstrated against Gram-negative bacteria. Our experiments to date confirm this current scientific understanding.

[0053] We have now found that an ethanol extract of the fresh leaves of the Salvia spp. listed in Table A, when incorporated into honey, can kill gram (-) bacteria, specifically Borrelia spp. and / or Yersinia spp. [6],

[0054] Therefore, an ethanolic extract of the Salvia spp. we examined, incorporated into honey, is suitable for the treatment of Lyme disease and relapsing fever.

[0055] Approximately 900 species of sage are known worldwide, and there are likely well over 1,000 species. Many contain substances of pharmaceutical and medical interest. Only a few species have been studied to date. We investigated extracts of the following sage species for their antiborreliosis properties:

[0056] - Salvia mellifera

[0057] - Salvia absconditiflora

[0058] - Salvia caespitosa - Salvia tomentosa

[0059] - Salvia taraxacifolia

[0060] - Salvia nevadensis

[0061] - Salvia fruticosa - Salvia jurisicii

[0062] - Salvia recognita

[0063] - Salvia cinerea

[0064] - Salvia pisidica

[0065] - Salvia miltiorrhiza - Salvia lavandulifolia

[0066] - Salvia officinalis Krk

[0067] - Salvia officinalis Dalmatia

[0068] - Salvia azurea

[0069] Table 1: Efficacy of sage extracts against different Borrelia spp.

[0070] Only extracts of Salvia absconditiflora, Salvia caespitosa, Salvia recognita, Salvia tomentosa, Salvia jurisicii, and Salvia pisidica showed a demonstrable effect against Borrelia according to the invention. Salvia cinerea, Salvia nevadensis, Salvia taraxacifolia, and Salvia miltirrhoriza were also active against individual Borrelia species.

[0071] According to the invention, a mixture of different extracts makes sense and should be combined according to the respective clinical picture.

[0072] The following Salvia spp. are preferred, or particularly preferred, as extracts for the application according to the invention, either alone or in combination:

[0073] Table A: The sage species that can be used according to the invention

[0074] The composition according to the invention is prepared in the following manner: a)

[0075] A specific quantity of leaves, preferably fresh leaves of Salvia spp., is introduced into a specific quantity of 96% ethanol, or preferably 100% ethanol. b)

[0076] The extract from a) is left to stand for at least 24 hours at temperatures preferably of 25 degrees Celsius. The leaves are then filtered out. c)

[0077] This active extract from b) is incorporated into 150 mL of liquefied organic blossom honey, preferably organic rosemary honey, while stirring constantly.

[0078] Cultivation of Salvia spp. for optimal active ingredient synthesis:

[0079] In order for plants, especially the species Salvia absconditiflora, Salvia tomentosa, Salvia recognita, Salvia pisidica, and Salvia caespitosa required for the use according to the invention, to produce relevant quantities of their active ingredients, it is important to have the right growing media / soils for the plants. We found that for the Salvia spp. according to the invention, not only is permeable soil necessary, but that the species require calcareous soil for active ingredient synthesis. We preferably use calcareous soil from flora montana with approximately 20%–35% weathered limestone.

[0080] More than 35 different components can be detected in the ethanol extract of the Salvia spp. we used. Bile acid equivalents, rutin equivalents, and catechin equivalents can be increased by using the correct substrate. We were also able to detect peptide toxin-like substances that have not yet been chemically described. Cultivation in plant containers is more efficient than cultivation directly in the soil.

[0081] For the application according to the invention, it is possible to use the fresh leaves of the plants without the stems. We observed effects according to one aspect of the present invention at the following concentrations:

[0082] Therefore, the higher concentrations and the extracts in 100% ethanol are the most effective.

[0083] Dosage forms:

[0084] It has been shown that the extracts can be taken orally. The active ingredients are absorbed via the digestive tract. The ethanolic extract dissolves particularly well in honey, especially rosemary honey; therefore, in addition to taking the ethanolic extract directly and undiluted, taking it dissolved in honey, preferably organic rosemary honey, is also advisable.

[0085] Within the scope of the present invention, young leaves are defined as leaves that are a maximum of 6 days old, preferably a maximum of 4 days old. The age is counted from the beginning of the opening of the leaf bud. This is advantageous because the highest concentrations of pharmaceutical active ingredients are found in young leaves. This characteristic protects the young leaves from herbivores and pathogens during their early development. Within the scope of the present invention, fresh leaves are defined as leaves that have been harvested, i.e., typically cut, for a maximum of 24 to 72 hours.

[0086] This is advantageous because after harvesting the leaf undergoes chemical and physical changes, and the preferred properties then decrease.

[0087] According to one aspect of the present invention, allowing the extract to stand for at least 24 hours means that it remains untreated for at least 24 hours. This is advantageous because the highest concentration of the extract with the desired ingredients occurs within 24 hours.

[0088] It is advantageous to stir the extract approximately every two hours. Stirring means gently agitating the surface, preferably just enough to alter it. This creates a homogeneous mixture that, in turn, facilitates further extraction of the active ingredients. This is preferably done counterclockwise, which has the advantage of improving the extraction of left-handed active ingredients.

