key: cord-0940943-1pcgw5ey
authors: Carlos Eduardo Cuevas-Barragan; Buenrostro-Nava, Marco Tulio; Palos- Gómez, Gabriela Monserrat; Ramirez-Padilla, Elisa Alejandra; Mendoza-Macias, Blanca Isabel; Rivas-Caceres, Raymundo Rene
title: Use of Nasoill® via intranasal to control the harmful effects of Covid-19
date: 2020-09-17
journal: Microb Pathog
DOI: 10.1016/j.micpath.2020.104504
sha: cb8e5d90eb3a4bdb1b2b1545e20fd4480463657f
doc_id: 940943
cord_uid: 1pcgw5ey

In the absence of vaccines and antiviral drugs available to prevent and treat COVID-19, it becomes imperative to find or use all those products with the potential to fight this virus. This article is an attempt to propose ways to prevent, treat and control the COVID-19 virus, using a product based on plant extracts with the potential to reduce the symptoms caused by the SARS-CoV-2 virus. Nasoil® counts as one of its main components, Asclepias curassavica extracts, and in the present study it has been shown that it is an effective adjuvant in the treatment of Covid-19, increasing the respiratory capacity of the patients (SpO2> 90%) and reducing the symptoms from the first application, improving the patients around the fifth to the eighth application. At a preventive level, the individuals in this study who have applied it (400 individuals) only a 3.15% of these presented symptoms, disappearing when increasing the weekly applications.

have gastrointestinal problems such as nausea, vomiting, diarrhea, abdominal pain, in addition to shortness of breath, headache, sore throat , colds, breathing problems, myalgia, nasal congestion and inflammation of the mucous membranes that cover the inside of the sinuses and nasal discharge, also in some cases occur injuries to the central nervous system [13] [14] [15] [16] [17] [18] [19] [20] .

According to Trisha Greenhalgh et al., (2020) [21] , the best clinical signs for predicting adult acquired pneumonia are a temperature above 38 º C, a breathing rate greater than 20 breaths/minute and a heart rate greater than 100 beats/minute; low urine flow is also a worrying symptom [21] . Two of the main symptoms of Covid-19 include anosmia (loss of smell) and ageusia (loss of taste). Chemosensory dysfunction is necessarily associated with Covid-19 infection and should be considered when detecting symptoms. Understanding the timing and association of odor/taste loss in Covid-19 can help facilitate early detection and isolation of cases. There is a strong association between an olfactory and taste deficiency with a Covid-19 infection. Loss of odors/flavors can be used as a symptom of increased detection of Covid-19 infections to reduce the risk of disease transmission from mildly symptomatic cases [22] .

To date, no adequate vaccine or treatment has been found to combat the pandemic. Some researchers encourage the use of traditional herbal medicine to decrease spread while waiting for the introduction of a vaccine, especially in developing countries; but it must be done according to the guidelines of modern medicine in patient care [23] [24] [25] . Integrating traditional medicine into conventional treatment can be an alternative approach and event to combat resource scarcity.

Clinical studies are needed to evaluate the efficacy and safety of traditional medicines and provide clinical evidence [26] .

Taking into account the above and the wealth of plants in the Mexican Republic, we proceeded to look for plants that could have the best capacity to help control the symptoms caused J o u r n a l P r e -p r o o f by the SARS-CoV-2 virus, in addition to standardizing a protocol to obtain the extracts, having everything ready proceeded to the registration of Nasoil® has different plant extracts, of which one of these extracts are Asclepias curassavica (Apocynaceae) which has different uses, among which they stand out against respiratory, cardiotonic, analgesic and dermatological conditions. The interest in A. curassavica is not only given for its use in traditional Mexican medicine, but even the use of this species in different countries worldwide, highlighting even the name used to a species of this genus in the United States of America which is Pleurisy root, since it allows to cure respiratory conditions, deflating the pleura and lung [27] .

The properties of the species A. curassavica in particular are sneezing, febrifuge and expectorant. Allowing to cure a constipated cold by sneezing the patient, reducing fever and, as we note, reduces inflammation of the pleura (coating of the lungs) due to lung infections hence its name in English [28] . Different studies have shown that extracts obtained through different solvents such as chloroform, alcohols and aqueous A. curassavica presents antimicrobial, antiinflammatory, alexitic, activities, this due to its proteolitic, indirect hemolytic and coagulating capacity. As you can see this plant has a great capacity to modify structures, since it presents some compounds such as cardiotonic glycosides (cardenolides) and various types of alkaloids derived from 2-methoxy-pyrrazine, indole, pyridine or phenantroindolizidine, as well as flavonoids, occasionally tannins, steroids, coumarins, free triterpenes and sesquiterpene lactones [29] [30] [31] [32] [33] . With this context and observing the properties of A. curassavica, which together with the other extracts and molecules have the ability to modify elastic proteins, in addition to helping patients to control the symptoms of the SARS-CoV-2 virus,is that we make available this new product based on extracts from medicinal plants to fight the COVID-19 pandemic.

J o u r n a l P r e -p r o o f

Invasive mechanical ventilation for 10 individuals hospitalized with their respective parameters according to the patient's weight and the degree of loss of respiratory capacity, according to guidelines for patient care by Covid-19, published by the Government of Mexico in February 2020.500 mg azithromycin, broad-spectrum antibiotic for 5 days.500mg Clarithromycin, respiratory tract antibiotic, 500mg paracetamol, analgesic and antipyretic, for fever control every 4 hours, Intramuscular Dexamethasone 8mg, steroid, anti-inflammatory.

Nasoil® is a product with an industrial secret and in patent process that contains extracts of Asclepias currasavica and as well as molecules that help to modify the elastic proteins of the respiratory system and decrease symptoms caused by respiratory diseases.

The protocol for the application of Nasoil ® for patients with symptoms to different degrees of severity without reaching hospitalization was 2 drops of intranasal application per day, i.e. one drop per nostril, for 3 days in a row, and subsequently three applications per week, in case symptoms disappeared earlier or decreased , apps change to 1-2 apps per week, until 20 apps are completed. For patients without symptoms or healthy, the dose was 2 intranasal drops per day per week until 15 applications were completed.

Our study population were 600 individuals dividing them into 6 groups: J o u r n a l P r e -p r o o f GROUP 1: 100 patients confirmed with Covid-19 through laboratory test belonging to the Mexican health sector GROUP 2: it was made up of 100 patients with symptoms (high fever, loss of smell and appetite, diarrhea, phlegm cough) related to SARS-CoV-2 but had no confirmation of positives.

GROUP 3: 99 healthy individuals without symptoms, but at high risk of contagion as they belong to the public health sector. 

The IBM SPSS Statistics 27.0 program was used to perform inter-observer agreement analysis using Cohen's kappa. And the Pearson correlation was performed to determine the degree of relationship between the observed results.

Testing with Nasoil ® began on April 20, 2020, and these observations were completed on July 15, 2020. The effect of Nasoil ® was studied in a total of 600 individuals of which 348 were men and 252 women, the average age was 44.27 years, with ages ranging from 20 to 78 years.

Antibiotics azithromycin were used in 76% of patients confirmed with COVID-19, and in the case of patients with symptoms but without confirmation 10% used this same antibiotic, in addition to this antibiotic, 88 individuals in group 1 used paracetamol 500mg every 4 hours. In group 2, 40% of individuals use this analgesic and antipyretic to control some of the symptoms related to Covid-19, in addition as isolated cases it is worth mentioning, the use of the antibiotic clarithromycin 500mg by an individual in group 1 and another individual in this same group used intramuscular 8mg dexamethasone, the rest of the individuals (472) included part of group 2 and all individuals from groups 3 to 6 , they did not take medications to control symptoms related to Covid-19.

The 90% of patients in group 1 and 2 had the following symptoms: fever, headache, cough, asthenia, anosmia and ageusia with different degree of severity, 10% of patients had only anosmia and ageusia, such loss of smell did not link it to SARS-CoV-2 since there was no presence of any other symptoms (Table 1) .