[0089] Approximately 25 degrees Celsius, according to one aspect of the present invention, means a preferred temperature of 24 to 26 degrees Celsius. This is advantageous because the most effective active ingredient extraction occurs at this temperature. This has been empirically demonstrated.

[0090] For liquefied organic blossom honey, 150 ml can be used to achieve standardization. This is advantageous because it allows for a one-month supply of the extract to be prepared for therapeutic trials. This is beneficial because the extract remains stable for approximately one month without significant change.

[0091] According to one aspect of the present invention, but optionally, blossom honey and / or rosemary honey are provided in organic quality. Organic quality means that the honey is low in pollutants and toxins. This is advantageous because the extract is used in people with health problems, thus reducing or virtually eliminating further exposure.

[0092] At least 96% ethanol means that ethanol with a volume fraction of at least 96% is provided, i.e. approximately 96%, 97%, 98%, 99% or 100% volume fraction.

[0093] This is advantageous because the best solubility is achieved with ethanol of 96% or higher.

[0094] "Combating Lyme disease" means that the concentration of Borrelia bacteria in blood plasma is at most 1%. The symptoms therefore subside or preferably disappear completely. The Borrelia bacteria, the causative agents of Lyme disease and relapsing fever, are preferably reduced to zero. Consequently, their toxins are also undetectable. The preferred proportions of sage, ethanol, and honey are approximately 1% honey and 1% extract, where the extract consists of either ethanol and sage or water and sage. The extract preferably consists of a maximum of 10% sage and at least 90% ethanol or water. Ethanol and water can be substituted or combined.

[0095] The sage used can be one or more varieties. A combination of several varieties is therefore possible.

[0096] Patient examples:

[0097] 1.

[0098] A 48-year-old woman was infected multiple times and, despite treatment with espicillin, still developed relapsing-remitting Lyme disease. She then began taking an alcoholic extract (96% ethanol) of Salvia taraxacifolia and Salvia absconditiflora (equal parts) dissolved in honey. She took 2 cl daily, always in the evening. Afterward, she became symptom-free, and no Borrelia bacteria were detectable. Since then, she has occasionally taken the sage extract in honey.

[0099] 2.

[0100] A 36-year-old man noticed a large, circular rash around a tick bite and went to the doctor. The doctor sent him to the hospital where he received an antibiotic infusion. He developed symptoms of Lyme disease. At times, the man could only walk with a walker. The doctor confirmed Lyme disease. The man then started taking an aqueous extract (he cannot tolerate alcohol) of Salvia pisidica and Salvia tomentosa (of which most of the leaves were present) dissolved in honey. He took 2 cl of this extract daily in the evening.

[0101] Starting in the summer, the man noticed an improvement in his symptoms, but no significant improvement. He then added an equal amount of Salvia absconditiflora leaves to the extract. From that point on, his symptoms subsided, and he has remained symptom-free ever since. He has been taking a 20-day course of sage extract once or twice a year, taking 2 cl of the extract in the evening.

[0102] 3.

[0103] A 56-year-old woman had been suffering from multiple sclerosis for years. After lengthy discussions and a review of all her medical records, it turned out that her diagnosis had been incorrect and that she was, in all likelihood, suffering from Lyme disease. Following laboratory testing to confirm this suspicion and the diagnosis of southern Lyme disease, the woman began taking an alcoholic (96%) extract of Salvia absconditiflora and Salvia decumbens (sage chosen for availability) in honey. She took 2 cl of the extract daily in the evening. Subsequently, she became symptom-free, and no Borrelia spp. were detectable.

Claims

Patent claims Claim 1: Ethanolic extract of leaves of one of the Salvia spp. for use in a method for combating borreliosis, comprising the following ingredients: a) Leaves of Salvia spp. selected from the group of sage species consisting of Salvia absconditiflora, Salvia tomentosa, Salvia recognita, Salvia pisidica, Salvia caespitosa, Salvia jurisicii, Salvia nevadensis, Salvia taraxacifolia, Salvia cinerea and / or combinations thereof; b) At least 96% ethanol; and c) Honey. Claim 2: Ethanolic extract for use according to claim 1, wherein the leaves are young. Claim 3: Ethanolic extract for use according to claim 1 or 2, wherein the ethanol is 100% ethanol. Claim 4: Ethanolic extract for use according to one of claims 1 to 3, characterized in that the bee honey has the quality of organic blossom honey and / or rosemary honey. Claim 5: A method for producing a therapeutic composition for combating Lyme disease, comprising the steps of: a) incorporating leaves of Salvia spp. selected from the group of sage species consisting of Salvia absconditiflora, Salvia tomentosa, Salvia recognita, Salvia pisidica, Salvia caespitosa, Salvia jurisicii, Salvia nevadensis, Salvia taraxacifolia, Salvia cinerea and combinations thereof into a quantity of at least 96% ethanol; b) allowing an extract of a) to stand for at least 24 hours at temperatures of about 25 degrees Celsius; c) filtering the leaves; d) incorporating the remaining extract from c) into liquefied blossom honey while stirring constantly. Claim 6: Method according to claim 5, wherein the blossom honey comprises rosemary honey. Claim 7: Method according to at least one of claims 5 or 6, wherein the blossom honey and / or rosemary honey is of organic quality.

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