When the drops were placed in the nostrils, immediately all individuals manifested one or some of the reactions shown in Table 2 . Other reactions were also present which they were discarded, these were the presence of febriculas, diarrhea, low blood sugar levels which were observed mainly in patients confirmed to Covid-19 and sporadically in unconfirmed patients and being null for groups with individuals without symptoms.

From these reactions, it was determined whether they were unique to the application of Nasoil ® droplets or was a reaction as a result of the disease or respiratory discomfort suffered by individuals. Reactions occur in a large number of individuals, such as burning, tearing, nasal flow, and coughing in all groups. Cohen's Kappa was carried out to determine that data obtained by two groups of observers (homeopathic physicians and researchers) were not randomly ob-J o u r n a l P r e -p r o o f tained data, and the Landis and Koch scale (1977) was considered, which is frequently used to qualitatively express the force of concordance between observers (Table 3) .

According to Landis and Koch, (1977) [34] the Kappa concordance coefficient of our two observer groups for each study group is almost perfect, as it is in that range according to Table 3 , it should be clarified that in this study each group of observers checked separately approximately 50 different individuals for each group, subsequently the information was collected in a total of all observed individuals , using the value 1 for those who had had a reaction x and zero for those responses that did not include such a reaction, obtaining as a first impression the percentages shown in Table 2 .

The observations made by the two research groups are consistent that Nasoil ® provokes reactions in all study groups, being almost perfect for groups 1 and 2 (Table 4 ) which are the groups where individuals with symptoms related to Covid-19 are concentrated, having the lower values for Kappa, for the group of individuals considered high risk (group 4) working in the health sector, and the low-risk group considered to be healthy individuals (group 6), all values are in the near-perfect range according to Table 3 .

In Table 5 shows the obtained values for each reaction per study group, having values in the ranges of almost perfect concordance in most groups for each reaction, having values of considerable in the tearing reaction and headache for group 5 and moderate in headache for group 6.

Each observer team was composed, the first, by 3 homeopathic physicians, and the second by ple between the ages of 20 and 30, predominating 60% in women, these individuals noted that immediately after the burning that occurs by the placement of Nasoil ® , they felt the nostrils decongestant and immediately afterwards they could breathe better and begin to perceive the smells slightly.

The 100% of individuals with both confirmed and unconfirmed Covid-19 symptoms initiated the placement of Nasoil ® drops between 4 to 17 days after the first discomfort began. 12 of the individuals initiated the drops hospitalized or when leaving the hospital. In Figure 2 the percentage of all individuals with symptoms related to Covid-19 (Groups 1 and 2 of study) and their status during this study is observed. Of the number of hospitalized (12 individuals), 10 individuals (7 men and 3 women) were already in that state at the beginning of the present work, and a couple was hospitalized after the use of the droplets. In the case of the 10 hospitalized individuals, they were with invasive mechanical ventilation, of which 8 (5 men and 3 women) were applied Nasolil ® during their hospitalization and 2 (both men) began post-hospitalization application.

The 8 individuals who were applied to Nasoil ® in the hospital, had acute respiratory distress syndrome (ARDS) in addition to having onset of chronic obstructive pulmonary disease (COPD), having SpO 2 values less than 40%. At the time of use of Nasoil ® all 8 patients no longer required mechanical ventilation from the first intake, and even one of the 3 women referred that ventilation will be removed at the minute of the application of the droplets, all patients were discharged without any complication, having SpO2 values greater than 90%.

Of the 2 patients who were with invasive mechanical assistance, but were not given the drops during their hospitalization but after it, they spent 17 days in the hospital, being 2 male individuals aged 38 and 57, the individual aged 38, has no harm, however he was discharged with oxygen assistance, and recurrent extreme weakness and dry cough that prevented rest , at the time of the J o u r n a l P r e -p r o o f use of Nasoil ® the help of oxygen by mask, flung phlegms with abundant nasal flow was withdrawn, which allowed him to improve his breathing and the recurrent dry cough disappeared.

The patient aged 57 years was discharged with a pulmonary microthrobosis and cerebral hypoxia, maintaining SpO 2 values less than 90%, extreme tiredness that prevented him from walking and even sitting in the bed itself, at the time of use of Nasoil ® did not require the help of oxygen by mask, his oxygenation in blood increased to 96%, tiredness and constant cough remitted , it should be mentioned that this individual has started to take his first steps with help, will prolong the application of Nasoil ® hopefully improve even more.

Two individuals who were given the drops after hospitalization, are a couple with ages 64 for the woman and 67 years the man, the first to be hospitalized was the man, he applied the drops, only once four days before being hospitalized, the reason for the application was the presence of flu symptoms, he stopped using them , and was subsequently hospitalized, in his hospitalization did not require invasive mechanical ventilation, and during the 14 days of his hospital was given Nasoil ® only one occasion, at the time of being transferred to another hospital confirming that he had Covid-19, and due to the protocols available to the public health sector could not be applied the drops, at the time of stopping using them, his SpO 2 under 70%, however it did not require assisted ventilation, once outside the hospital, returned to the application of the drops, maintaining a SpO 2 between 92 and 96%, expulsion of phlegms and abundant nasal flow, improving its breathing capacity, and completely eliminated fever and general discomfort. The woman at the time of hospitalization her husband, start with azithromycin treatment for 5 days, and then start with the application of Nasoil ® , was hospitalized 4 days after Nasoil ® use, she removed the application of the drops, lowering her SpO 2 , to 80%, however did not require invasive ventilation, J o u r n a l P r e -p r o o f lasting 7 days in the hospital, restarting Nasoil ® applications when finished her boarding hospital and after the 14th application of Nasoil ® was found discharged.

The rest of the individuals positive to Covid-19 had improvements in some cases almost immediately, 60% of the individuals mentioned that they felt improvement from the first application and even though the fever in almost 80% of them remained, all noting that tiredness and shortness of breath had disappeared.

Group 2 all improved, immediately, and those who had severe symptoms felt their improvement between the 5 and 8 application. Groups 3 to 6, regardless of the 13 individuals who indicated that they started with the presence of symptoms (Figure 1 ), are all found without the presence of symptoms or discomfort. The reactions caused by Nasoil ® in all individuals in which they occurred lasted no longer than a few seconds in the case of burning and tearing, a couple of minutes for nasal flow, sneeze in a burst and cough, and for the discomfort of the head either pain or dizziness, about 5 to 10 min.

Dizziness occurred consistently in individuals who claimed to have recurrent hearing infections, or even those who indicated a problem at the nasal septum level. However, this discomfort was not prolonged and in 100% of cases was not an impediment to pursuing its tasks.

Nasoil ® has as one of its main components extracts of Asclepias curassavica and was mainly intended to improve the elastic capacity of the lung, since information is reported on its ability to modify proteins [35], to activate change at the cellular level [36,37], act as protease causing protein degradation due to modification of intramolecular changes [35-38] with these characteristics is intended to resolve some respiratory diseases such as pulmonary emphysema, due to the cur-J o u r n a l P r e -p r o o f rent contingency he proceeded to study whether he could be an assistant to reduce the number of deaths by Covid-19, and above all as a preventative, by improving breathing capacity, a fact seen in hospitalized patients who went from having SpO 2 values < 40% and when using the drops the values were SpO 2 > 90%.

Of the main symptoms seen in Table 1 that individuals infected with the SARS-CoV-2 virus improve from the first application is their ability to breathe, as mentioned above, this improvement in breathing is accompanied by an improvement in asthenia, anosmia and eugesia, the next symptom that improves with applications is fever which although present ceases to be high , the decrease in these malaises present by Covid-19 may be due to the addition of Asclepias curassavica extracts has been shown in some studies to have their analgesic, antipyretic, and antiinflammatory capacity [39,40].

The reason for the use of Nasoil ® via intranasal administration is in order to affect steroid receptors, as it has been shown that, by this means, there is a greater benefit in neuroinflamation- In the individuals patients who lasted 17 days hospitalized and discharged, had severe damage at the lung level and even one was discharged with a pulmonary microthrombosis and cerebral hypoxia, at the time of the use of Nasoil ® started with abundant nasal flow and removal of phlegms by cough, this patient, like the patient who also applied the drops after their hospitalization had a cough that did not allow them to sleep , and with the lung problems present maintained a state of severe fatigue, as Nasoil applications progressed ® , both had decreased cough and ability to breathe improvement. It is clear that Nasoil ® produced anti-inflammation, and decreases the microthrombosis caused by hypercoagulation due to polymerization and fibrin formation [47], it is inferred that this last effect on coagulation is given to the characteristic observed in Asclepias extracts decrease coagulation caused by some poisons, through the cardiotonic glycosides present in this plant [48], these two actions, anti -inflammation and decreased clotting, help to have a better ability to breathe. Nasoil ® emerges as a product with the ability to modify the elastic proteins of the respiratory system, mainly to modify elastin, since if this elastic protein is damaged, due to inflammation, pres- 

On the other hand, the elastic fibres around the more distal airways and alveolar sacs change in their location and orientation and become thinner [50] . Therefore, reinforcing elastin or stimulating other critical residues that assume a new role and allow it to form new interactions that would prevent its degradation or structural modification, which would prevent the loss of lung activity. The individuals in group 5 were made up of individuals who presented other respiratory problems, such as bronchitis, asthma, allergies, sinusitis, and even patients who did not have respiratory diseases, but were smokers, 100% presented improvement in their conditions and even smokers showed an improvement in their breathing which has manifested itself with improvement at bedtime. In the same way within the other groups, there were people with some similar problems to those of this particular group, and in the same way they presented improvement to the use of Nasoil ® .

Anfinsen points out that the three-dimensional structure of a protein is determined by the sequence of amino acids. This dogma has been a central principle of molecular biology and biochemistry, however, it is not entirely true since the inability of x-ray crystallography to resolve the structures of many proteins and protein segments, often, Anfinsen's dogma applies only to the domains of a protein since proteins can also exist in inherently disordered states and cannot be described by a single atomic resolution conformation.

The relative disposition of the domain may change through large-scale conformational transitions following interaction with small ligands, other proteins, nucleic acids and even microenvironmental changes such as pH, temperature etc., a review of Anfinsen's dogma is necessary [51, 52] . Clarke (2020) points out that, the sequence of amino acids of a protein must encode both order and disorder, or perhaps complete that the protein sequence allows changes depending on [53], the presence of critical residues and a specific microenvironment is necessary for the formation of bonds in proteins, but these bonds can be modified by modifying the environment.

In Morales´ (2010) work with wheat proteins, mainly gluten proteins, which are elastic proteins, he observed that using different substances he had the capacity to generate diverse links in the proteins [56] . Nasoil ® being a combination of plant extracts that have the capacity to modify the tissue microenvironment by oxidizing or reducing depending on the state in which the proteins are found, is that they provide immediately the improvement in the respiratory capacity.

If the elastic proteins are compromised it is necessary that they are modified to retake their capacity, this of indirect form is observed, since 95% of the individuals with confirmation of COVID-19 or associated symptoms without confirmation manifest at the moment of the placement of the drops the imperative need to cough, being with phlegm in 90% of the cases, with nasal flow in 80% and sneeze in 80% of the cases and these manifestations allowed them once they passed, in questions of minutes, a generalized improvement. In addition, 50% of the individuals J o u r n a l P r e -p r o o f confirmed with SARS-CoV-2 when performing new PCR test are negative, this indicates that the structural modification probably helps to prevent the replication of the virus.

The other indirect observation of the change at a structural level is in the individual with pulmonary microthrombosis, who after the applications of Nasoil ® can breathe without external help, has been able to leave the extreme fatigue that prevented him from moving, in this case we are not seeing a preventive aspect, but a reversion to the structural damage of the lung.

Out of a population of 600 individuals analyzed, 100% show an improvement in breathing, including individuals belonging to groups 3 to 6 considered preventive or healthy, most of them showing blood oxygen values higher than 96%. Nasoil ® presents a way of action for the control of the symptoms present in the infected individuals with the SARS-CoV-2 virus and is the one of modifying the pulmonary microenvironment, promoting a change in the microstructure of the elastic proteins, which allows to reduce the inflammation and the coagulation, improving with it the capacity of the individuals to breathe.

As a preventive, Nasoil ® presents a high percentage of protection to the presence of symptoms (~96%), this does not mean that the individuals do not get infected, but that they do not show in an aggressive way the problems caused by the SARS-CoV-2 virus, and even allowing an immediate improvement in case these symptoms appear.

Nasoil ® does not affect the SARS-CoV-2 virus but its action is to improve the capacity of the lungs, improving their elastic capacity and preventing the possible hypercoagulation, and the ex-J o u r n a l P r e -p r o o f aggerated action of the immune system of the individuals (neutrophilia), by stimulating the elastic proteins.

In individuals with severe Covid-19 disease, Nasoil ® acts as a stimulant of the action to eliminate the phlegm caused by the lung inflammation, and as a mechanism to expel the virus by mechanical action, such as coughing, nasal flow and even diarrhea in some individuals.

Of the total number of confirmed with Covid-19 all improved and 50% of them were negative when new PCR test. Currently in Mexico, homeopathic physican are using it as a coadjuvant to treat the symptoms of Covid-19 and other respiratory diseases.

Much work remains to be done to better determine the ways in which Nasoil ® acts on the body to improve its condition, as well as to determine the actions on other respiratory diseases, which were seen during the performance of this work. Nasoil ® is not a cure or a vaccine, is an adjuvant for the control of symptoms and discomfort caused by the SARS-CoV-2 virus. J o u r n a l P r e -p r o o f J o u r n a l P r e -p r o o f J o u r n a l P r e -p r o o f 

Now casting and forecasting the potential domestic and international spread of the 2019-nCoV outbreak originating in Wuhan, China: a modelling study

Outbreak of pneumonia of unknown etiology in Wuhan China: the mystery and the miracle

Histopathologic changes and SARS-CoV-2 immunostaining in the lung of a patient with COVID-19

Aerosol and surface stability of SARS-CoV-2 as compared with SARS-CoV-1

Clinical characteristics of 138 hospitalized patients with 2019 novel coronavirus-infected pneumonia in Wuhan

Risk factors associated with acute respiratory distress syndrome and death in patients with coronavirus disease 2019 pneumonia in Wuhan, China

Study of combining virtual screening and antiviral treatments of Sars-CoV-2 (Covid-19)

Lung recruitability in SARS-CoV-2 associated acute respiratory distress syndrome: A single-center, observational study

Clinical features of patients infected with 2019 novel coronavirus in Wuhan

Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study

Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study

Clinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected: interim guidance

neutrophil-to-lymphocyte ratio and IgG predicts disease severity and outcome for patients with COVID-19. med Rxiv

Correlation analysis between disease severity and inflammation-related parameters in patients with COVID-19 pneumonia

Covid-19: a remote assessment in primary care

COVID-19): A protocol for prevention and treatment (Covalyse ® )

Network pharmacology-based analysis of the role of traditional Chinese herbal medicines in the treatment of COVID-19

Will plant-made biopharmaceuticals play a role in the fight against COVID-19?

Accelerated production of hesperidin-rich citrus pectin from waste citrus peel for prevention and therapy of COVID-19

Usos de las especies del género Asclepias L. (APOCYNACEAE, ASCLEPIADOIDEAE), información del herbario nacional de México

Neutrophil extracellular traps (NETs) as markers of disease severity in COVID-19

Neutrophil extracellular traps kill bacteria

Excessive neutrophils and neutrophil extracellular traps contribute to acute lung injury of influenza pneumonitis

Extracellular histones are major mediators of death in sepsis